cytokine  (Thermo Fisher)


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    eBioscience Mouse Cytokine Positive Control Cells
    Description:
    The Mouse Cytokine Positive Control Cells are recommended for use as positive control cells when establishing an intracellular cytokine staining protocol Mouse splenocytes were stimulated with ConA 3 µg mL for two days followed by recombinant IL 2 20 ng mL and IL 4 20 ng mL for three days The cells were then restimulated with immobilized anti mouse CD3 10 µg mL and soluble anti mouse CD28 2 µg mL in the presence of Brefeldin A for 5 hours These cells contain subpopulations that express IL 2 IL 3 IL 4 IL 10 GM CSF and IFNγ The cells have been fixed with formaldehyde and are provided in a stabilizing buffer so that they may be stored at 80°C for up to six months or at 2 8°C for up to one month Reactivity SpeciesMouseReported ApplicationIntracellular Staining Followed by Flow Cytometric Analysis
    Catalog Number:
    00-4500-51
    Price:
    None
    Category:
    Lab Reagents and Chemicals
    Applications:
    Cell Analysis|Cellular Imaging|Flow Cytometry
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    Structured Review

    Thermo Fisher cytokine
    Expansion of Th17 cells in PBMCs of patients with active SSc. (A) Human PBMCs were labeled with antibody against lymphocytes (anti-CD3 and -CD8). IL-17-expressing cells were detected by intracellular <t>cytokine</t> staining assay, and the percentage of IL-17 + cells among CD3 + CD8 - T cells was determined with flow cytometry. (B) Results of flow-cytometric analysis of Th17 cells in patients with active SSc ( n = 13), patients with stable SSc ( n = 32), and controls ( n = 24). (C) Longitudinal monitoring of Th17 cells in 10 patients. The percentage of Th17 cells was measured initially during active SSc and again after resolution after treatment. (D) Real-time RT-PCR analysis of IL-17, Foxp3, and RORγt mRNA expression in freshly isolated PBMCs of patients with active SSc ( n = 13), patients with stable SSc (n = 32), and controls (n = 24). (E) Positive correlation between the proportion of Th17 cells and clinical severity in active SSc patients, scored by using the Valentini score ( n = 13). r = 0.675, P
    The Mouse Cytokine Positive Control Cells are recommended for use as positive control cells when establishing an intracellular cytokine staining protocol Mouse splenocytes were stimulated with ConA 3 µg mL for two days followed by recombinant IL 2 20 ng mL and IL 4 20 ng mL for three days The cells were then restimulated with immobilized anti mouse CD3 10 µg mL and soluble anti mouse CD28 2 µg mL in the presence of Brefeldin A for 5 hours These cells contain subpopulations that express IL 2 IL 3 IL 4 IL 10 GM CSF and IFNγ The cells have been fixed with formaldehyde and are provided in a stabilizing buffer so that they may be stored at 80°C for up to six months or at 2 8°C for up to one month Reactivity SpeciesMouseReported ApplicationIntracellular Staining Followed by Flow Cytometric Analysis
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    1) Product Images from "Increased frequency of Th17 cells in systemic sclerosis is related to disease activity and collagen overproduction"

    Article Title: Increased frequency of Th17 cells in systemic sclerosis is related to disease activity and collagen overproduction

    Journal: Arthritis Research & Therapy

    doi: 10.1186/ar4430

    Expansion of Th17 cells in PBMCs of patients with active SSc. (A) Human PBMCs were labeled with antibody against lymphocytes (anti-CD3 and -CD8). IL-17-expressing cells were detected by intracellular cytokine staining assay, and the percentage of IL-17 + cells among CD3 + CD8 - T cells was determined with flow cytometry. (B) Results of flow-cytometric analysis of Th17 cells in patients with active SSc ( n = 13), patients with stable SSc ( n = 32), and controls ( n = 24). (C) Longitudinal monitoring of Th17 cells in 10 patients. The percentage of Th17 cells was measured initially during active SSc and again after resolution after treatment. (D) Real-time RT-PCR analysis of IL-17, Foxp3, and RORγt mRNA expression in freshly isolated PBMCs of patients with active SSc ( n = 13), patients with stable SSc (n = 32), and controls (n = 24). (E) Positive correlation between the proportion of Th17 cells and clinical severity in active SSc patients, scored by using the Valentini score ( n = 13). r = 0.675, P
    Figure Legend Snippet: Expansion of Th17 cells in PBMCs of patients with active SSc. (A) Human PBMCs were labeled with antibody against lymphocytes (anti-CD3 and -CD8). IL-17-expressing cells were detected by intracellular cytokine staining assay, and the percentage of IL-17 + cells among CD3 + CD8 - T cells was determined with flow cytometry. (B) Results of flow-cytometric analysis of Th17 cells in patients with active SSc ( n = 13), patients with stable SSc ( n = 32), and controls ( n = 24). (C) Longitudinal monitoring of Th17 cells in 10 patients. The percentage of Th17 cells was measured initially during active SSc and again after resolution after treatment. (D) Real-time RT-PCR analysis of IL-17, Foxp3, and RORγt mRNA expression in freshly isolated PBMCs of patients with active SSc ( n = 13), patients with stable SSc (n = 32), and controls (n = 24). (E) Positive correlation between the proportion of Th17 cells and clinical severity in active SSc patients, scored by using the Valentini score ( n = 13). r = 0.675, P

    Techniques Used: Labeling, Expressing, Staining, Flow Cytometry, Cytometry, Quantitative RT-PCR, Isolation

    2) Product Images from "Th17 cells favor inflammatory responses while inhibiting type I collagen deposition by dermal fibroblasts: differential effects in healthy and systemic sclerosis fibroblasts"

    Article Title: Th17 cells favor inflammatory responses while inhibiting type I collagen deposition by dermal fibroblasts: differential effects in healthy and systemic sclerosis fibroblasts

    Journal: Arthritis Research & Therapy

    doi: 10.1186/ar4334

    Th17 cell clone generation and characterization. (A , B) Boolean gating analysis showing all combinations of IL-17A, IL-22 and IFN-γ production by CD4 + CD45RA- memory T cells before and after stepwise enrichment based on CCR6 + CCR4 + CCR10- and CD161+ surface expression (A) , and in the 20 T cell clones expanded after enrichment (B) . Intracellular cytokine staining was performed after PMA and ionomycin stimulation. Color codes are shown in the right panel of A. The green circular segment indicates the total number of IL-17A-producing T cells after each step of enrichment. (C) T cell cytokine levels were measured in supernatants of five of the twenty activated Th17 cell clones by ELISA and bead immunoassay. CCR: CC-chemokine receptor; CD: cluster of differentiation; ELISA: enzyme immunosorbent assay; IFN: interferon; IL: interleukin; PMA: phorbol myristate acetate.
    Figure Legend Snippet: Th17 cell clone generation and characterization. (A , B) Boolean gating analysis showing all combinations of IL-17A, IL-22 and IFN-γ production by CD4 + CD45RA- memory T cells before and after stepwise enrichment based on CCR6 + CCR4 + CCR10- and CD161+ surface expression (A) , and in the 20 T cell clones expanded after enrichment (B) . Intracellular cytokine staining was performed after PMA and ionomycin stimulation. Color codes are shown in the right panel of A. The green circular segment indicates the total number of IL-17A-producing T cells after each step of enrichment. (C) T cell cytokine levels were measured in supernatants of five of the twenty activated Th17 cell clones by ELISA and bead immunoassay. CCR: CC-chemokine receptor; CD: cluster of differentiation; ELISA: enzyme immunosorbent assay; IFN: interferon; IL: interleukin; PMA: phorbol myristate acetate.

    Techniques Used: Expressing, Clone Assay, Staining, Enzyme-linked Immunosorbent Assay

    3) Product Images from "CCR4 Participation in Th Type 1 (Mycobacterial) and Th Type 2 (Schistosomal) Anamnestic Pulmonary Granulomatous Responses 1"

    Article Title: CCR4 Participation in Th Type 1 (Mycobacterial) and Th Type 2 (Schistosomal) Anamnestic Pulmonary Granulomatous Responses 1

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi:

    Sensitized wild-type CCR4 +/+ CD4 + T cells display diminished IFN-γ production when cultured with CCR4 −/− DCs. CD4 + T cells and CD11c + DCs were isolated from the axillary lymph nodes of PPD-sensitized donors. CCR4 +/+ CD4 + T cells were cultured with either CCR4 +/+ or CCR4 −/− DCs at a ratio of 10:1 for 72 h with PPD Ag. Cytokine levels were determined in culture supernatants by ELISA. Data are representative of two separate experiments, four to five mice per group. Bars are means ± SD. *, p
    Figure Legend Snippet: Sensitized wild-type CCR4 +/+ CD4 + T cells display diminished IFN-γ production when cultured with CCR4 −/− DCs. CD4 + T cells and CD11c + DCs were isolated from the axillary lymph nodes of PPD-sensitized donors. CCR4 +/+ CD4 + T cells were cultured with either CCR4 +/+ or CCR4 −/− DCs at a ratio of 10:1 for 72 h with PPD Ag. Cytokine levels were determined in culture supernatants by ELISA. Data are representative of two separate experiments, four to five mice per group. Bars are means ± SD. *, p

    Techniques Used: Cell Culture, Isolation, Enzyme-linked Immunosorbent Assay, Mouse Assay

    Purification of IFN-γ and IL-4 cytokine-producing CD4 + T cells from lungs during type-1 (PPD) and type-2 (SEA) granuloma formation. Anamnestic lung granulomas were induced in CBA/J mice and on day 3, lungs were collected, homogenized, and cultured overnight with Ag. CD4 + cytokine-secreting populations were isolated as described in Materials and Methods. Relative mRNA transcript levels were measured for IFN-γ ( A ) and IL-4 ( B ) with results expressed as the fold increase over the nonenriched CD4 + control (ALL). Insets , The expression in arbitrary units of the CD4 + nonenriched control demonstrates the initial cytokine polarization. Transcript levels of chemokine receptors were measured by real-time RT-PCR in the enriched populations. C , CCR4 transcripts. D , CXCR3 transcripts. Type-1 (PPD) response,█; type-2 (SEA) response,▒. Bars are mean arbitrary units ± SD derived from two separate experiments, five mice per experiment.
    Figure Legend Snippet: Purification of IFN-γ and IL-4 cytokine-producing CD4 + T cells from lungs during type-1 (PPD) and type-2 (SEA) granuloma formation. Anamnestic lung granulomas were induced in CBA/J mice and on day 3, lungs were collected, homogenized, and cultured overnight with Ag. CD4 + cytokine-secreting populations were isolated as described in Materials and Methods. Relative mRNA transcript levels were measured for IFN-γ ( A ) and IL-4 ( B ) with results expressed as the fold increase over the nonenriched CD4 + control (ALL). Insets , The expression in arbitrary units of the CD4 + nonenriched control demonstrates the initial cytokine polarization. Transcript levels of chemokine receptors were measured by real-time RT-PCR in the enriched populations. C , CCR4 transcripts. D , CXCR3 transcripts. Type-1 (PPD) response,█; type-2 (SEA) response,▒. Bars are mean arbitrary units ± SD derived from two separate experiments, five mice per experiment.

    Techniques Used: Purification, Crocin Bleaching Assay, Mouse Assay, Cell Culture, Isolation, Expressing, Quantitative RT-PCR, Derivative Assay

    4) Product Images from "Live Attenuated Leishmania donovani Centrin Knock Out Parasites Generate Non-inferior Protective Immune Response in Aged Mice against Visceral Leishmaniasis"

    Article Title: Live Attenuated Leishmania donovani Centrin Knock Out Parasites Generate Non-inferior Protective Immune Response in Aged Mice against Visceral Leishmaniasis

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0004963

    Leishmania Ag–stimulated cytokine profiles in splenocytes culture supernatants from naive, LdCen-/- immunized (Imm), naive challenged (Naive Chal), and LdCen-/- immunized challenged (Imm Chal) young and aged mice. The 5-wk post immunized mice were challenged with virulent L . donovani for 4-wk and then mice were euthanized, splenocytes were isolated, plated aseptically (2×10 5 cells/well), and stimulated with Leishmania FTAg for 72h. Concentrations of pro-inflammatory cytokines IFN-γ (A), IL-12 (B) and TNF (C) anti-inflammatory cytokines IL-10 (D) and IL-4 (E) were measured in culture supernatants using the multiplex mouse cytokine kit as described in the Material and Methods section. Ratio of IFN-γ/IL-10 (F) and IFN-γ/IL-4 (G) were also determined. The data presented are representative of two independent experiments with similar results (n = 6). Mean and SEM of each group are shown. * p
    Figure Legend Snippet: Leishmania Ag–stimulated cytokine profiles in splenocytes culture supernatants from naive, LdCen-/- immunized (Imm), naive challenged (Naive Chal), and LdCen-/- immunized challenged (Imm Chal) young and aged mice. The 5-wk post immunized mice were challenged with virulent L . donovani for 4-wk and then mice were euthanized, splenocytes were isolated, plated aseptically (2×10 5 cells/well), and stimulated with Leishmania FTAg for 72h. Concentrations of pro-inflammatory cytokines IFN-γ (A), IL-12 (B) and TNF (C) anti-inflammatory cytokines IL-10 (D) and IL-4 (E) were measured in culture supernatants using the multiplex mouse cytokine kit as described in the Material and Methods section. Ratio of IFN-γ/IL-10 (F) and IFN-γ/IL-4 (G) were also determined. The data presented are representative of two independent experiments with similar results (n = 6). Mean and SEM of each group are shown. * p

    Techniques Used: Mouse Assay, Isolation, Multiplex Assay

    Ag-specific intracellular cytokine secretion analysis of CD4 and CD8 T cells from LdCen-/- immunized and non-immunized young and aged mice after virulent L . donovani challenge. The 8- wk post-immunized or non-immunized young and aged mice were challenged for 4-wk with virulent L . donovani . Intracellular cytokine analysis was done as shown in Fig 6A and divided into seven distinct subpopulations. (A) Cytokine analysis of CD4 T cells from 8-wk post immunized and 4-wk post challenged mice. (B) Cytokine analysis of CD8 T cells of 8-wk post immunized and 4-wk post challenged mice. (C) IL-10 secreting CD4 T cells and (D) the ratio of IFN-γ to IL-10 producing CD4 T cells from spleens at the time of challenge [(naive and immunized (8W)] and after challenge [(naive-challenged and immune-challenged (8WI plus 4WPC)]. The data presented are representative of two experiments with similar results. Mean and SEM of six mice in each group are shown. * p
    Figure Legend Snippet: Ag-specific intracellular cytokine secretion analysis of CD4 and CD8 T cells from LdCen-/- immunized and non-immunized young and aged mice after virulent L . donovani challenge. The 8- wk post-immunized or non-immunized young and aged mice were challenged for 4-wk with virulent L . donovani . Intracellular cytokine analysis was done as shown in Fig 6A and divided into seven distinct subpopulations. (A) Cytokine analysis of CD4 T cells from 8-wk post immunized and 4-wk post challenged mice. (B) Cytokine analysis of CD8 T cells of 8-wk post immunized and 4-wk post challenged mice. (C) IL-10 secreting CD4 T cells and (D) the ratio of IFN-γ to IL-10 producing CD4 T cells from spleens at the time of challenge [(naive and immunized (8W)] and after challenge [(naive-challenged and immune-challenged (8WI plus 4WPC)]. The data presented are representative of two experiments with similar results. Mean and SEM of six mice in each group are shown. * p

    Techniques Used: Mouse Assay

    Multiparameter flow cytometry based analysis for single, double, or triple cytokine–secreting CD44 Hi /CCR7 Low / CD4 or CD8 T cells from LdCen-/- immunized and non-immunized young and aged mice. (A) The common gating steps shown in this study. Spleen cells of 8-wk post immunized mice were stimulated with FTAg for 48h and stained with various Abs as described in Materials and Methods. Ag-experienced effector cells were gated and divided into seven distinct subpopulations, and the frequencies of the various subpopulations were calculated. (B) Cytokine analysis of CD4 T cells from naïve and 8-wk post-immunized mice. ( C ) Cytokine analysis of CD8 T cells from naïve and 8-wk post-immunized mice. The data presented are representative of two experiments with similar results. Mean and SEM of six in each group is shown. * p
    Figure Legend Snippet: Multiparameter flow cytometry based analysis for single, double, or triple cytokine–secreting CD44 Hi /CCR7 Low / CD4 or CD8 T cells from LdCen-/- immunized and non-immunized young and aged mice. (A) The common gating steps shown in this study. Spleen cells of 8-wk post immunized mice were stimulated with FTAg for 48h and stained with various Abs as described in Materials and Methods. Ag-experienced effector cells were gated and divided into seven distinct subpopulations, and the frequencies of the various subpopulations were calculated. (B) Cytokine analysis of CD4 T cells from naïve and 8-wk post-immunized mice. ( C ) Cytokine analysis of CD8 T cells from naïve and 8-wk post-immunized mice. The data presented are representative of two experiments with similar results. Mean and SEM of six in each group is shown. * p

    Techniques Used: Flow Cytometry, Cytometry, Mouse Assay, Staining

    Analysis of young and aged mice derived dendritic cell function upon LdWT and LdCen-/- infection in vitro . BMDCs isolated from young and aged groups of mice were infected with LdWT or LdCen-/- stationary-phase promastigotes (5:1, parasite to DCs ratio) and intracellular parasite numbers were visualized by Giemsa staining and estimated microscopically at 6, 24, 48 and 72h post-infection. (A) Infection efficiency (% of infected cells) and (B) intracellular growth (parasites per infected cell) were recorded. To measure parasite load in these cultures, a minimum of 300 BMDCs were counted. In a separate experiment BMDCs were infected with parasites and stimulated with LPS (1 μg/ml) for 24h. Culture supernatants were collected to analyze IL-12 (C), TNF (D) IL-6 (E) production by ELISA and NO (F) production by the Griess assay. The data presented are representative of two independent experiments. T cell proliferation and cytokine production upon coculture of parasite-infected BMDCs with OVA-specific transgenic T cells. BMDCs obtained from young and aged BALB/c mice were pulsed with OVA peptide and were either left uninfected or infected with LdWT or LdCen-/- for 24h. CD4 + T cells were purified from age matched young and aged DO11.10 transgenic mice, stained with CFSE and co-cultured with parasite infected BMDCs for 5 days. (G) T cell proliferation was estimated by flow cytometry by studying CFSE dilution of gated CD4 + T cells and represented by the histogram. The staggered histogram overlay displays CD4 + T cell proliferation pattern as visualized by CFSE dilution by flow cytometry. Cell proliferation was done in triplicates and histograms representative of mean values were overlaid for figure. The black line on histogram over lay represents % proliferated cells gated in CD4 + T lymphocytes. (H) The bar diagram representing the quantitative CFSE cell proliferation. (I, J) Culture supernatants were collected at day 5 of coculture to assay IFN-γ and IL-10 by ELISA. (K) IFN-γ: IL-10 ratio was determined. The data represent the mean values ± SD of results from 3 independent experiments that all yielded similar results. * p
    Figure Legend Snippet: Analysis of young and aged mice derived dendritic cell function upon LdWT and LdCen-/- infection in vitro . BMDCs isolated from young and aged groups of mice were infected with LdWT or LdCen-/- stationary-phase promastigotes (5:1, parasite to DCs ratio) and intracellular parasite numbers were visualized by Giemsa staining and estimated microscopically at 6, 24, 48 and 72h post-infection. (A) Infection efficiency (% of infected cells) and (B) intracellular growth (parasites per infected cell) were recorded. To measure parasite load in these cultures, a minimum of 300 BMDCs were counted. In a separate experiment BMDCs were infected with parasites and stimulated with LPS (1 μg/ml) for 24h. Culture supernatants were collected to analyze IL-12 (C), TNF (D) IL-6 (E) production by ELISA and NO (F) production by the Griess assay. The data presented are representative of two independent experiments. T cell proliferation and cytokine production upon coculture of parasite-infected BMDCs with OVA-specific transgenic T cells. BMDCs obtained from young and aged BALB/c mice were pulsed with OVA peptide and were either left uninfected or infected with LdWT or LdCen-/- for 24h. CD4 + T cells were purified from age matched young and aged DO11.10 transgenic mice, stained with CFSE and co-cultured with parasite infected BMDCs for 5 days. (G) T cell proliferation was estimated by flow cytometry by studying CFSE dilution of gated CD4 + T cells and represented by the histogram. The staggered histogram overlay displays CD4 + T cell proliferation pattern as visualized by CFSE dilution by flow cytometry. Cell proliferation was done in triplicates and histograms representative of mean values were overlaid for figure. The black line on histogram over lay represents % proliferated cells gated in CD4 + T lymphocytes. (H) The bar diagram representing the quantitative CFSE cell proliferation. (I, J) Culture supernatants were collected at day 5 of coculture to assay IFN-γ and IL-10 by ELISA. (K) IFN-γ: IL-10 ratio was determined. The data represent the mean values ± SD of results from 3 independent experiments that all yielded similar results. * p

    Techniques Used: Mouse Assay, Derivative Assay, Cell Function Assay, Infection, In Vitro, Isolation, Staining, Enzyme-linked Immunosorbent Assay, Griess Assay, Transgenic Assay, Purification, Cell Culture, Flow Cytometry, Cytometry

    5) Product Images from "IL-1α Signaling Is Critical for Leukocyte Recruitment after Pulmonary Aspergillus fumigatus ChallengeCompartment-specific and sequential role of MyD88 and CARD9 in chemokine induction and innate defense during respiratory fungal infection"

    Article Title: IL-1α Signaling Is Critical for Leukocyte Recruitment after Pulmonary Aspergillus fumigatus ChallengeCompartment-specific and sequential role of MyD88 and CARD9 in chemokine induction and innate defense during respiratory fungal infection

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1004625

    IL-1α signaling enhances expression of leukocyte recruiting chemokines. C57BL/6 mice treated with either isotype control antibody or IL-1α neutralizing antibody, Il1r1 -deficient and Pycard -deficient mice were infected with 5×10 7 CEA10 conidia and at 24 hours post-infection, mice were euthanized, BALF collected, and lung tissue homogenized. Cytokine and chemokine levels in the lung homogenates were measured using 12-plex multiplex Luminex assay, similar trends were observed in BALF. Data are representative of two independent experiments consisting of 4–5 mice per group. Bar graphs show the group mean ± one SEM. Statistically significant differences were determined using a Kruskal-Wallis one-way ANOVA with Dunn’s post-test (*p
    Figure Legend Snippet: IL-1α signaling enhances expression of leukocyte recruiting chemokines. C57BL/6 mice treated with either isotype control antibody or IL-1α neutralizing antibody, Il1r1 -deficient and Pycard -deficient mice were infected with 5×10 7 CEA10 conidia and at 24 hours post-infection, mice were euthanized, BALF collected, and lung tissue homogenized. Cytokine and chemokine levels in the lung homogenates were measured using 12-plex multiplex Luminex assay, similar trends were observed in BALF. Data are representative of two independent experiments consisting of 4–5 mice per group. Bar graphs show the group mean ± one SEM. Statistically significant differences were determined using a Kruskal-Wallis one-way ANOVA with Dunn’s post-test (*p

    Techniques Used: Expressing, Mouse Assay, Infection, Multiplex Assay, Luminex

    C57BL/6 mice show differential expression of IL-1α and IL-1ß after A. fumigatus infection. Mice were infected i.t. with 5×10 7 CEA10 conidia and at indicated time-points, mice were euthanized, bronchoalveolar lavage fluid (BALF) collected, and lung tissue homogenized. IL-1β (A) , IL-18 (B) , IL-1α (C) , and IL-1Ra (D) levels in lung homogenate (IL-1α) and BALF (IL-1β, IL-18, and IL-1Ra) were measured using ProcartaPlex Mouse Cytokine Chemokine 36-plex Immunoassay or ELISA. Data are representative of four mice per time point and two independent experiments. Each dot represents the mean ± one SEM.
    Figure Legend Snippet: C57BL/6 mice show differential expression of IL-1α and IL-1ß after A. fumigatus infection. Mice were infected i.t. with 5×10 7 CEA10 conidia and at indicated time-points, mice were euthanized, bronchoalveolar lavage fluid (BALF) collected, and lung tissue homogenized. IL-1β (A) , IL-18 (B) , IL-1α (C) , and IL-1Ra (D) levels in lung homogenate (IL-1α) and BALF (IL-1β, IL-18, and IL-1Ra) were measured using ProcartaPlex Mouse Cytokine Chemokine 36-plex Immunoassay or ELISA. Data are representative of four mice per time point and two independent experiments. Each dot represents the mean ± one SEM.

    Techniques Used: Mouse Assay, Expressing, Infection, Enzyme-linked Immunosorbent Assay

    6) Product Images from "Enterobacter sakazakii targets DC-SIGN to induce immunosuppressive responses in dendritic cells by modulating MAP kinases"

    Article Title: Enterobacter sakazakii targets DC-SIGN to induce immunosuppressive responses in dendritic cells by modulating MAP kinases

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.0902029

    Cytokine production by DCs infected with ES
    Figure Legend Snippet: Cytokine production by DCs infected with ES

    Techniques Used: Infection

    7) Product Images from "Distinct Effects of Monophosphoryl Lipid A, Oligodeoxynucleotide CpG, and Combination Adjuvants on Modulating Innate and Adaptive Immune Responses to Influenza Vaccination"

    Article Title: Distinct Effects of Monophosphoryl Lipid A, Oligodeoxynucleotide CpG, and Combination Adjuvants on Modulating Innate and Adaptive Immune Responses to Influenza Vaccination

    Journal: Immune Network

    doi: 10.4110/in.2017.17.5.326

    Cytokines and chemokines in lung samples after lethal virus infection of mice. Lung samples were harvested from the immunized mice (n=5) day 7 post A/PR8 virus infection. Cytokine and chemokine levels of each lung samples were measured by ELISA. All results were shown in mean±SEM. For statistical analysis, One-way ANOVA and Tukey's post-multiple comparison test were performed. * p
    Figure Legend Snippet: Cytokines and chemokines in lung samples after lethal virus infection of mice. Lung samples were harvested from the immunized mice (n=5) day 7 post A/PR8 virus infection. Cytokine and chemokine levels of each lung samples were measured by ELISA. All results were shown in mean±SEM. For statistical analysis, One-way ANOVA and Tukey's post-multiple comparison test were performed. * p

    Techniques Used: Infection, Mouse Assay, Enzyme-linked Immunosorbent Assay

    Cytokine production of spleen cells from the immunized mice after in vitro antigen stimulation. Spleen cells were harvested from the immunized mice day 7 post infection and then cultured with inactivated A/PR8 virus stimulation. After 3 days culture, cytokine levels in supernatants were determined by ELISA. All results were shown in mean±SEM. For statistical analysis, One-way ANOVA and Tukey's post-multiple comparison tests were performed. nd, not detected or values below detection limit. ** p
    Figure Legend Snippet: Cytokine production of spleen cells from the immunized mice after in vitro antigen stimulation. Spleen cells were harvested from the immunized mice day 7 post infection and then cultured with inactivated A/PR8 virus stimulation. After 3 days culture, cytokine levels in supernatants were determined by ELISA. All results were shown in mean±SEM. For statistical analysis, One-way ANOVA and Tukey's post-multiple comparison tests were performed. nd, not detected or values below detection limit. ** p

    Techniques Used: Mouse Assay, In Vitro, Infection, Cell Culture, Enzyme-linked Immunosorbent Assay

    Cytokine producing T cells after immunization and lethal virus infection. The immunized mice were infected with a lethal dose (2×LD50) of A/PR8 virus after 6 weeks of immunization. Lung and BAL samples were harvested day 7 post infection. Intracellular cytokine staining was performed after incubation with MHCI and II-restricted peptides for CD8 and CD4 T cell stimulation as described in the Materials and Methods section. The cytokine producing cell numbers were calculated by multiplying cell percentages with total cell numbers. All results were shown in mean±SEM. For statistical analysis, One-way ANOVA and Tukey's post-multiple comparison tests were performed. * p
    Figure Legend Snippet: Cytokine producing T cells after immunization and lethal virus infection. The immunized mice were infected with a lethal dose (2×LD50) of A/PR8 virus after 6 weeks of immunization. Lung and BAL samples were harvested day 7 post infection. Intracellular cytokine staining was performed after incubation with MHCI and II-restricted peptides for CD8 and CD4 T cell stimulation as described in the Materials and Methods section. The cytokine producing cell numbers were calculated by multiplying cell percentages with total cell numbers. All results were shown in mean±SEM. For statistical analysis, One-way ANOVA and Tukey's post-multiple comparison tests were performed. * p

    Techniques Used: Infection, Mouse Assay, Staining, Incubation, Cell Stimulation

    In vitro proliferation and activation of T cells by adjuvant-activated DCs. DCs enriched from bone marrow cells were pre-activated by MPL (0.2 μg/ml), CpG (1 μg/ml), or MPL (0.2 μg/ml)+CpG (1 μg/ml) for 2 days. Allogeneic lymphocytes were harvested from spleens of C57BL/6 mice. CFSE-labeled lymphocytes and pre-activated DCs were co-cultured for 5 days. T cell proliferation and cytokine producing cells were determined by flow cytometry. (A) Proliferated CD4 + T cells. (B) Proliferated CD8 + T cells. (C) IFN-γ producing CD4 + T cells. (D) IFN-γ producing CD8 + T cells. (E) IL-4 producing CD4 + T cells. (F) IL-4 producing CD8 + T cells. All results were shown in mean±SEM. For statistical analysis, One-way ANOVA and Tukey's post-multiple comparison tests were performed. ns, not significant between the indicated groups. * p
    Figure Legend Snippet: In vitro proliferation and activation of T cells by adjuvant-activated DCs. DCs enriched from bone marrow cells were pre-activated by MPL (0.2 μg/ml), CpG (1 μg/ml), or MPL (0.2 μg/ml)+CpG (1 μg/ml) for 2 days. Allogeneic lymphocytes were harvested from spleens of C57BL/6 mice. CFSE-labeled lymphocytes and pre-activated DCs were co-cultured for 5 days. T cell proliferation and cytokine producing cells were determined by flow cytometry. (A) Proliferated CD4 + T cells. (B) Proliferated CD8 + T cells. (C) IFN-γ producing CD4 + T cells. (D) IFN-γ producing CD8 + T cells. (E) IL-4 producing CD4 + T cells. (F) IL-4 producing CD8 + T cells. All results were shown in mean±SEM. For statistical analysis, One-way ANOVA and Tukey's post-multiple comparison tests were performed. ns, not significant between the indicated groups. * p

    Techniques Used: In Vitro, Activation Assay, Mouse Assay, Labeling, Cell Culture, Flow Cytometry, Cytometry

    In vitro activation of bone marrow-derived DCs by adjuvant stimulation. DCs were enriched from mouse bone marrow cells by treatment with mGM-CSF. (A-C) Cytokine levels secreted into the culture supernatants of DCs treated with different concentrations of MPL and CpG were measured by ELISA. For statistical analysis, Two-way ANOVA and Bonferroni post-multiple comparison tests were performed. (D-F) The immature DCs were cultured with MPL (0.2 μg/ml), CpG (1 μg/ml), or MPL (0.2 μg/ml)+CpG (1 μg/ml) for 2 days. Expression levels of DC activation markers were determined by flow cytometry. All results were shown in mean±SEM. For statistical analysis, One-way ANOVA and Tukey's post-multiple comparison tests were performed. mGM-CSF, mouse granulocyte-macrophage colony stimulating factor. * p
    Figure Legend Snippet: In vitro activation of bone marrow-derived DCs by adjuvant stimulation. DCs were enriched from mouse bone marrow cells by treatment with mGM-CSF. (A-C) Cytokine levels secreted into the culture supernatants of DCs treated with different concentrations of MPL and CpG were measured by ELISA. For statistical analysis, Two-way ANOVA and Bonferroni post-multiple comparison tests were performed. (D-F) The immature DCs were cultured with MPL (0.2 μg/ml), CpG (1 μg/ml), or MPL (0.2 μg/ml)+CpG (1 μg/ml) for 2 days. Expression levels of DC activation markers were determined by flow cytometry. All results were shown in mean±SEM. For statistical analysis, One-way ANOVA and Tukey's post-multiple comparison tests were performed. mGM-CSF, mouse granulocyte-macrophage colony stimulating factor. * p

    Techniques Used: In Vitro, Activation Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing, Flow Cytometry, Cytometry

    8) Product Images from "Attenuation of NF-κB in intestinal epithelial cells is sufficient to mitigate the bone loss co-morbidity of experimental mouse colitis."

    Article Title: Attenuation of NF-κB in intestinal epithelial cells is sufficient to mitigate the bone loss co-morbidity of experimental mouse colitis.

    Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

    doi: 10.1002/jbmr.3759

    Expression of inflammatory cytokines in serum and IECs derived from DSS-colitis is associated with robust NF-κB activation in IECs. (A) Serum and intestinal epithelial cells were collected from control (n=6) and DSS-treated (n=6) mice. Levels of serum cytokines were measured using Millipore Luminex ELISA. (B) Small intestine was processed to extract epithelial layer-derived IECs. Q-PCR was used to measure cytokine expression in IECs. (C) Luciferase assay was performed in IECs to measure NF-κB activation. The IECs were further sorted for CD326 + cells and either lysed to measure mRNA expression of IKK2 (D) or stained to measure p-IKK2 + CD326 + IECs (E) and p-p65 + CD326 + IECs (F). Y axis in panels A and B is a log scale with breaks to accommodate presentation of low and high expressed cytokines in the same graph. *=p
    Figure Legend Snippet: Expression of inflammatory cytokines in serum and IECs derived from DSS-colitis is associated with robust NF-κB activation in IECs. (A) Serum and intestinal epithelial cells were collected from control (n=6) and DSS-treated (n=6) mice. Levels of serum cytokines were measured using Millipore Luminex ELISA. (B) Small intestine was processed to extract epithelial layer-derived IECs. Q-PCR was used to measure cytokine expression in IECs. (C) Luciferase assay was performed in IECs to measure NF-κB activation. The IECs were further sorted for CD326 + cells and either lysed to measure mRNA expression of IKK2 (D) or stained to measure p-IKK2 + CD326 + IECs (E) and p-p65 + CD326 + IECs (F). Y axis in panels A and B is a log scale with breaks to accommodate presentation of low and high expressed cytokines in the same graph. *=p

    Techniques Used: Expressing, Derivative Assay, Activation Assay, Mouse Assay, Luminex, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Luciferase, Staining

    9) Product Images from "Resolvins Decrease Oxidative Stress Mediated Macrophage and Epithelial Cell Interaction through Decreased Cytokine Secretion"

    Article Title: Resolvins Decrease Oxidative Stress Mediated Macrophage and Epithelial Cell Interaction through Decreased Cytokine Secretion

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0136755

    Resolvin Treatment leads to reduced macrophage induced alveolar epithelial cytokine secretion. THP-1 macrophages were treated with ATP + H 2 O 2 in the presence or absence of RvD1 (100 nM) and AT-RvD1 (100 nM). Following treatment, supernatant from each macrophage group was collected and used to treat A549 cells for 6 hours. After treatment, supernatant from alveolar epithelial cells was collected and analyzed for interleukin-8 via ELISA. One way ANOVA was used with a tukey post-hoc test, where a p-value
    Figure Legend Snippet: Resolvin Treatment leads to reduced macrophage induced alveolar epithelial cytokine secretion. THP-1 macrophages were treated with ATP + H 2 O 2 in the presence or absence of RvD1 (100 nM) and AT-RvD1 (100 nM). Following treatment, supernatant from each macrophage group was collected and used to treat A549 cells for 6 hours. After treatment, supernatant from alveolar epithelial cells was collected and analyzed for interleukin-8 via ELISA. One way ANOVA was used with a tukey post-hoc test, where a p-value

    Techniques Used: Enzyme-linked Immunosorbent Assay

    AT-RvD1 blunts macrophage and epithelial communication through reduced cytokine signaling. Exposure to oxidative stress leads to an enhanced secretion of proinflammatory cytokines, with IL-1β being the most bioactive for ALI patients. IL-1β secretion results in alveolar epithelial cell activation which is hallmarked by enhanced barrier function, cytokine secretion, and adhesion molecule expression. We found that AT-RvD1 was able to interrupt the macrophage to alveolar communication through the blockage of IL-1β signaling. Upon treatment of alveolar epithelial cells with IL-1β, there was an increase in inflammatory activation, which was significantly attenuated with AT-RvD1 treatment.
    Figure Legend Snippet: AT-RvD1 blunts macrophage and epithelial communication through reduced cytokine signaling. Exposure to oxidative stress leads to an enhanced secretion of proinflammatory cytokines, with IL-1β being the most bioactive for ALI patients. IL-1β secretion results in alveolar epithelial cell activation which is hallmarked by enhanced barrier function, cytokine secretion, and adhesion molecule expression. We found that AT-RvD1 was able to interrupt the macrophage to alveolar communication through the blockage of IL-1β signaling. Upon treatment of alveolar epithelial cells with IL-1β, there was an increase in inflammatory activation, which was significantly attenuated with AT-RvD1 treatment.

    Techniques Used: Activation Assay, Expressing

    Aspirin-Triggered Resolvin D1 Attenuated IL-1β-induced Cytokine/Chemokine Secretion. A549 cells were seeded at a density of 0.5 x 10 6 million cells per well in 12 well plates. When cells reached confluence, they were then serum starved and treated with IL-1β (10 ng/mL) in the presence or absence of aspirin-triggered resolvin D1 (AT-RD1, 100 nM) for 6 hours. Following treatment, cell culture supernatants were collected and the presence of (A) IL-8 and (B) IL-6 levels were analyzed by ELISA. A student t-test was used to determine statistical significance with p
    Figure Legend Snippet: Aspirin-Triggered Resolvin D1 Attenuated IL-1β-induced Cytokine/Chemokine Secretion. A549 cells were seeded at a density of 0.5 x 10 6 million cells per well in 12 well plates. When cells reached confluence, they were then serum starved and treated with IL-1β (10 ng/mL) in the presence or absence of aspirin-triggered resolvin D1 (AT-RD1, 100 nM) for 6 hours. Following treatment, cell culture supernatants were collected and the presence of (A) IL-8 and (B) IL-6 levels were analyzed by ELISA. A student t-test was used to determine statistical significance with p

    Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay

    10) Product Images from "A water extract of Samchulkunbi-tang attenuates airway inflammation by inhibiting inos and MMP-9 activities in an ovalbumin-induced murine asthma model"

    Article Title: A water extract of Samchulkunbi-tang attenuates airway inflammation by inhibiting inos and MMP-9 activities in an ovalbumin-induced murine asthma model

    Journal: BMC Complementary and Alternative Medicine

    doi: 10.1186/1472-6882-12-257

    Effects of SCTE on cytokine and chemokine levels in BALF. BALF was collected from mice 48 h after the last OVA challenge. Individual samples were analyzed using ELISA. ( A ) IL-4; ( B ) IL-13; ( C ) IL-33, ( D ) TNF-α, ( E ) eotaxin. PBS/PBS, PBS-sensitized/challenged, negative control (PBS only); OVA/PBS, OVA-sensitized/challenged mice (PBS only); OVA/mon, OVA-sensitized/challenged mice (montelukast 30 mg/kg); OVA/SCTE-200, OVA-sensitized/challenged mice (SCTE 200 mg/kg); OVA/SCTE-400, OVA-sensitized/challenged mice (SCTE 400 mg/kg). SCTE or montelukast was given 1 h before the challenge. Significantly different from PBS/PBS, ## P
    Figure Legend Snippet: Effects of SCTE on cytokine and chemokine levels in BALF. BALF was collected from mice 48 h after the last OVA challenge. Individual samples were analyzed using ELISA. ( A ) IL-4; ( B ) IL-13; ( C ) IL-33, ( D ) TNF-α, ( E ) eotaxin. PBS/PBS, PBS-sensitized/challenged, negative control (PBS only); OVA/PBS, OVA-sensitized/challenged mice (PBS only); OVA/mon, OVA-sensitized/challenged mice (montelukast 30 mg/kg); OVA/SCTE-200, OVA-sensitized/challenged mice (SCTE 200 mg/kg); OVA/SCTE-400, OVA-sensitized/challenged mice (SCTE 400 mg/kg). SCTE or montelukast was given 1 h before the challenge. Significantly different from PBS/PBS, ## P

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Negative Control

    11) Product Images from "IQ Domain-Containing GTPase-Activating Protein 1 Regulates Cytoskeletal Reorganization and Facilitates NKG2D-Mediated Mechanistic Target of Rapamycin Complex 1 Activation and Cytokine Gene Translation in Natural Killer Cells"

    Article Title: IQ Domain-Containing GTPase-Activating Protein 1 Regulates Cytoskeletal Reorganization and Facilitates NKG2D-Mediated Mechanistic Target of Rapamycin Complex 1 Activation and Cytokine Gene Translation in Natural Killer Cells

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.01168

    IQ domain-containing GTPase-activating protein 1 (IQGAP1) regulates inflammatory cytokine production in natural killer (NK) cells. (A) Cytokine (IFN-γ and GM-CSF) and chemokine (CCL3, CCL4, and CCL5) production by wild-type (WT) and Iqgap1 −/− NK cells were assessed 18–21 h post-NKR stimulation (2 µg/ml) by BioPlex mouse cytokine assay. (B,C) Intracellular IFN-γ and (D,E) CCL3 accumulation was evaluated by flow cytometry 12-h post-NKG2D stimulation (2 µg/ml), and the data are quantified as percent positive NK cells (CD3ε − NK1.1 + ). (F) Cytokine and chemokine production by WT and Iqgap1 −/− NK cells was assessed 18–21 after the addition of IL-12 (1 ng/ml) and IL-18 (10 ng/ml) by BioPlex mouse cytokine assay. Error bars represent SEM (A,F) or SD (C,E) using five to eight mice in at least two independent experiments, * p
    Figure Legend Snippet: IQ domain-containing GTPase-activating protein 1 (IQGAP1) regulates inflammatory cytokine production in natural killer (NK) cells. (A) Cytokine (IFN-γ and GM-CSF) and chemokine (CCL3, CCL4, and CCL5) production by wild-type (WT) and Iqgap1 −/− NK cells were assessed 18–21 h post-NKR stimulation (2 µg/ml) by BioPlex mouse cytokine assay. (B,C) Intracellular IFN-γ and (D,E) CCL3 accumulation was evaluated by flow cytometry 12-h post-NKG2D stimulation (2 µg/ml), and the data are quantified as percent positive NK cells (CD3ε − NK1.1 + ). (F) Cytokine and chemokine production by WT and Iqgap1 −/− NK cells was assessed 18–21 after the addition of IL-12 (1 ng/ml) and IL-18 (10 ng/ml) by BioPlex mouse cytokine assay. Error bars represent SEM (A,F) or SD (C,E) using five to eight mice in at least two independent experiments, * p

    Techniques Used: Cytokine Assay, Flow Cytometry, Cytometry, Mouse Assay

    NKG2D signaling and induction of cytokine gene transcripts in Iqgap1 −/− natural killer (NK) cells. (A) Phosphorylation of Akt at Ser 473 and Thr 308 was determined by western blot in NKG2D-stimulated wild-type (WT) and Iqgap1 −/− NK cells at 20- and 60-min post-NKG2D activation and total Akt was used as the loading control (B) Degradation of IκBα is shown after activation with NKG2D in WT and Iqgap1 −/− NK cells at indicated times post-activation with β-actin as the loading control. (C) Phosphorylation of mitogen-activated protein kinases, Erk1/2 (Thr 202 and Tyr 204 ), and Jnk1/2 (Thr 183 and Tyr 185 ), in WT and Iqgap1 −/− NK cells are shown at indicated times post-NKG2D activation with total Erk1/2 and Jnk1/2 proteins serving loading controls. (D) Fold induction of Ifng transcript, relative to unstimulated WT NK cells, was determined by RT-qPCR 4-h post-NKG2D stimulation. (E) Microarray data represented by a scatter plot using a 3-Log2 fold change cutoff. NKG2D-induced cytokine transcripts are labeled. (F) Changes in the induction of solute carrier transcripts. Those that directly regulate amino acid transport are highlighted green. Error bars represent SD using four to five mice in at least two independent experiments (A–D) or four mice of each genotype pooled in one experiment (E,F) .
    Figure Legend Snippet: NKG2D signaling and induction of cytokine gene transcripts in Iqgap1 −/− natural killer (NK) cells. (A) Phosphorylation of Akt at Ser 473 and Thr 308 was determined by western blot in NKG2D-stimulated wild-type (WT) and Iqgap1 −/− NK cells at 20- and 60-min post-NKG2D activation and total Akt was used as the loading control (B) Degradation of IκBα is shown after activation with NKG2D in WT and Iqgap1 −/− NK cells at indicated times post-activation with β-actin as the loading control. (C) Phosphorylation of mitogen-activated protein kinases, Erk1/2 (Thr 202 and Tyr 204 ), and Jnk1/2 (Thr 183 and Tyr 185 ), in WT and Iqgap1 −/− NK cells are shown at indicated times post-NKG2D activation with total Erk1/2 and Jnk1/2 proteins serving loading controls. (D) Fold induction of Ifng transcript, relative to unstimulated WT NK cells, was determined by RT-qPCR 4-h post-NKG2D stimulation. (E) Microarray data represented by a scatter plot using a 3-Log2 fold change cutoff. NKG2D-induced cytokine transcripts are labeled. (F) Changes in the induction of solute carrier transcripts. Those that directly regulate amino acid transport are highlighted green. Error bars represent SD using four to five mice in at least two independent experiments (A–D) or four mice of each genotype pooled in one experiment (E,F) .

    Techniques Used: Western Blot, Activation Assay, Quantitative RT-PCR, Microarray, Labeling, Mouse Assay

    12) Product Images from "Three Weeks of Murine Hindlimb Unloading Induces Shifts from B to T and from Th to Tc Splenic Lymphocytes in Absence of Stress and Differentially Reduces Cell-Specific Mitogenic Responses"

    Article Title: Three Weeks of Murine Hindlimb Unloading Induces Shifts from B to T and from Th to Tc Splenic Lymphocytes in Absence of Stress and Differentially Reduces Cell-Specific Mitogenic Responses

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0092664

    Cytokines secreted by splenic lymphocytes stimulated with LPS. Splenocytes were incubated with 5 μg/ml of LPS for 72 h. Cytokine concentrations in the supernatants were determined by flow cytometry using a Flowcytomix kit. Each group, hindlimb unloaded (HU), restrained (R) and control (C), was compared to the others. n = 5 mice per group. No significant difference was found between the three experimental groups using ANOVA and Tukey post-hoc test. Cytokines whose concentrations were below the detection threshold of the kit are not indicated.
    Figure Legend Snippet: Cytokines secreted by splenic lymphocytes stimulated with LPS. Splenocytes were incubated with 5 μg/ml of LPS for 72 h. Cytokine concentrations in the supernatants were determined by flow cytometry using a Flowcytomix kit. Each group, hindlimb unloaded (HU), restrained (R) and control (C), was compared to the others. n = 5 mice per group. No significant difference was found between the three experimental groups using ANOVA and Tukey post-hoc test. Cytokines whose concentrations were below the detection threshold of the kit are not indicated.

    Techniques Used: Incubation, Flow Cytometry, Cytometry, Mouse Assay

    Cytokines secreted by splenic lymphocytes stimulated with ConA. Splenocytes were incubated with 5 μg/ml of ConA for 72 h. Cytokine concentrations in the supernatants were determined by flow cytometry using a Flowcytomix kit. Each group, hindlimb unloaded (HU), restrained (R) and control (C), was compared to the others. n = 5 mice per group. No significant difference was found between the three experimental groups using ANOVA and Tukey post-hoc test. Cytokines whose concentrations were below the detection threshold of the kit are not indicated.
    Figure Legend Snippet: Cytokines secreted by splenic lymphocytes stimulated with ConA. Splenocytes were incubated with 5 μg/ml of ConA for 72 h. Cytokine concentrations in the supernatants were determined by flow cytometry using a Flowcytomix kit. Each group, hindlimb unloaded (HU), restrained (R) and control (C), was compared to the others. n = 5 mice per group. No significant difference was found between the three experimental groups using ANOVA and Tukey post-hoc test. Cytokines whose concentrations were below the detection threshold of the kit are not indicated.

    Techniques Used: Incubation, Flow Cytometry, Cytometry, Mouse Assay

    13) Product Images from "The Immunomodulatory Activity of Meningococcal Lipoprotein Ag473 Depends on the Conformation Made up of the Lipid and Protein Moieties"

    Article Title: The Immunomodulatory Activity of Meningococcal Lipoprotein Ag473 Depends on the Conformation Made up of the Lipid and Protein Moieties

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0040873

    L-Ag473 induces cytokine and chemokine production by BMDCs. BMDCs were incubated with L-Ag473 for 24 hours (or 6 hours for TNF-α and RANTES). Supernatants were collected and (A) TNF-α, IL-6, and IL-12; (B) MCP-1, MIP-1, and RANTES were determined by ELISA. Data are shown as mean ± SD from triplicate DC cultures; NS p > 0.05; * p
    Figure Legend Snippet: L-Ag473 induces cytokine and chemokine production by BMDCs. BMDCs were incubated with L-Ag473 for 24 hours (or 6 hours for TNF-α and RANTES). Supernatants were collected and (A) TNF-α, IL-6, and IL-12; (B) MCP-1, MIP-1, and RANTES were determined by ELISA. Data are shown as mean ± SD from triplicate DC cultures; NS p > 0.05; * p

    Techniques Used: Incubation, Enzyme-linked Immunosorbent Assay

    14) Product Images from "Zika Virus infection of rhesus macaques leads to viral persistence in multiple tissues"

    Article Title: Zika Virus infection of rhesus macaques leads to viral persistence in multiple tissues

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006219

    Rhesus cytokine and chemokine production in response to ZIKV infection and block of NF-kB signaling in Rhesus fibroblasts. A 29-plex-cytokine/chemokine/growth factor magnetic bead assay was performed on plasma from rhesus monkeys at all time points post infection. Cytokine analysis revealed changes in only A) IL-RA; B) MCP-1-CCL2; C) IP-10-CXCL10; and D) ITAC-CXCL11. Reporter assay showing induction of NF-κB-dependent (E, F) or interferon stimulated response element (ISRE)-dependent (G) LUC expression in fibroblasts infected for 56h with ZIKV at MOI = 5ffu/cell. Luminescence was measured 8h after treatment with 60μg/mL poly(I:C) (E) 100ng/mL human IL-1β (F) or 5,000 units/ml IFNβ1 (G). Values displayed are average fold changes (three replicates) of stimulated versus untreated cells ±SD.
    Figure Legend Snippet: Rhesus cytokine and chemokine production in response to ZIKV infection and block of NF-kB signaling in Rhesus fibroblasts. A 29-plex-cytokine/chemokine/growth factor magnetic bead assay was performed on plasma from rhesus monkeys at all time points post infection. Cytokine analysis revealed changes in only A) IL-RA; B) MCP-1-CCL2; C) IP-10-CXCL10; and D) ITAC-CXCL11. Reporter assay showing induction of NF-κB-dependent (E, F) or interferon stimulated response element (ISRE)-dependent (G) LUC expression in fibroblasts infected for 56h with ZIKV at MOI = 5ffu/cell. Luminescence was measured 8h after treatment with 60μg/mL poly(I:C) (E) 100ng/mL human IL-1β (F) or 5,000 units/ml IFNβ1 (G). Values displayed are average fold changes (three replicates) of stimulated versus untreated cells ±SD.

    Techniques Used: Infection, Blocking Assay, Reporter Assay, Expressing

    15) Product Images from "CD4+CD25+ Regulatory Cells Contribute to the Regulation of Colonic Th2 Granulomatous Pathology Caused by Schistosome Infection"

    Article Title: CD4+CD25+ Regulatory Cells Contribute to the Regulation of Colonic Th2 Granulomatous Pathology Caused by Schistosome Infection

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0001269

    Egg antigen-specific Th2, but not TGF-β1 responses become down-modulated within the mLN and colon during chronic infection. A) Proliferative responses and B) cytokine release (pg/ml) by mLN cells from naïve, acute, or chronic mice (n = 4/group) to anti-CD3 mAb, or SEA. C) Cytokine levels within colonic tissues (pg/mg tissue) and D) adjusted for numbers of eggs / mg of tissue. Data are mean proliferative response / cytokine concentration ±SEM.
    Figure Legend Snippet: Egg antigen-specific Th2, but not TGF-β1 responses become down-modulated within the mLN and colon during chronic infection. A) Proliferative responses and B) cytokine release (pg/ml) by mLN cells from naïve, acute, or chronic mice (n = 4/group) to anti-CD3 mAb, or SEA. C) Cytokine levels within colonic tissues (pg/mg tissue) and D) adjusted for numbers of eggs / mg of tissue. Data are mean proliferative response / cytokine concentration ±SEM.

    Techniques Used: Infection, Mouse Assay, Concentration Assay

    Transfer of schistosome-expanded CD4 + CD25 + T regs modulates the development of acute-stage granulomas. A) Isolated mLN CD4 + CD25 + T regs from mice with chronic infection used for transfer. B) 3D images of multiphoton confocal stacks of colonic tissue viewed in situ at the acute stage of infection in hCD2-VaDsRed-B.6 control mice, or recipients of infection-expanded CD4 + CD25 + T regs (2.5✕10 6 ). Grid squares are 63.9 µm2. C) Granuloma area (left), DsRed + cell counts (middle), and collagen half-volume (right) taken from confocal stacks (as above). Data are from 5–6 separate granulomas per mouse. Bars are mean / group (n = 3). D) Soluble collagen and cytokine in colonic extracts from infected control mice, or recipients of CD4 + CD25 + cells. Bars are means / group (n = 3). Data is mean pg/ml (± SEM) per group.
    Figure Legend Snippet: Transfer of schistosome-expanded CD4 + CD25 + T regs modulates the development of acute-stage granulomas. A) Isolated mLN CD4 + CD25 + T regs from mice with chronic infection used for transfer. B) 3D images of multiphoton confocal stacks of colonic tissue viewed in situ at the acute stage of infection in hCD2-VaDsRed-B.6 control mice, or recipients of infection-expanded CD4 + CD25 + T regs (2.5✕10 6 ). Grid squares are 63.9 µm2. C) Granuloma area (left), DsRed + cell counts (middle), and collagen half-volume (right) taken from confocal stacks (as above). Data are from 5–6 separate granulomas per mouse. Bars are mean / group (n = 3). D) Soluble collagen and cytokine in colonic extracts from infected control mice, or recipients of CD4 + CD25 + cells. Bars are means / group (n = 3). Data is mean pg/ml (± SEM) per group.

    Techniques Used: Isolation, Mouse Assay, Infection, In Situ

    16) Product Images from "Role of human Pegivirus infections in whole Plasmodium falciparum sporozoite vaccination and controlled human malaria infection in African volunteers"

    Article Title: Role of human Pegivirus infections in whole Plasmodium falciparum sporozoite vaccination and controlled human malaria infection in African volunteers

    Journal: Virology Journal

    doi: 10.1186/s12985-021-01500-8

    HPgV-1 infection is associated with increased systemic levels of IL-2 and IL-17A. Cytokine, chemokine and growth factors levels were analysed by Luminex and levels compared between HPgV-1 negative (5′ UTR- , grey, n = 35 ) and HPgV-1 positive (5′ UTR + , purple, n = 9) volunteers. Absolute serum concentrations levels (pg/ml) of Interluekin-2 (IL-2) and Interluekin-17A (IL-17A) at samples taken before vaccination are shown. Significantly higher IL-2 and IL-17A are seen in the HPgV-1 + compared to the HPgV-1 − . Wilcoxon rank sum test was used to determine significance ( P value *
    Figure Legend Snippet: HPgV-1 infection is associated with increased systemic levels of IL-2 and IL-17A. Cytokine, chemokine and growth factors levels were analysed by Luminex and levels compared between HPgV-1 negative (5′ UTR- , grey, n = 35 ) and HPgV-1 positive (5′ UTR + , purple, n = 9) volunteers. Absolute serum concentrations levels (pg/ml) of Interluekin-2 (IL-2) and Interluekin-17A (IL-17A) at samples taken before vaccination are shown. Significantly higher IL-2 and IL-17A are seen in the HPgV-1 + compared to the HPgV-1 − . Wilcoxon rank sum test was used to determine significance ( P value *

    Techniques Used: Infection, Luminex

    17) Product Images from "Biological Characterization of Lipopolysaccharide from Treponema pectinovorum"

    Article Title: Biological Characterization of Lipopolysaccharide from Treponema pectinovorum

    Journal: Infection and Immunity

    doi: 10.1128/IAI.70.1.211-217.2002

    Cytokine (IL-1β, TNF-α, and IL-6) pattern in C3H/HeN Tac-MTV (LPS responder) mice after LPS treatment. Mice ( n = 3) were injected i.p. with 5, 0.5, and 0.05 μg of T. pectinovorum ATCC 33768 LPS with GalN as indicated in Materials and Methods. Serum samples were taken at 3, 8, and 12 h after LPS administration. Each cytokine concentration was determined by ELISA. Results are the means and standard deviations for three mice at 12 h.
    Figure Legend Snippet: Cytokine (IL-1β, TNF-α, and IL-6) pattern in C3H/HeN Tac-MTV (LPS responder) mice after LPS treatment. Mice ( n = 3) were injected i.p. with 5, 0.5, and 0.05 μg of T. pectinovorum ATCC 33768 LPS with GalN as indicated in Materials and Methods. Serum samples were taken at 3, 8, and 12 h after LPS administration. Each cytokine concentration was determined by ELISA. Results are the means and standard deviations for three mice at 12 h.

    Techniques Used: Mouse Assay, Injection, Concentration Assay, Enzyme-linked Immunosorbent Assay

    Proinflammatory-cytokine (IL-1β, TNF-α, and IL-6) patterns in C3H/HeN Tac-MTV (LPS responder) mice were analyzed after i.p. challenge infection with live (10 8 cells) and heat-killed (H-K) (10 8 cells) T. pectinovorum ATCC 33768 and live E. coli O111 (10 5 cells) with and without GalN as described in Materials and Methods. Serum samples were taken at 3, 8, and 12 h postinfection. Each cytokine concentration was determined by sequential ELISA. Results are the means and standard deviations for three mice at 12 h.
    Figure Legend Snippet: Proinflammatory-cytokine (IL-1β, TNF-α, and IL-6) patterns in C3H/HeN Tac-MTV (LPS responder) mice were analyzed after i.p. challenge infection with live (10 8 cells) and heat-killed (H-K) (10 8 cells) T. pectinovorum ATCC 33768 and live E. coli O111 (10 5 cells) with and without GalN as described in Materials and Methods. Serum samples were taken at 3, 8, and 12 h postinfection. Each cytokine concentration was determined by sequential ELISA. Results are the means and standard deviations for three mice at 12 h.

    Techniques Used: Mouse Assay, Infection, Concentration Assay, Enzyme-linked Immunosorbent Assay

    18) Product Images from "Induction of Inflammatory Responses by Carbon Fullerene (C60) in Cultured RAW264.7 Cells and in Intraperitoneally Injected Mice"

    Article Title: Induction of Inflammatory Responses by Carbon Fullerene (C60) in Cultured RAW264.7 Cells and in Intraperitoneally Injected Mice

    Journal: Toxicological Research

    doi: 10.5487/TR.2010.26.4.267

    Effects of C60s on the expression of cytokine genes by a single peritoneal injection. Mice were intraperitoneally injected with fullerene 2mg/kg bw and were sacrificed at 6 24 48 and 72 hrs after injection (n = 6) . The peritoneal fluid was harvested and pooled by 2 mice to make 3 test samples (total 6 mice per group at each time point) at the designated time after injection.After isolation of peritoneal macrophage RNA was extracted from the cells and amplified by RT-PCR using the respective primers described in Table 1 . The results were confirmed by several separate experiments and representative images are shown.
    Figure Legend Snippet: Effects of C60s on the expression of cytokine genes by a single peritoneal injection. Mice were intraperitoneally injected with fullerene 2mg/kg bw and were sacrificed at 6 24 48 and 72 hrs after injection (n = 6) . The peritoneal fluid was harvested and pooled by 2 mice to make 3 test samples (total 6 mice per group at each time point) at the designated time after injection.After isolation of peritoneal macrophage RNA was extracted from the cells and amplified by RT-PCR using the respective primers described in Table 1 . The results were confirmed by several separate experiments and representative images are shown.

    Techniques Used: Expressing, Injection, Mouse Assay, Isolation, Amplification, Reverse Transcription Polymerase Chain Reaction

    19) Product Images from "Comparison of ophthalmic sponges and extraction buffers for quantifying cytokine profiles in tears using Luminex technology"

    Article Title: Comparison of ophthalmic sponges and extraction buffers for quantifying cytokine profiles in tears using Luminex technology

    Journal: Molecular Vision

    doi:

    Mean percentages of cytokine/chemokine recovery from the Merocel sponge (gray columns) loaded in vitro with known concentration of 25 cytokines/chemokines and extracted with an extraction buffer 1 (EX1).
    Figure Legend Snippet: Mean percentages of cytokine/chemokine recovery from the Merocel sponge (gray columns) loaded in vitro with known concentration of 25 cytokines/chemokines and extracted with an extraction buffer 1 (EX1).

    Techniques Used: In Vitro, Concentration Assay

    Mean percentages of cytokine/chemokine recovery from Merocel (black columns), Pro-ophta (white columns) and Weck-Cel (gray columns) sponges loaded in vitro with known concentration of 25 cytokines/chemokines and extracted with assay diluent used as an extraction buffer (EX4).
    Figure Legend Snippet: Mean percentages of cytokine/chemokine recovery from Merocel (black columns), Pro-ophta (white columns) and Weck-Cel (gray columns) sponges loaded in vitro with known concentration of 25 cytokines/chemokines and extracted with assay diluent used as an extraction buffer (EX4).

    Techniques Used: In Vitro, Concentration Assay

    The percentages of cytokine/chemokine recovery from the Merocel (down triangle), Pro-ophta (square) and Weck-Cel sponges (star). Values for the extraction buffers (EX1-EX7) were pooled to visualize the impact of sponge type on the cytokine recovery.
    Figure Legend Snippet: The percentages of cytokine/chemokine recovery from the Merocel (down triangle), Pro-ophta (square) and Weck-Cel sponges (star). Values for the extraction buffers (EX1-EX7) were pooled to visualize the impact of sponge type on the cytokine recovery.

    Techniques Used:

    20) Product Images from "Effects of anticancer agents on cell viability, proliferative activity and cytokine production of peripheral blood mononuclear cells"

    Article Title: Effects of anticancer agents on cell viability, proliferative activity and cytokine production of peripheral blood mononuclear cells

    Journal: Journal of Clinical Biochemistry and Nutrition

    doi: 10.3164/jcbn.12-60

    Effects of GEM on PHA-induced cytokine production. After 2 h chemical stimulation with GEM, PBMCs were then incubated with PHA for 48 h. Cell culture supernatant was assayed for TNF-α(a), IL-2(b), IL-4(c), IL-10(d) and IFN-γ(e) by using a cytokine-specific solid phase sandwich ELISA. In all panels, data are expressed as the mean ± SEM. * p
    Figure Legend Snippet: Effects of GEM on PHA-induced cytokine production. After 2 h chemical stimulation with GEM, PBMCs were then incubated with PHA for 48 h. Cell culture supernatant was assayed for TNF-α(a), IL-2(b), IL-4(c), IL-10(d) and IFN-γ(e) by using a cytokine-specific solid phase sandwich ELISA. In all panels, data are expressed as the mean ± SEM. * p

    Techniques Used: Incubation, Cell Culture, Sandwich ELISA

    Effects of 5-FU on PHA-induced cytokine production. After 2 h chemical stimulation with 5-FU, PBMCs were then incubated with PHA for 48 h. Cell culture supernatant was assayed for TNF-α(a), IL-2(b), IL-4(c), IL-10(d) and IFN-γ(e) by using a cytokine-specific solid phase sandwich ELISA. In all panels, data are expressed as the mean ± SEM. * p
    Figure Legend Snippet: Effects of 5-FU on PHA-induced cytokine production. After 2 h chemical stimulation with 5-FU, PBMCs were then incubated with PHA for 48 h. Cell culture supernatant was assayed for TNF-α(a), IL-2(b), IL-4(c), IL-10(d) and IFN-γ(e) by using a cytokine-specific solid phase sandwich ELISA. In all panels, data are expressed as the mean ± SEM. * p

    Techniques Used: Incubation, Cell Culture, Sandwich ELISA

    Effects of CDDP on PHA-induced cytokine production. After 2 h chemical stimulation with CDDP, PBMCs were then incubated with PHA for 48 h. Cell culture supernatant was assayed for TNF-α(a), IL-2(b), IL-4(c), IL-10(d) and IFN-γ(e) by using a cytokine-specific solid phase sandwich ELISA. In all panels, data are expressed as the mean ± SEM. * p
    Figure Legend Snippet: Effects of CDDP on PHA-induced cytokine production. After 2 h chemical stimulation with CDDP, PBMCs were then incubated with PHA for 48 h. Cell culture supernatant was assayed for TNF-α(a), IL-2(b), IL-4(c), IL-10(d) and IFN-γ(e) by using a cytokine-specific solid phase sandwich ELISA. In all panels, data are expressed as the mean ± SEM. * p

    Techniques Used: Incubation, Cell Culture, Sandwich ELISA

    Effects of CPT-11 on PHA-induced cytokine production. After 2 h chemical stimulation with CPT-11, PBMCs were then incubated with PHA for 48 h. Cell culture supernatant was assayed for TNF-α(a), IL-2(b), IL-4(c), IL-10(d) and IFN-γ(e) by using a cytokine-specific solid phase sandwich ELISA. In all panels, data are expressed as the mean ± SEM. * p
    Figure Legend Snippet: Effects of CPT-11 on PHA-induced cytokine production. After 2 h chemical stimulation with CPT-11, PBMCs were then incubated with PHA for 48 h. Cell culture supernatant was assayed for TNF-α(a), IL-2(b), IL-4(c), IL-10(d) and IFN-γ(e) by using a cytokine-specific solid phase sandwich ELISA. In all panels, data are expressed as the mean ± SEM. * p

    Techniques Used: Cycling Probe Technology, Incubation, Cell Culture, Sandwich ELISA

    21) Product Images from "Treatment with cyclophosphamide supported by various dendritic cell-based vaccines induces diversification in CD4+ T cell response against MC38 colon carcinoma"

    Article Title: Treatment with cyclophosphamide supported by various dendritic cell-based vaccines induces diversification in CD4+ T cell response against MC38 colon carcinoma

    Journal: International Journal of Oncology

    doi: 10.3892/ijo.2015.3278

    Cytokine production by CD4 + cells isolated from tumor tissue. Tumor tissues were collected from mice (N=3–6 per group, two independent experiments). The CD4 + cells were isolated from tumor tissue using anti-CD4 antibody-coated magnetic beads. Cells were cultured for 48 h with Con A. The concentrations of IFN-γ, IL-4 and IL-10 in cell culture supernatants were measured by ELISA. Differences by mean ± SD were estimated. The statistical significance was calculated (for details see Fig. 3 ).
    Figure Legend Snippet: Cytokine production by CD4 + cells isolated from tumor tissue. Tumor tissues were collected from mice (N=3–6 per group, two independent experiments). The CD4 + cells were isolated from tumor tissue using anti-CD4 antibody-coated magnetic beads. Cells were cultured for 48 h with Con A. The concentrations of IFN-γ, IL-4 and IL-10 in cell culture supernatants were measured by ELISA. Differences by mean ± SD were estimated. The statistical significance was calculated (for details see Fig. 3 ).

    Techniques Used: Isolation, Mouse Assay, Magnetic Beads, Cell Culture, Enzyme-linked Immunosorbent Assay

    Cytokine production by spleen cells. On the 31st, 38th and 45th day of the experiments, spleens were obtained from CY ± DC-based vaccine-treated mice (N=3–6 per group, two independent experiments). Splenocytes were cultured in a presence of Con A (0.5 μg/ml). After 48 h, cell culture supernatants were collected, and the concentrations of IFN-γ, IL-4 and IL-10 were measured by ELISA (mean ± SD). The statistical significance was calculated (for details see Fig. 3 ).
    Figure Legend Snippet: Cytokine production by spleen cells. On the 31st, 38th and 45th day of the experiments, spleens were obtained from CY ± DC-based vaccine-treated mice (N=3–6 per group, two independent experiments). Splenocytes were cultured in a presence of Con A (0.5 μg/ml). After 48 h, cell culture supernatants were collected, and the concentrations of IFN-γ, IL-4 and IL-10 were measured by ELISA (mean ± SD). The statistical significance was calculated (for details see Fig. 3 ).

    Techniques Used: Mouse Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

    22) Product Images from "An unfolded variant of the major peanut allergen Ara h 2 with decreased anaphylactic potential"

    Article Title: An unfolded variant of the major peanut allergen Ara h 2 with decreased anaphylactic potential

    Journal: Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology

    doi: 10.1111/cea.12031

    Alkylation influences the response of allergen-primed mouse splenocytes ex vivo . Spleen cells of mice sensitized with untreated Ara h 2 and intravenously challenged with either untreated (untr.; left column; n = 7) or reduced/alkylated (r/a; right column, n = 7) Ara h 2 were isolated and stimulated ex vivo with plain culture medium, untreated or r/a Ara h 2, r/a ovomucoid (Ovo) or concanavalin A (Con A). (a) Activity assay: The metabolic activity of allergen-stimulated splenocytes was assessed using an XTT assay and compared to splenocytes stimulated with plain culture medium (= 100%). (b–d) Cytokine release: Supernatant of spleen cells was collected after 72 h of cultivation. Released (b) IL-4, (c) IL-13 and (d) IFNγ were analysed by ELISA. Mann–Whitney U -test; ***: P -value ≤ 0.001; **: P -value 0.001–0.01; *: P -value 0.01–0.05; ns = not significant.
    Figure Legend Snippet: Alkylation influences the response of allergen-primed mouse splenocytes ex vivo . Spleen cells of mice sensitized with untreated Ara h 2 and intravenously challenged with either untreated (untr.; left column; n = 7) or reduced/alkylated (r/a; right column, n = 7) Ara h 2 were isolated and stimulated ex vivo with plain culture medium, untreated or r/a Ara h 2, r/a ovomucoid (Ovo) or concanavalin A (Con A). (a) Activity assay: The metabolic activity of allergen-stimulated splenocytes was assessed using an XTT assay and compared to splenocytes stimulated with plain culture medium (= 100%). (b–d) Cytokine release: Supernatant of spleen cells was collected after 72 h of cultivation. Released (b) IL-4, (c) IL-13 and (d) IFNγ were analysed by ELISA. Mann–Whitney U -test; ***: P -value ≤ 0.001; **: P -value 0.001–0.01; *: P -value 0.01–0.05; ns = not significant.

    Techniques Used: Ex Vivo, Mouse Assay, Acetylene Reduction Assay, Isolation, Activity Assay, XTT Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    . (b) Human T cell line proliferation: Ara h 2-specific human T cell lines (TCLs) were stimulated with untreated (untr.) or reduced/alkylated (r/a) Ara h 2, or the respective core epitope peptide. Proliferation was measured by 3 H-labelled thymidine uptake and data were expressed as stimulation indices (SI: fold increase in thymidine uptake by stimulated cells over cells cultured in plain medium; please see also the 3 ). The letters under the TCL name indicate the core peptide specificity as shown in (a). t -test (two-tailed) statistical analysis between samples stimulated by untreated and reduced/alkylated Ara h 2; samples without asterisk were not significantly different; (c–e) Human TCL cytokine mRNA expression: TCLs were stimulated as described in (b) and mRNA was isolated and processed. Expression levels of genes coding for (c) IL-5, (d) IL-13 and (e) IFNγ were analysed by real-time PCR. The expression fold change in mRNA levels for stimulated as compared to unstimulated cells was calculated.
    Figure Legend Snippet: . (b) Human T cell line proliferation: Ara h 2-specific human T cell lines (TCLs) were stimulated with untreated (untr.) or reduced/alkylated (r/a) Ara h 2, or the respective core epitope peptide. Proliferation was measured by 3 H-labelled thymidine uptake and data were expressed as stimulation indices (SI: fold increase in thymidine uptake by stimulated cells over cells cultured in plain medium; please see also the 3 ). The letters under the TCL name indicate the core peptide specificity as shown in (a). t -test (two-tailed) statistical analysis between samples stimulated by untreated and reduced/alkylated Ara h 2; samples without asterisk were not significantly different; (c–e) Human TCL cytokine mRNA expression: TCLs were stimulated as described in (b) and mRNA was isolated and processed. Expression levels of genes coding for (c) IL-5, (d) IL-13 and (e) IFNγ were analysed by real-time PCR. The expression fold change in mRNA levels for stimulated as compared to unstimulated cells was calculated.

    Techniques Used: Acetylene Reduction Assay, Cell Culture, Two Tailed Test, Expressing, Isolation, Real-time Polymerase Chain Reaction

    23) Product Images from "Strategy for the Generation of Engineered Bone Constructs Based on Umbilical Cord Mesenchymal Stromal Cells Expanded with Human Platelet Lysate"

    Article Title: Strategy for the Generation of Engineered Bone Constructs Based on Umbilical Cord Mesenchymal Stromal Cells Expanded with Human Platelet Lysate

    Journal: Stem Cells International

    doi: 10.1155/2019/7198215

    Impact of the use of human platelet lysate (hPL) as a medium supplement for cell culture of UC-MSC. (a) Proliferation kinetics of UC-MSC as measured by population doubling time (hours), population doubling levels (PDL), and cumulative population doublings (CPD) per passage. Pooled batches of hPL from different blood groups were evaluated and compared with Fetal Bovine Serum (FBS) supplement ( n = 4 donors per group). (b) Comparison of human cytokine levels (pg/mL) measured in different hPL batches obtained from blood donors ( n = 3 per blood group). ∗ indicates p
    Figure Legend Snippet: Impact of the use of human platelet lysate (hPL) as a medium supplement for cell culture of UC-MSC. (a) Proliferation kinetics of UC-MSC as measured by population doubling time (hours), population doubling levels (PDL), and cumulative population doublings (CPD) per passage. Pooled batches of hPL from different blood groups were evaluated and compared with Fetal Bovine Serum (FBS) supplement ( n = 4 donors per group). (b) Comparison of human cytokine levels (pg/mL) measured in different hPL batches obtained from blood donors ( n = 3 per blood group). ∗ indicates p

    Techniques Used: Cell Culture

    24) Product Images from "Dual TNF-?/Cyclin D1 Gene Silencing With an Oral Polymeric Microparticle System as a Novel Strategy for the Treatment of Inflammatory Bowel Disease"

    Article Title: Dual TNF-?/Cyclin D1 Gene Silencing With an Oral Polymeric Microparticle System as a Novel Strategy for the Treatment of Inflammatory Bowel Disease

    Journal: Clinical and Translational Gastroenterology

    doi: 10.1038/ctg.2011.1

    Colonic cytokine and chemokine profiles. The cytokine expression profile upon oral delivery of cyclin D1 (CyD1), tumor necrosis factor-α (TNF-α), or a combination of both short interfering RNA (siRNA) encapsulated in nanoparticles-in-microsphere oral system (NiMOS) was determined using a chemiluminescent enzyme-linked immunosorbent assay Q-Plex Mouse Cytokine Screen (Quansys Biosciences). Concentrations of the cytokines ( a ) interferon (IFN)-γ, ( b ) interleukin (IL)-1α, ( c ) IL-1β, ( d ) IL-2, ( e ) IL-5, ( f ) IL-6, ( g ) IL-17 and pro-inflammatory chemokines ( h ) monocyte chemotactic protein (MCP)-1, ( i ) monocyte inflammatory protein (MIP)-1α, and ( j ) granulocyte macrophage colony-stimulating factor (GMCSF) in the large intestine are shown. Administration of CyD1 NiMOS led to a significant reduction in protein expression both time points tested compared with the remaining groups. The effect of combined TNF-α/CyD1 NiMOS and TNF-α was less pronounced, but led to decreased protein levels in comparison with control groups. Values expressed as mean±s.d. ( n =5). Δ P
    Figure Legend Snippet: Colonic cytokine and chemokine profiles. The cytokine expression profile upon oral delivery of cyclin D1 (CyD1), tumor necrosis factor-α (TNF-α), or a combination of both short interfering RNA (siRNA) encapsulated in nanoparticles-in-microsphere oral system (NiMOS) was determined using a chemiluminescent enzyme-linked immunosorbent assay Q-Plex Mouse Cytokine Screen (Quansys Biosciences). Concentrations of the cytokines ( a ) interferon (IFN)-γ, ( b ) interleukin (IL)-1α, ( c ) IL-1β, ( d ) IL-2, ( e ) IL-5, ( f ) IL-6, ( g ) IL-17 and pro-inflammatory chemokines ( h ) monocyte chemotactic protein (MCP)-1, ( i ) monocyte inflammatory protein (MIP)-1α, and ( j ) granulocyte macrophage colony-stimulating factor (GMCSF) in the large intestine are shown. Administration of CyD1 NiMOS led to a significant reduction in protein expression both time points tested compared with the remaining groups. The effect of combined TNF-α/CyD1 NiMOS and TNF-α was less pronounced, but led to decreased protein levels in comparison with control groups. Values expressed as mean±s.d. ( n =5). Δ P

    Techniques Used: Expressing, Small Interfering RNA, Chemiluminescent ELISA

    25) Product Images from "A marine-sourced fucoidan solution inhibits Toll-like-receptor-3-induced cytokine release by human bronchial epithelial cells"

    Article Title: A marine-sourced fucoidan solution inhibits Toll-like-receptor-3-induced cytokine release by human bronchial epithelial cells

    Journal: International Journal of Biological Macromolecules

    doi: 10.1016/j.ijbiomac.2019.02.113

    Effect of the marine-sourced fucoidan solution (MFS) on Poly(I:C)-induced cytokine release (IL-1α, IL-1β, TNF-α, and IL-6) by human bronchial epithelial cells. Bronchial epithelial cells were incubated with the MFS or NaCl 0.9% (control) for 1 h, followed by 24 h in culture medium in the absence or presence of Poly(I:C) 10 μg·mL −1 . The data are expressed as the mean ± SEM of 6 to 12 independent, paired experiments. The significance thresholds were *p
    Figure Legend Snippet: Effect of the marine-sourced fucoidan solution (MFS) on Poly(I:C)-induced cytokine release (IL-1α, IL-1β, TNF-α, and IL-6) by human bronchial epithelial cells. Bronchial epithelial cells were incubated with the MFS or NaCl 0.9% (control) for 1 h, followed by 24 h in culture medium in the absence or presence of Poly(I:C) 10 μg·mL −1 . The data are expressed as the mean ± SEM of 6 to 12 independent, paired experiments. The significance thresholds were *p

    Techniques Used: Incubation

    26) Product Images from "Involvement of Toll-Like Receptors on Helicobacter pylori-Induced Immunity"

    Article Title: Involvement of Toll-Like Receptors on Helicobacter pylori-Induced Immunity

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0104804

    Influence of TLRs on cytokine release by human DCs in response to H. pylori infection. Immature human monocyte-derived DCs were pre-incubated wit TLR-neutralizing antibodies for 1 h and afterwards infected with H. pylori G27 (MOI 5) for 24 h. The release of IL-6, IL-12p70 and IL-10 was determined by ELISA. Data are presented as mean ± SD of six independent experiments. *p≤0.05, **p≤0.005, ***p≤0.0005. Asterisks on top of bars indicate significance relative to non-neutralized, H. pylori -primed control cells.
    Figure Legend Snippet: Influence of TLRs on cytokine release by human DCs in response to H. pylori infection. Immature human monocyte-derived DCs were pre-incubated wit TLR-neutralizing antibodies for 1 h and afterwards infected with H. pylori G27 (MOI 5) for 24 h. The release of IL-6, IL-12p70 and IL-10 was determined by ELISA. Data are presented as mean ± SD of six independent experiments. *p≤0.05, **p≤0.005, ***p≤0.0005. Asterisks on top of bars indicate significance relative to non-neutralized, H. pylori -primed control cells.

    Techniques Used: Infection, Derivative Assay, Incubation, Enzyme-linked Immunosorbent Assay

    27) Product Images from "Mycobacterium abscessus Induces a Limited Pattern of Neutrophil Activation That Promotes Pathogen Survival"

    Article Title: Mycobacterium abscessus Induces a Limited Pattern of Neutrophil Activation That Promotes Pathogen Survival

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0057402

    Secretion of chemokines and cytokines. Neutrophils were stimulated with M. abscessus (Mab; closed bars ), or S. aureus (Sa; open bars ) for 2 and 4 hours, or left unstimulated (NS; hatched bars ) and supernatants were collected. The indicated cytokine and chemokine levels were determined by ELISA. Mean±SEM, n = 7–8; *p
    Figure Legend Snippet: Secretion of chemokines and cytokines. Neutrophils were stimulated with M. abscessus (Mab; closed bars ), or S. aureus (Sa; open bars ) for 2 and 4 hours, or left unstimulated (NS; hatched bars ) and supernatants were collected. The indicated cytokine and chemokine levels were determined by ELISA. Mean±SEM, n = 7–8; *p

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Hierarchical clustering of cytokine and chemokine genes by M. abscessus . Neutrophils were stimulated with M. abscessus (Sm; yellow shading ), or S. aureus (Sa; red shading ) for 2 hours, and gene expression was determined compared to non-stimulated neutrophils (C; blue shading ). Tree spacing indicates linkage distance. Highly expressed ( red ) and low expressed ( green ) genes are indicated from 4 different neutrophil donors.
    Figure Legend Snippet: Hierarchical clustering of cytokine and chemokine genes by M. abscessus . Neutrophils were stimulated with M. abscessus (Sm; yellow shading ), or S. aureus (Sa; red shading ) for 2 hours, and gene expression was determined compared to non-stimulated neutrophils (C; blue shading ). Tree spacing indicates linkage distance. Highly expressed ( red ) and low expressed ( green ) genes are indicated from 4 different neutrophil donors.

    Techniques Used: Expressing

    28) Product Images from "Effect of Heat-Killed Escherichia coli, Lipopolysaccharide, and Muramyl Dipeptide Treatments on the Immune Response Phenotype and Allergy in Neonatal Pigs Sensitized to the Egg White Protein Ovomucoid"

    Article Title: Effect of Heat-Killed Escherichia coli, Lipopolysaccharide, and Muramyl Dipeptide Treatments on the Immune Response Phenotype and Allergy in Neonatal Pigs Sensitized to the Egg White Protein Ovomucoid

    Journal: Clinical and Vaccine Immunology : CVI

    doi: 10.1128/CVI.00555-12

    Ratio of cytokine expression by PHA-P-stimulated BMCs from pigs treated with heat-killed E. coli , LPS, MDP, or PBS (control). Blood was collected from pigs at 45 days of age, postsensitization to ovomucoid. BMCs were isolated using a density gradient of Histopaque (Sigma-Aldrich) and cultured at 2.5 × 10 6 cells/ml for 96 h, stimulated with 10 μg/ml of PHA-P (Sigma-Aldrich). Culture supernatant was collected and stored at −80°C. The concentrations of cytokines were measured using capture enzyme-linked immunosorbent assay (Invitrogen, IL-10; Kingfisher, IL-17; R D Systems, IL-12). Ratios of cytokines were calculated to elucidate possible bias as a result of treatment. There was a trend toward an increased ratio of IL-17 to IL-10 in cells from LPS-treated pigs ( n = 15) compared to those from pigs treated with E. coli ( n = 11). The ratio of IL-17 to IL-12 was greater in supernatant from cells of PBS-treated pigs ( n = 49, pooled across all litters) than in cells from LPS-treated pigs. In cells from LPS-treated pigs, the ratio of IL-10 to IL-12 was less than that in cells from all other treatment groups (unpaired t test, Welch's correction applied when necessary, significance taken at P
    Figure Legend Snippet: Ratio of cytokine expression by PHA-P-stimulated BMCs from pigs treated with heat-killed E. coli , LPS, MDP, or PBS (control). Blood was collected from pigs at 45 days of age, postsensitization to ovomucoid. BMCs were isolated using a density gradient of Histopaque (Sigma-Aldrich) and cultured at 2.5 × 10 6 cells/ml for 96 h, stimulated with 10 μg/ml of PHA-P (Sigma-Aldrich). Culture supernatant was collected and stored at −80°C. The concentrations of cytokines were measured using capture enzyme-linked immunosorbent assay (Invitrogen, IL-10; Kingfisher, IL-17; R D Systems, IL-12). Ratios of cytokines were calculated to elucidate possible bias as a result of treatment. There was a trend toward an increased ratio of IL-17 to IL-10 in cells from LPS-treated pigs ( n = 15) compared to those from pigs treated with E. coli ( n = 11). The ratio of IL-17 to IL-12 was greater in supernatant from cells of PBS-treated pigs ( n = 49, pooled across all litters) than in cells from LPS-treated pigs. In cells from LPS-treated pigs, the ratio of IL-10 to IL-12 was less than that in cells from all other treatment groups (unpaired t test, Welch's correction applied when necessary, significance taken at P

    Techniques Used: Expressing, Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay

    Cytokine expression by PHA-P-stimulated BMCs from piglets treated with heat-killed E. coli , LPS, MDP, or PBS (control). Blood was collected from piglets at 45 days of age, postsensitization to ovomucoid. Blood mononuclear cells (BMCs) from each pig were isolated on a density gradient of Histopaque (Sigma-Aldrich) and cultured at 2.5 × 10 6 cells/ml for 96 h, stimulated with 10 μg/ml of PHA-P (Sigma-Aldrich). Culture supernatant was collected and stored at −80°C. Cytokine concentration was measured using capture enzyme-linked immunosorbent assay (Invitrogen, IL-10 and IL-4; Kingfisher, IL-17; R D Systems, IL-12). The concentration of IL-17 was lower in the supernatant of BMCs from E. coli -treated ( n = 11) pigs than from all others. No difference in the expression of IL-4 was noted from the supernatant of cells from pigs in each treatment group. Cells from piglets treated with MDP ( n = 16) and PBS ( n = 49, pooled across all litters) had an increased expression of IL-10 compared to that in E. coli -treated pigs. Cells from MDP-treated pigs trended toward increased IL-10 production compared to cells from LPS-treated ( n = 15) pigs. The concentration of IL-12 was lower in supernatant from cells of E. coli -treated pigs than in any other treatment group (unpaired t test, Welch's correction applied when necessary, significance taken at P
    Figure Legend Snippet: Cytokine expression by PHA-P-stimulated BMCs from piglets treated with heat-killed E. coli , LPS, MDP, or PBS (control). Blood was collected from piglets at 45 days of age, postsensitization to ovomucoid. Blood mononuclear cells (BMCs) from each pig were isolated on a density gradient of Histopaque (Sigma-Aldrich) and cultured at 2.5 × 10 6 cells/ml for 96 h, stimulated with 10 μg/ml of PHA-P (Sigma-Aldrich). Culture supernatant was collected and stored at −80°C. Cytokine concentration was measured using capture enzyme-linked immunosorbent assay (Invitrogen, IL-10 and IL-4; Kingfisher, IL-17; R D Systems, IL-12). The concentration of IL-17 was lower in the supernatant of BMCs from E. coli -treated ( n = 11) pigs than from all others. No difference in the expression of IL-4 was noted from the supernatant of cells from pigs in each treatment group. Cells from piglets treated with MDP ( n = 16) and PBS ( n = 49, pooled across all litters) had an increased expression of IL-10 compared to that in E. coli -treated pigs. Cells from MDP-treated pigs trended toward increased IL-10 production compared to cells from LPS-treated ( n = 15) pigs. The concentration of IL-12 was lower in supernatant from cells of E. coli -treated pigs than in any other treatment group (unpaired t test, Welch's correction applied when necessary, significance taken at P

    Techniques Used: Expressing, Isolation, Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay

    29) Product Images from "Development and Characterization of Synthetic Glucopyranosyl Lipid Adjuvant System as a Vaccine Adjuvant"

    Article Title: Development and Characterization of Synthetic Glucopyranosyl Lipid Adjuvant System as a Vaccine Adjuvant

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0016333

    Rapid activation of dendritic cells in response to GLA. C57BL/6 mice (n = 6) were immunized 3x, 2 wks apart with ID83 antigen co-administered with 1, 5 or 20 µg of GLA-SE or MPL-SE. Control groups included saline, ID83, and ID83+SE. (A) Mice were bled before (time 0) and 4 h after the first injection. Innate cytokine levels of IL-12p40, TNF, IL-6, MCP-1, CCL5 and CXCL10 were determined by Luminex.
    Figure Legend Snippet: Rapid activation of dendritic cells in response to GLA. C57BL/6 mice (n = 6) were immunized 3x, 2 wks apart with ID83 antigen co-administered with 1, 5 or 20 µg of GLA-SE or MPL-SE. Control groups included saline, ID83, and ID83+SE. (A) Mice were bled before (time 0) and 4 h after the first injection. Innate cytokine levels of IL-12p40, TNF, IL-6, MCP-1, CCL5 and CXCL10 were determined by Luminex.

    Techniques Used: Activation Assay, Mouse Assay, Injection, Luminex

    Dose-dependent activation and maturation of BMDC in response to GLA stimulation. BMDC were stimulated with 0.01–1000 ng/mL of GLA-AF, MPL-AF, LPS or equivalent volumes of AF. (A) IL-12p40, TNF, and IL-6 cytokine levels in culture supernatants after 24 h. (B) Percentage of CD86 + , CD40 + , and MHC Class II + DC cells determined within the CD11c + gate after 48 h of incubation with the TLR4 agonists (C57BL/6, left panels; BALB/c, right panels). (C) DC were pulsed with media or ID83 antigen (50 ng/mL), without or with 1 ng/mL of TLR4 agonist, washed and further incubated with antigen-specific CD4 T cells for 48 h. IFN-γ levels in culture supernatants were measured by ELISA. Data shown are representative of two independent experiments.
    Figure Legend Snippet: Dose-dependent activation and maturation of BMDC in response to GLA stimulation. BMDC were stimulated with 0.01–1000 ng/mL of GLA-AF, MPL-AF, LPS or equivalent volumes of AF. (A) IL-12p40, TNF, and IL-6 cytokine levels in culture supernatants after 24 h. (B) Percentage of CD86 + , CD40 + , and MHC Class II + DC cells determined within the CD11c + gate after 48 h of incubation with the TLR4 agonists (C57BL/6, left panels; BALB/c, right panels). (C) DC were pulsed with media or ID83 antigen (50 ng/mL), without or with 1 ng/mL of TLR4 agonist, washed and further incubated with antigen-specific CD4 T cells for 48 h. IFN-γ levels in culture supernatants were measured by ELISA. Data shown are representative of two independent experiments.

    Techniques Used: Activation Assay, Incubation, Enzyme-linked Immunosorbent Assay

    30) Product Images from "Inflammation of the Fetal Ovine Skin Following in utero Exposure to Ureaplasma parvum"

    Article Title: Inflammation of the Fetal Ovine Skin Following in utero Exposure to Ureaplasma parvum

    Journal: Reproductive Sciences

    doi: 10.1177/1933719111408114

    Quantitative PCR analysis of cytokine/chemokine expression in fetal ovine skin exposed UP in utero for either 7 days or 69 days. Bars represent mean fold change (relative to control) ± SEM. *, statistically significant increase in transcript expression
    Figure Legend Snippet: Quantitative PCR analysis of cytokine/chemokine expression in fetal ovine skin exposed UP in utero for either 7 days or 69 days. Bars represent mean fold change (relative to control) ± SEM. *, statistically significant increase in transcript expression

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, In Utero

    Immunohistochemical analysis of cytokine expression in the fetal skin. Alexa 488 secondary immunofluorescence for IL-1β, IL-8, and TNF-α expression in fetal skin from control, 7-day UP, and 69-day UP exposed animals. UP indicates Ureaplasma
    Figure Legend Snippet: Immunohistochemical analysis of cytokine expression in the fetal skin. Alexa 488 secondary immunofluorescence for IL-1β, IL-8, and TNF-α expression in fetal skin from control, 7-day UP, and 69-day UP exposed animals. UP indicates Ureaplasma

    Techniques Used: Immunohistochemistry, Expressing, Immunofluorescence

    Quantitative PCR analysis of cytokine/chemokine expression in cultured primary keratinocytes exposed to 2 × 107 UP in a time-course experiment at 30 minutes, 1, 2, 4, 6, and 8 hours postexposure. Graph represents mean fold change in infected cells
    Figure Legend Snippet: Quantitative PCR analysis of cytokine/chemokine expression in cultured primary keratinocytes exposed to 2 × 107 UP in a time-course experiment at 30 minutes, 1, 2, 4, 6, and 8 hours postexposure. Graph represents mean fold change in infected cells

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Cell Culture, Infection

    31) Product Images from "The Immune Response to Intrathecal Enzyme Replacement Therapy in Mucopolysaccharidosis I Patients"

    Article Title: The Immune Response to Intrathecal Enzyme Replacement Therapy in Mucopolysaccharidosis I Patients

    Journal: Pediatric research

    doi: 10.1038/pr.2013.158

    Baseline absolute cytokine concentrations in the serum ( a ) and CSF ( b ) of MPS I subjects prior to starting IT rhIDU treatments. T H 1 (cellular immune response) cytokines: IL-1β, IL-2, IL-8, TNFα, IFNγ; T H 2 (humoral immune response)
    Figure Legend Snippet: Baseline absolute cytokine concentrations in the serum ( a ) and CSF ( b ) of MPS I subjects prior to starting IT rhIDU treatments. T H 1 (cellular immune response) cytokines: IL-1β, IL-2, IL-8, TNFα, IFNγ; T H 2 (humoral immune response)

    Techniques Used:

    Cytokine levels in MPS I subjects during the course of IT rhIDU treatments. Absolute concentrations of T H 1 (filled symbols: circles IL-1β; squares IL-2; diamonds IL-8; triangles TNFα; inverted triangles IFNγ) and T H 2 cytokines
    Figure Legend Snippet: Cytokine levels in MPS I subjects during the course of IT rhIDU treatments. Absolute concentrations of T H 1 (filled symbols: circles IL-1β; squares IL-2; diamonds IL-8; triangles TNFα; inverted triangles IFNγ) and T H 2 cytokines

    Techniques Used:

    32) Product Images from "Parasitic Nematode-Induced CD4+Foxp3+T Cells Can Ameliorate Allergic Airway Inflammation"

    Article Title: Parasitic Nematode-Induced CD4+Foxp3+T Cells Can Ameliorate Allergic Airway Inflammation

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0003410

    Cytokine concentrations in BALF and OVA specific lymphocytes isolated from lung draining lymph node. Cytokine concentrations were measured in the BALF samples (A). The lymphocytes were activated by OVA. The wells was incubated with 1 µg/mL of OVA for 16 h at 4°C, and then the lymphocytes isolated from lung draining lymph node (LLN) were added to the well and incubated for three days. After activation, cytokine concentrations in the supernatant were measured using ELISA kits, in accordance with the manufacturer's instructions (B). [OVA-; PBS treated mice, OVA+; allergic airway inflammation-induced mice, IV(inf)+(-); CD4 + Foxp3 + T cell of normal mice adoptive transferred mice, IV(inf)+(+); CD4 + Foxp3 + T cell of T. spiralis-infected mice adoptive transferred mice, * p
    Figure Legend Snippet: Cytokine concentrations in BALF and OVA specific lymphocytes isolated from lung draining lymph node. Cytokine concentrations were measured in the BALF samples (A). The lymphocytes were activated by OVA. The wells was incubated with 1 µg/mL of OVA for 16 h at 4°C, and then the lymphocytes isolated from lung draining lymph node (LLN) were added to the well and incubated for three days. After activation, cytokine concentrations in the supernatant were measured using ELISA kits, in accordance with the manufacturer's instructions (B). [OVA-; PBS treated mice, OVA+; allergic airway inflammation-induced mice, IV(inf)+(-); CD4 + Foxp3 + T cell of normal mice adoptive transferred mice, IV(inf)+(+); CD4 + Foxp3 + T cell of T. spiralis-infected mice adoptive transferred mice, * p

    Techniques Used: Isolation, Incubation, Activation Assay, Enzyme-linked Immunosorbent Assay, Mouse Assay, Infection

    Amelioration of airway inflammation with CD4 + Foxp3 + T cell adoptive transfer during asthma induction (Stage II). The PenH values were determined at baseline and after treatment with increasing doses of aerosolized methacholine (0–50 mg/mL) (A). Relative quantification of MUC2 and MUC5 gene expression in lung after induction of airway inflammation (B). Cytokine concentrations were measured in the BALF samples (C).
    Figure Legend Snippet: Amelioration of airway inflammation with CD4 + Foxp3 + T cell adoptive transfer during asthma induction (Stage II). The PenH values were determined at baseline and after treatment with increasing doses of aerosolized methacholine (0–50 mg/mL) (A). Relative quantification of MUC2 and MUC5 gene expression in lung after induction of airway inflammation (B). Cytokine concentrations were measured in the BALF samples (C).

    Techniques Used: Adoptive Transfer Assay, Expressing

    33) Product Images from "Vitamin D Regulates Cytokine Patterns Secreted by Dendritic Cells to Promote Differentiation of IL-22-Producing T Cells"

    Article Title: Vitamin D Regulates Cytokine Patterns Secreted by Dendritic Cells to Promote Differentiation of IL-22-Producing T Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0130395

    Supernatants of 1,25D diff -DCs promote differentiation of IL-22-secreting T cells. Monocytes were differentiated into DCs with rGM-CSF and rIL-4 in media with 10% FCS in the presence (1,25D diff -DCs) or absence (serum-DCs) of additional 1,25D (10 −8 M). 1,25D diff -DCs and serum-DCs were stimulated with TLR2/1L (1 μg/ml) in fresh media and supernatants were collected after 18–24 hours. Subsequently, TLR2/1-induced 1,25D diff -DC and serum-DC supernatants were added to naïve CD4 + T cells activated with CD3/CD28-coated beads. As a control, naïve CD4 + T cells were incubated with CD3/CD28-coated beads without addition of DC supernatants (beads only). After five days, rIL2 was added to all cultures. On day 12, T cells were re-stimulated with PMA/Ionomycin in fresh media and cytokine secretion evaluated after 18–24 hours. Levels of T cell-derived IL-22, IL-17a and IFN-γ were assessed by ELISA (mean of cytokine levels in ng/ml ± SEM, IL-22 n = 15, IL-17a/IFN-γ n = 13). *p
    Figure Legend Snippet: Supernatants of 1,25D diff -DCs promote differentiation of IL-22-secreting T cells. Monocytes were differentiated into DCs with rGM-CSF and rIL-4 in media with 10% FCS in the presence (1,25D diff -DCs) or absence (serum-DCs) of additional 1,25D (10 −8 M). 1,25D diff -DCs and serum-DCs were stimulated with TLR2/1L (1 μg/ml) in fresh media and supernatants were collected after 18–24 hours. Subsequently, TLR2/1-induced 1,25D diff -DC and serum-DC supernatants were added to naïve CD4 + T cells activated with CD3/CD28-coated beads. As a control, naïve CD4 + T cells were incubated with CD3/CD28-coated beads without addition of DC supernatants (beads only). After five days, rIL2 was added to all cultures. On day 12, T cells were re-stimulated with PMA/Ionomycin in fresh media and cytokine secretion evaluated after 18–24 hours. Levels of T cell-derived IL-22, IL-17a and IFN-γ were assessed by ELISA (mean of cytokine levels in ng/ml ± SEM, IL-22 n = 15, IL-17a/IFN-γ n = 13). *p

    Techniques Used: Incubation, Derivative Assay, Enzyme-linked Immunosorbent Assay

    Effect of 25D on differentiation and/or stimulation of monocyte-derived DCs. Monocytes were differentiated for six days using rGM-CSF and rIL-4 in media with 10% FCS in the presence (25D diff -DCs or 25D diff/stim -DCs) or absence (serum-DCs or 25D stim -DCs) of additional 25D (10 −7 M). Subsequently, 25D-DCs and serum-DCs were stimulated with TLR2/1L (1 μg/ml) or left untreated in the presence (25D diff/stim -DCs or 25D stim -DCs) or absence (serum-DCs or 25D diff -DCs) of additional 25D (10 −7 M), and cultured in fresh media with 10% FCS for 18–24 hours. TNF-α, IL-6, IL-1β, IL-12p40, IL-12p70, IL-23 and IL-10, as well as total and active TGF-β levels in culture supernatants were measured by ELISA or CBA (mean cytokine levels in ng/ml ± SEM, n = 4). Results shown in Fig 1 and Fig. 2 were conducted using cells from the same cell preparations of identical donors. *p
    Figure Legend Snippet: Effect of 25D on differentiation and/or stimulation of monocyte-derived DCs. Monocytes were differentiated for six days using rGM-CSF and rIL-4 in media with 10% FCS in the presence (25D diff -DCs or 25D diff/stim -DCs) or absence (serum-DCs or 25D stim -DCs) of additional 25D (10 −7 M). Subsequently, 25D-DCs and serum-DCs were stimulated with TLR2/1L (1 μg/ml) or left untreated in the presence (25D diff/stim -DCs or 25D stim -DCs) or absence (serum-DCs or 25D diff -DCs) of additional 25D (10 −7 M), and cultured in fresh media with 10% FCS for 18–24 hours. TNF-α, IL-6, IL-1β, IL-12p40, IL-12p70, IL-23 and IL-10, as well as total and active TGF-β levels in culture supernatants were measured by ELISA or CBA (mean cytokine levels in ng/ml ± SEM, n = 4). Results shown in Fig 1 and Fig. 2 were conducted using cells from the same cell preparations of identical donors. *p

    Techniques Used: Derivative Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Crocin Bleaching Assay

    Supernatants of 1,25D diff -DCs promote differentiation of IL-22-expressing T cells. Supernatants of TLR2/1-induced 1,25D diff -DCs and serum-DC were added to naïve CD4 + T cells activated with CD3/CD28-coated beads (as described in Fig 4 ). After five days, rIL2 was added to all cultures. On day 12, T cells were re-stimulated with PMA/Ionomycin for five hours, the last 2.5 hours of culture in the presence of Brefeldin A, in fresh media and intracellular cytokine expression of IL-22, IFN-γ or IL-17a was measured. (A) Dot plots from one representative staining of one donor out of eleven. Upper panel of dot plots shows co-expression of IL-17a and IL-22, lower panel shows co-expression of IFN-γ and IL-22. Numbers above each dot plot indicate frequency of positive cells in each quadrant. (B) Frequency of total IL-22-, IL-17a- and IFN-γ-expressing CD4 + T cells assessed by intracellular cytokine staining (mean percentage of positive cells ± SEM, n = 11). (C) Frequency of IL-22 + /IL-17a + and IL-22 + /IL-17a - or IL-22 + /IFN-γ + and IL-22 + /IFN-γ - CD4 + T cells assessed by intracellular cytokine staining (mean percentage of positive cells ± SEM, n = 11). *p
    Figure Legend Snippet: Supernatants of 1,25D diff -DCs promote differentiation of IL-22-expressing T cells. Supernatants of TLR2/1-induced 1,25D diff -DCs and serum-DC were added to naïve CD4 + T cells activated with CD3/CD28-coated beads (as described in Fig 4 ). After five days, rIL2 was added to all cultures. On day 12, T cells were re-stimulated with PMA/Ionomycin for five hours, the last 2.5 hours of culture in the presence of Brefeldin A, in fresh media and intracellular cytokine expression of IL-22, IFN-γ or IL-17a was measured. (A) Dot plots from one representative staining of one donor out of eleven. Upper panel of dot plots shows co-expression of IL-17a and IL-22, lower panel shows co-expression of IFN-γ and IL-22. Numbers above each dot plot indicate frequency of positive cells in each quadrant. (B) Frequency of total IL-22-, IL-17a- and IFN-γ-expressing CD4 + T cells assessed by intracellular cytokine staining (mean percentage of positive cells ± SEM, n = 11). (C) Frequency of IL-22 + /IL-17a + and IL-22 + /IL-17a - or IL-22 + /IFN-γ + and IL-22 + /IFN-γ - CD4 + T cells assessed by intracellular cytokine staining (mean percentage of positive cells ± SEM, n = 11). *p

    Techniques Used: Expressing, Staining

    Effect of 1,25D on differentiation and/or stimulation of monocyte-derived DCs. Monocytes were differentiated for six days with rGM-CSF and rIL-4 in media with 10% FCS in the presence (1,25D diff -DCs or 1,25D diff/stim -DCs) or absence (serum-DCs or 1,25D stim -DCs) of additional 1,25D (10 −8 M). Subsequently, 1,25D-DCs and serum-DCs were stimulated with TLR2/1L (1 μg/ml) or left untreated in the presence (1,25D diff/stim -DCs or 1,25D stim -DCs) or absence (serum-DCs or 1,25D diff -DCs) of additional 1,25D (10 −8 M) and cultured in fresh media with 10% FCS for 18–24 hours. TNF-α, IL-6, IL-1β, IL-12p40, IL-12p70, IL-23 and IL-10, as well as total and active TGF-β levels in culture supernatants were measured by ELISA or CBA (mean cytokine levels in ng/ml ± SEM, n = 4). Results shown in Fig. 1 and Fig 2 were conducted using cells from the same cell preparations of identical donors. *p
    Figure Legend Snippet: Effect of 1,25D on differentiation and/or stimulation of monocyte-derived DCs. Monocytes were differentiated for six days with rGM-CSF and rIL-4 in media with 10% FCS in the presence (1,25D diff -DCs or 1,25D diff/stim -DCs) or absence (serum-DCs or 1,25D stim -DCs) of additional 1,25D (10 −8 M). Subsequently, 1,25D-DCs and serum-DCs were stimulated with TLR2/1L (1 μg/ml) or left untreated in the presence (1,25D diff/stim -DCs or 1,25D stim -DCs) or absence (serum-DCs or 1,25D diff -DCs) of additional 1,25D (10 −8 M) and cultured in fresh media with 10% FCS for 18–24 hours. TNF-α, IL-6, IL-1β, IL-12p40, IL-12p70, IL-23 and IL-10, as well as total and active TGF-β levels in culture supernatants were measured by ELISA or CBA (mean cytokine levels in ng/ml ± SEM, n = 4). Results shown in Fig. 1 and Fig 2 were conducted using cells from the same cell preparations of identical donors. *p

    Techniques Used: Derivative Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Crocin Bleaching Assay

    Cytokine profiles of 1,25D diff -DCs vs. serum-DCs stimulated with different ligands. Monocytes were differentiated for six days using rGM-CSF and rIL-4 in media with 10% FCS in the presence (1,25D diff -DCs, black bars) or absence (serum-DCs, white bars) of additional 1,25D (10 −8 M). Subsequently, 1,25D diff -DCs and serum-DCs were stimulated with TLR2/1L (1μg/ml), TLR4L (10 ng/ml) or CD40L (5 μg/ml) or left untreated, and cultured in fresh media with 10% FCS for 18–24 hours. TNF-α, IL-6, IL-1β, IL-12p40, IL-12p70, IL-23 and IL-10, as well as total and active TGF-β levels in culture supernatants were measured by ELISA or CBA (mean cytokine levels in ng/ml ± SEM, n of each condition indicated by numbers under the bars). *p
    Figure Legend Snippet: Cytokine profiles of 1,25D diff -DCs vs. serum-DCs stimulated with different ligands. Monocytes were differentiated for six days using rGM-CSF and rIL-4 in media with 10% FCS in the presence (1,25D diff -DCs, black bars) or absence (serum-DCs, white bars) of additional 1,25D (10 −8 M). Subsequently, 1,25D diff -DCs and serum-DCs were stimulated with TLR2/1L (1μg/ml), TLR4L (10 ng/ml) or CD40L (5 μg/ml) or left untreated, and cultured in fresh media with 10% FCS for 18–24 hours. TNF-α, IL-6, IL-1β, IL-12p40, IL-12p70, IL-23 and IL-10, as well as total and active TGF-β levels in culture supernatants were measured by ELISA or CBA (mean cytokine levels in ng/ml ± SEM, n of each condition indicated by numbers under the bars). *p

    Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay, Crocin Bleaching Assay

    1,25D diff -DC-supernatant mediated priming of IL-22-producing T cells is dependent on TNF-α IL-6 and IL-23. Supernatants of TLR2/1-stimulated 1,25D diff -DCs were added to naïve CD4 + T cells activated via CD3/CD28-coated beads (as described in Fig 4 ) in the presence or absence of different monoclonal blocking antibodies as indicated. After five days, rIL2 was added to all cultures. On day 12, T cells were restimulated with PMA/Ionomycin for five hours, the last 2.5 hours of culture in the presence of Brefeldin A, in fresh media and intracellular cytokine expression of IL-22, IFN-γ or IL-17a was measured. Cytokine secretion was evaluated after 18–24 hours without further addition of Brefeldin A. (A) Anti-TNF-α, anti-IL-6R-α (5 μg/ml each) or anti-TGF-β (10 μg/ml). T cell-derived IL-22 assessed by ELISA (mean of cytokine levels in ng/ml ± SEM, n = 5). (B) Anti-TNF-α, anti-IL-6R-α (5 μg/ml each) or anti-TGF-β (10 μg/ml). Frequency of total IL-22-expressing CD4 + T cells assessed by intracellular cytokine staining (mean percentage of positive cells ± SEM, n = 5). (C) Anti-IL-1β or anti-IL-23p40 (5 μg/ml each). Frequency of total IL-22-expressing CD4 + T cells assessed by intracellular cytokine staining (mean percentage of positive cells ± SEM, n = 5). (D) Anti-TNF-α, anti-IL-6R-α or anti-IL-23p40 (5 μg/ml each) blocking antibodies alone or in combination. Dot plots from one representative staining of one donor out of five showing the frequency of IL-22-expressing CD4 + T cells against the sideward-scatter (SSC). Numbers in rectangle gate indicate frequency of positive cells. *p
    Figure Legend Snippet: 1,25D diff -DC-supernatant mediated priming of IL-22-producing T cells is dependent on TNF-α IL-6 and IL-23. Supernatants of TLR2/1-stimulated 1,25D diff -DCs were added to naïve CD4 + T cells activated via CD3/CD28-coated beads (as described in Fig 4 ) in the presence or absence of different monoclonal blocking antibodies as indicated. After five days, rIL2 was added to all cultures. On day 12, T cells were restimulated with PMA/Ionomycin for five hours, the last 2.5 hours of culture in the presence of Brefeldin A, in fresh media and intracellular cytokine expression of IL-22, IFN-γ or IL-17a was measured. Cytokine secretion was evaluated after 18–24 hours without further addition of Brefeldin A. (A) Anti-TNF-α, anti-IL-6R-α (5 μg/ml each) or anti-TGF-β (10 μg/ml). T cell-derived IL-22 assessed by ELISA (mean of cytokine levels in ng/ml ± SEM, n = 5). (B) Anti-TNF-α, anti-IL-6R-α (5 μg/ml each) or anti-TGF-β (10 μg/ml). Frequency of total IL-22-expressing CD4 + T cells assessed by intracellular cytokine staining (mean percentage of positive cells ± SEM, n = 5). (C) Anti-IL-1β or anti-IL-23p40 (5 μg/ml each). Frequency of total IL-22-expressing CD4 + T cells assessed by intracellular cytokine staining (mean percentage of positive cells ± SEM, n = 5). (D) Anti-TNF-α, anti-IL-6R-α or anti-IL-23p40 (5 μg/ml each) blocking antibodies alone or in combination. Dot plots from one representative staining of one donor out of five showing the frequency of IL-22-expressing CD4 + T cells against the sideward-scatter (SSC). Numbers in rectangle gate indicate frequency of positive cells. *p

    Techniques Used: Blocking Assay, Expressing, Derivative Assay, Enzyme-linked Immunosorbent Assay, Staining

    34) Product Images from "Effects of Treatment with the Hypomethylating Agent 5-aza-2′-deoxycytidine in Murine Type II Collagen-Induced Arthritis"

    Article Title: Effects of Treatment with the Hypomethylating Agent 5-aza-2′-deoxycytidine in Murine Type II Collagen-Induced Arthritis

    Journal: Pharmaceuticals

    doi: 10.3390/ph12040174

    Ex vivo evaluation of total anti- type II collagen (CII) IgG ( A ), antigen-specific proliferation ( B ), and cytokine production ( C ) in splenocytes isolated from CIA-affected mice treated in prophylactic regime with vehicle, DAC, or Dex. O.D.—optical density.
    Figure Legend Snippet: Ex vivo evaluation of total anti- type II collagen (CII) IgG ( A ), antigen-specific proliferation ( B ), and cytokine production ( C ) in splenocytes isolated from CIA-affected mice treated in prophylactic regime with vehicle, DAC, or Dex. O.D.—optical density.

    Techniques Used: Ex Vivo, Isolation, Mouse Assay

    35) Product Images from "UDP-glucose 4, 6-dehydratase Activity Plays an Important Role in Maintaining Cell Wall Integrity and Virulence of Candida albicans"

    Article Title: UDP-glucose 4, 6-dehydratase Activity Plays an Important Role in Maintaining Cell Wall Integrity and Virulence of Candida albicans

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1002384

    Reduced virulence is associated with the inability of gal102Δ/Δ to elicit pro-inflammatory cytokine response in the host. (A, B and C) Mice were infected intravenously with WT (SC5314 and DAY286), gal102Δ/Δ, and the GAL102 or the gal102 K159A mutant reintegrants in gal102Δ/Δ strains and were sacrificed at different time points for measurement of serum cytokine amounts. TNFα (A), IFNγ(B) and IL-4 (C) amounts were determined by ELISA. Sera from PBS treated mice were taken as control. Data are represented as means ± standard errors of the means from three separate experiments with sera of five mice. *** p
    Figure Legend Snippet: Reduced virulence is associated with the inability of gal102Δ/Δ to elicit pro-inflammatory cytokine response in the host. (A, B and C) Mice were infected intravenously with WT (SC5314 and DAY286), gal102Δ/Δ, and the GAL102 or the gal102 K159A mutant reintegrants in gal102Δ/Δ strains and were sacrificed at different time points for measurement of serum cytokine amounts. TNFα (A), IFNγ(B) and IL-4 (C) amounts were determined by ELISA. Sera from PBS treated mice were taken as control. Data are represented as means ± standard errors of the means from three separate experiments with sera of five mice. *** p

    Techniques Used: Mouse Assay, Infection, Mutagenesis, Enzyme-linked Immunosorbent Assay

    36) Product Images from "In Vitro Assessment of Cytokine Expression Profile of MCF-7 Cells in Response to hWJ-MSCs Secretome"

    Article Title: In Vitro Assessment of Cytokine Expression Profile of MCF-7 Cells in Response to hWJ-MSCs Secretome

    Journal: Advanced Pharmaceutical Bulletin

    doi: 10.15171/apb.2019.075

    Cytokine expression profile ofhWJ-MSCs and MCF-7 cells
    Figure Legend Snippet: Cytokine expression profile ofhWJ-MSCs and MCF-7 cells

    Techniques Used: Expressing

    Effect ofhWJ-MSCssecretomeon cytokine expression profile of MCF-7 tumor cells
    Figure Legend Snippet: Effect ofhWJ-MSCssecretomeon cytokine expression profile of MCF-7 tumor cells

    Techniques Used: Expressing

    37) Product Images from "Influenza A Viruses Replicate Productively in Mouse Mastocytoma Cells (P815) and Trigger Pro-inflammatory Cytokine and Chemokine Production through TLR3 Signaling Pathway"

    Article Title: Influenza A Viruses Replicate Productively in Mouse Mastocytoma Cells (P815) and Trigger Pro-inflammatory Cytokine and Chemokine Production through TLR3 Signaling Pathway

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2016.02130

    Influenza virus infection causes the release of pro-inflammatory cytokines and chemokines. (A) P815 cells were infected with H1N1, H5N1, and H7N2 at a MOI of 0.1, or mock treated for 24 h. Cell-free supernatants were then analyzed for cytokine and chemokine content using a cytokine array panel. Numbered boxes denote up-regulated expression, with the most dramatic increases annotated in bold. Results shown are representative of two separate experiments. (B) P815 cells were infected with H1N1, H5N1, and H7N2 at a MOI of 0.1, exposed to LE-PolyI:C, or mock treated. At the designated time points, cell supernatants were harvested and the expression of IL-6, IFN-γ, CCL-2, CCL-5, IP-10 TNF-α, IFN-α, and IFN-β was analyzed by ELISA. Graphs shown are mean ± SD of three independent replicates. Asterisks indicate statistically significant increases compared to mock treatment ( ∗ P
    Figure Legend Snippet: Influenza virus infection causes the release of pro-inflammatory cytokines and chemokines. (A) P815 cells were infected with H1N1, H5N1, and H7N2 at a MOI of 0.1, or mock treated for 24 h. Cell-free supernatants were then analyzed for cytokine and chemokine content using a cytokine array panel. Numbered boxes denote up-regulated expression, with the most dramatic increases annotated in bold. Results shown are representative of two separate experiments. (B) P815 cells were infected with H1N1, H5N1, and H7N2 at a MOI of 0.1, exposed to LE-PolyI:C, or mock treated. At the designated time points, cell supernatants were harvested and the expression of IL-6, IFN-γ, CCL-2, CCL-5, IP-10 TNF-α, IFN-α, and IFN-β was analyzed by ELISA. Graphs shown are mean ± SD of three independent replicates. Asterisks indicate statistically significant increases compared to mock treatment ( ∗ P

    Techniques Used: Infection, Expressing, Enzyme-linked Immunosorbent Assay

    38) Product Images from "DNA Vaccines Encoding Interleukin-8 and RANTES Enhance Antigen-Specific Th1-Type CD4+ T-Cell-Mediated Protective Immunity against Herpes Simplex Virus Type 2 In Vivo"

    Article Title: DNA Vaccines Encoding Interleukin-8 and RANTES Enhance Antigen-Specific Th1-Type CD4+ T-Cell-Mediated Protective Immunity against Herpes Simplex Virus Type 2 In Vivo

    Journal: Journal of Virology

    doi:

    Cytokine (A) and chemokine (B) production levels of splenocytes after in vitro gD stimulation in mice coimmunized with chemokine cDNAs. Each group of mice ( n = 2) was immunized with gD DNA vaccines (60 μg per mouse) plus chemokine genes (40 μg per mouse) at 0 and 2 weeks. Two weeks after the last DNA injection, two mice were sacrificed and spleen cells were pooled. Splenocytes were stimulated with 1 μg of gD proteins/ml for 2 days. Samples were assayed in triplicate. Values are means of released cytokine or chemokine concentrations; error bars, standard deviations. Asterisks indicate values that are statistically significant at a P value of
    Figure Legend Snippet: Cytokine (A) and chemokine (B) production levels of splenocytes after in vitro gD stimulation in mice coimmunized with chemokine cDNAs. Each group of mice ( n = 2) was immunized with gD DNA vaccines (60 μg per mouse) plus chemokine genes (40 μg per mouse) at 0 and 2 weeks. Two weeks after the last DNA injection, two mice were sacrificed and spleen cells were pooled. Splenocytes were stimulated with 1 μg of gD proteins/ml for 2 days. Samples were assayed in triplicate. Values are means of released cytokine or chemokine concentrations; error bars, standard deviations. Asterisks indicate values that are statistically significant at a P value of

    Techniques Used: In Vitro, Mouse Assay, Injection

    39) Product Images from "B-1a Lymphocytes Attenuate Insulin Resistance"

    Article Title: B-1a Lymphocytes Attenuate Insulin Resistance

    Journal: Diabetes

    doi: 10.2337/db14-0554

    IL-10 production is impaired in obese mice. A – C : Frequency of GFP + cells within each subpopulation in the PerC, VAT, and spleen from IL-10 EGFP mice fed the NCD or HFD for 9 weeks ( n = 5–6, representative of three experiments). D and E : IL-10 and TNF-α concentration in 48-h PerC and VAT culture supernatants ( n = 6). Body weights ( F ), GTT with AUC analysis ( G ), and fasting insulin ( H ) of HFD B null mice 1 week after receiving PBS, WT B-1a, or IL10 null B-1a cells ( n = 5). I : Serum IgM concentration 1 week after B-1a cell transfer ( n = 5). J : TNF-α production in VAT macrophages. FACS plots are representative of two experiments. SSC-A, side scatter-area. Cytokine concentrations of IL-10 ( K ), TNF-α ( L ), and IL-6 ( M ) in 60-h coculture of WT PerC and VAT macrophages with WT or IL-10 null B-1a cells ( n = 3 per group, two replicates). Values are given as mean ± SEM. * P
    Figure Legend Snippet: IL-10 production is impaired in obese mice. A – C : Frequency of GFP + cells within each subpopulation in the PerC, VAT, and spleen from IL-10 EGFP mice fed the NCD or HFD for 9 weeks ( n = 5–6, representative of three experiments). D and E : IL-10 and TNF-α concentration in 48-h PerC and VAT culture supernatants ( n = 6). Body weights ( F ), GTT with AUC analysis ( G ), and fasting insulin ( H ) of HFD B null mice 1 week after receiving PBS, WT B-1a, or IL10 null B-1a cells ( n = 5). I : Serum IgM concentration 1 week after B-1a cell transfer ( n = 5). J : TNF-α production in VAT macrophages. FACS plots are representative of two experiments. SSC-A, side scatter-area. Cytokine concentrations of IL-10 ( K ), TNF-α ( L ), and IL-6 ( M ) in 60-h coculture of WT PerC and VAT macrophages with WT or IL-10 null B-1a cells ( n = 3 per group, two replicates). Values are given as mean ± SEM. * P

    Techniques Used: Mouse Assay, Concentration Assay, FACS

    Polyclonal IgM, but not monoclonal anti-PC IgM, ameliorates glucose intolerance. Body weights ( A ) and GTT with AUC ( B ) 1 week after receiving PBS, WT B-1a, sIgM null B-1a, WT B-2, or sIgM null B-2 cells ( n = 5–8). Fasting insulin ( C ), IgM concentration in serum ( D ), and VAT lysate ( E ) 1 week after B-1a cell transfer ( n = 4). Body weights ( F ), GTT with AUC ( G ), and fasting insulin ( H ) of HFD B null mice 1 week after receiving PBS, isotype control, polyclonal IgM, or E06 monoclonal anti-PC IgM ( n = 5 for E06 treatment, n = 15 for the rest). I : Anti-PC IgM in serum from NCD and HFD mice ( n = 10). J : Anti-PC IgM in serum from IR and IS obese humans ( n = 32 and 30). OD, optical density. K : Cytokine concentrations in 24-h supernatants from PerC macrophages cultured with the isotype control or polyclonal IgM ( n = 3). MCP-1, monocyte chemoattractant protein-1. Values are given as mean ± SEM. * P
    Figure Legend Snippet: Polyclonal IgM, but not monoclonal anti-PC IgM, ameliorates glucose intolerance. Body weights ( A ) and GTT with AUC ( B ) 1 week after receiving PBS, WT B-1a, sIgM null B-1a, WT B-2, or sIgM null B-2 cells ( n = 5–8). Fasting insulin ( C ), IgM concentration in serum ( D ), and VAT lysate ( E ) 1 week after B-1a cell transfer ( n = 4). Body weights ( F ), GTT with AUC ( G ), and fasting insulin ( H ) of HFD B null mice 1 week after receiving PBS, isotype control, polyclonal IgM, or E06 monoclonal anti-PC IgM ( n = 5 for E06 treatment, n = 15 for the rest). I : Anti-PC IgM in serum from NCD and HFD mice ( n = 10). J : Anti-PC IgM in serum from IR and IS obese humans ( n = 32 and 30). OD, optical density. K : Cytokine concentrations in 24-h supernatants from PerC macrophages cultured with the isotype control or polyclonal IgM ( n = 3). MCP-1, monocyte chemoattractant protein-1. Values are given as mean ± SEM. * P

    Techniques Used: Concentration Assay, Mouse Assay, Cell Culture

    BAFF-deficient (BAFF null ) and anti-BAFF antibody (Ab)–treated obese mice exhibit superior glucose metabolic control compared with WT and B null mice. Body weights ( A ), GTT with AUC ( B ), and ITT ( C ) of HFD WT, BAFF null , and B null mice ( n = 5). GTT with AUC ( D ), body weights ( E ), and fasting insulin ( F ) of HFD WT mice 4 weeks after they received anti-BAFF antibody or isotype control ( n = 5). IgG concentration in serum ( G ) and VAT lysate ( H ) 5 weeks after anti-BAFF antibody treatment ( n = 5). Cytokine concentrations in 24-h cultures of PerC ( I ) or spleen ( J ) cells stimulated with LPS from isotype control and BAFF antibody–treated mice ( n = 5). K : mRNA expression in VAT from isotype control and BAFF antibody–treated mice ( n = 4). Values are given as mean ± SEM. * P
    Figure Legend Snippet: BAFF-deficient (BAFF null ) and anti-BAFF antibody (Ab)–treated obese mice exhibit superior glucose metabolic control compared with WT and B null mice. Body weights ( A ), GTT with AUC ( B ), and ITT ( C ) of HFD WT, BAFF null , and B null mice ( n = 5). GTT with AUC ( D ), body weights ( E ), and fasting insulin ( F ) of HFD WT mice 4 weeks after they received anti-BAFF antibody or isotype control ( n = 5). IgG concentration in serum ( G ) and VAT lysate ( H ) 5 weeks after anti-BAFF antibody treatment ( n = 5). Cytokine concentrations in 24-h cultures of PerC ( I ) or spleen ( J ) cells stimulated with LPS from isotype control and BAFF antibody–treated mice ( n = 5). K : mRNA expression in VAT from isotype control and BAFF antibody–treated mice ( n = 4). Values are given as mean ± SEM. * P

    Techniques Used: Mouse Assay, Concentration Assay, Expressing

    40) Product Images from "Dual R108K and G189D Mutations in the NS1 Protein of A/H1N1 Influenza Virus Counteract Host Innate Immune Responses"

    Article Title: Dual R108K and G189D Mutations in the NS1 Protein of A/H1N1 Influenza Virus Counteract Host Innate Immune Responses

    Journal: Viruses

    doi: 10.3390/v13050905

    Differential expression of cytokines in the lung of mice infected with rXJ49 or XJ49-NS1mut. ( A ) Heat map showing differentially expression of cytokine-encoding genes in infected versus mock-infected lungs of C57BL/6N mice. Samples were analyzed using a 36-Plex mouse ProcartaPlex Panel kit. Expression of each cytokine is presented as log10-fold changes compared with the value in the mock group. ( B ) Expression of IL-6, GM-CSF, CXCL1, CCL2, MIP-1α, MIP-2, IFN-α, and M-CSF in C57BL/6N mice infected with rXJ49, rXJ49-NS1mut, or mock. Average of biological triplicates ± SD are shown. Significance was calculated using one-way ANOVA with multiple comparison tests (*, p
    Figure Legend Snippet: Differential expression of cytokines in the lung of mice infected with rXJ49 or XJ49-NS1mut. ( A ) Heat map showing differentially expression of cytokine-encoding genes in infected versus mock-infected lungs of C57BL/6N mice. Samples were analyzed using a 36-Plex mouse ProcartaPlex Panel kit. Expression of each cytokine is presented as log10-fold changes compared with the value in the mock group. ( B ) Expression of IL-6, GM-CSF, CXCL1, CCL2, MIP-1α, MIP-2, IFN-α, and M-CSF in C57BL/6N mice infected with rXJ49, rXJ49-NS1mut, or mock. Average of biological triplicates ± SD are shown. Significance was calculated using one-way ANOVA with multiple comparison tests (*, p

    Techniques Used: Expressing, Mouse Assay, Infection

    Related Articles

    Staining:

    Article Title: Strong TCRγδ Signaling Prohibits Thymic Development of IL-17A-Secreting γδ T Cells
    Article Snippet: Flow Cytometry For detection of Vγ5Vδ1 and Vγ6Vδ1, cells were pre-stained with GL3 followed by 17D1. .. For i.c. cytokine staining (eBioscience), cells were stimulated with 50 ng/ml phorbol 12-myristate 13-acetate (PMA; Sigma) and 1 μg/ml ionomycin (Sigma) for 4 hr at 37°C. .. Acquisition was performed with an LSR-II or a Canto II (BD).

    Article Title: Sequential transcriptional changes dictate safe and effective antigen-specific immunotherapy
    Article Snippet: Cells were stained with Vβ8-FITC (clone KJ16-133, diluted 1:100) or with fixable viability dye eFluor780 (1:1,000) before surface staining with CD4-Alexa700 (GK1.5, 1:100) and fixation using IC fixation buffer (all from eBioscience). .. Antibodies for intracellular cytokine staining were diluted in permeabilization buffer; IL-10-allophycocyanin (APC) (JES5-16E3, 1:200), IFN-γ-PerCP-Cy5.5 (XMG1.2, 1:200) (both from eBioscience). .. For IL-21 staining, 4 μg ml−1 recombinant mouse IL-21R Fc chimera (R & D Systems) was used, followed by anti-human immunoglobulin G-PE (Fc γ-specific, 1:250) (eBioscience).

    Article Title: Fucoidan from Macrocystis pyrifera Has Powerful Immune-Modulatory Effects Compared to Three Other Fucoidans
    Article Snippet: Ex Vivo T Cell Stimulation and Intracellular Cytokine Staining Singles cells prepared from spleens were stimulated in vitro for 4 h with phorbol 12-myristate 13-acetate (50 ng/mL) and ionomycin (1 μM; both from Calbiochem, Billerica, MA, USA), with the addition of monensin solution (Biolegend) during the final 2 h. Cells were then stained for surface markers. .. For intracellular cytokine staining, cells were stained for surface molecules first, then fixed and permeabilized with Cytofix/Cytoperm buffer (eBioscience) and subsequently incubated with anti-cytokine antibodies in Perm/Wash buffer (eBioscience) for 30 min. Control staining with isotype control IgGs was performed in all experiments. .. ELISA IL-6, IL-8 and TNF-α protein levels in neutrophil cultured medium and IL-6, IL-12p70, IL-23 (p19/p40) and TNF-α concentrations in the sera were measured in triplicate using standard ELISA kits (Biolegend).

    Neutralization:

    Article Title: Toll-like receptor-4 mediates cigarette smoke-induced cytokine production by human macrophages
    Article Snippet: After 24 hours, the cells were permeabilized in Cytofix/Cytoperm™ solution and stained for intracellular cytokine expression (all from Pharmingen, San Diego, CA, USA). .. Anti-TLR neutralization of cytokine production Cells were incubated with anti-human TLR2 (clone TL2.1) or mouse IgG2a isotype control (20 μg/ml), for 30 min at room temperature or with anti-human TLR4 (clone HTA125) or mouse IgG2a isotype control (20 μg/ml), (all from eBioscience, CA, USA) for one hr at 37°C. .. Hereafter, cells were stimulated with different concentrations of CS medium or LPS or PMA/ionomycin (Sigma) and incubated overnight.

    Incubation:

    Article Title: Toll-like receptor-4 mediates cigarette smoke-induced cytokine production by human macrophages
    Article Snippet: After 24 hours, the cells were permeabilized in Cytofix/Cytoperm™ solution and stained for intracellular cytokine expression (all from Pharmingen, San Diego, CA, USA). .. Anti-TLR neutralization of cytokine production Cells were incubated with anti-human TLR2 (clone TL2.1) or mouse IgG2a isotype control (20 μg/ml), for 30 min at room temperature or with anti-human TLR4 (clone HTA125) or mouse IgG2a isotype control (20 μg/ml), (all from eBioscience, CA, USA) for one hr at 37°C. .. Hereafter, cells were stimulated with different concentrations of CS medium or LPS or PMA/ionomycin (Sigma) and incubated overnight.

    Article Title: Fucoidan from Macrocystis pyrifera Has Powerful Immune-Modulatory Effects Compared to Three Other Fucoidans
    Article Snippet: Ex Vivo T Cell Stimulation and Intracellular Cytokine Staining Singles cells prepared from spleens were stimulated in vitro for 4 h with phorbol 12-myristate 13-acetate (50 ng/mL) and ionomycin (1 μM; both from Calbiochem, Billerica, MA, USA), with the addition of monensin solution (Biolegend) during the final 2 h. Cells were then stained for surface markers. .. For intracellular cytokine staining, cells were stained for surface molecules first, then fixed and permeabilized with Cytofix/Cytoperm buffer (eBioscience) and subsequently incubated with anti-cytokine antibodies in Perm/Wash buffer (eBioscience) for 30 min. Control staining with isotype control IgGs was performed in all experiments. .. ELISA IL-6, IL-8 and TNF-α protein levels in neutrophil cultured medium and IL-6, IL-12p70, IL-23 (p19/p40) and TNF-α concentrations in the sera were measured in triplicate using standard ELISA kits (Biolegend).

    Infection:

    Article Title: Live Attenuated Leishmania donovani Centrin Knock Out Parasites Generate Non-inferior Protective Immune Response in Aged Mice against Visceral Leishmaniasis
    Article Snippet: For cytokine and NO measurements, DCs were infected with parasites and stimulated with LPS (1μg/ml) for 24h. .. Culture supernatants were collected at 24h post infection to evaluate cytokine (IL-12, TNF and IL-6) production with the use of sandwich ELISA kit (ebioscience) and nitric oxide production by the Griess assay. .. Infection of mice, parasite burden estimation and parasitized splenic dendritic cell isolation The young and aged mice were infected via tail vein with 3 X 106 stationary phase red fluorescent LdWT RFP , or LdCen-/-m-cherry promastigotes.

    Article Title: Modulation of the immune response by Fonsecaea pedrosoi morphotypes in the course of experimental chromoblastomycosis and their role on inflammatory response chronicity
    Article Snippet: Measurement of nitric oxide (NO) concentration and cytokine production assay NO2 concentration in culture supernatants was used as an indicator of NO generation and it was measured using Griess reagent as described previously [ ]. .. Cytokine production was measured from homogenized tissue obtained from infected animals as well as cell culture supernatants by Elisa, following manufactory instructions: interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor (TNF), monocyte chemoattractant protein-1 (MCP-1/Ccl2) (eBioscience) and Interleukin-12 (IL-12) (BD Biosciences). ..

    Sandwich ELISA:

    Article Title: Live Attenuated Leishmania donovani Centrin Knock Out Parasites Generate Non-inferior Protective Immune Response in Aged Mice against Visceral Leishmaniasis
    Article Snippet: For cytokine and NO measurements, DCs were infected with parasites and stimulated with LPS (1μg/ml) for 24h. .. Culture supernatants were collected at 24h post infection to evaluate cytokine (IL-12, TNF and IL-6) production with the use of sandwich ELISA kit (ebioscience) and nitric oxide production by the Griess assay. .. Infection of mice, parasite burden estimation and parasitized splenic dendritic cell isolation The young and aged mice were infected via tail vein with 3 X 106 stationary phase red fluorescent LdWT RFP , or LdCen-/-m-cherry promastigotes.

    Griess Assay:

    Article Title: Live Attenuated Leishmania donovani Centrin Knock Out Parasites Generate Non-inferior Protective Immune Response in Aged Mice against Visceral Leishmaniasis
    Article Snippet: For cytokine and NO measurements, DCs were infected with parasites and stimulated with LPS (1μg/ml) for 24h. .. Culture supernatants were collected at 24h post infection to evaluate cytokine (IL-12, TNF and IL-6) production with the use of sandwich ELISA kit (ebioscience) and nitric oxide production by the Griess assay. .. Infection of mice, parasite burden estimation and parasitized splenic dendritic cell isolation The young and aged mice were infected via tail vein with 3 X 106 stationary phase red fluorescent LdWT RFP , or LdCen-/-m-cherry promastigotes.

    Functional Assay:

    Article Title: T Cell Costimulation by CD6 Is Dependent on Bivalent Binding of a GADS/SLP-76 Complex
    Article Snippet: A Jurkat T cell line (expressing the 1G4 TCR; provided by Oreste Acuto, University of Oxford) ( ) or primary T cell blasts were transduced with lentivirus , and when necessary, Jurkat cells were selected for expression by using magnetic beads coated with the T12.1 CD6 MAb (ATCC). .. For functional assays, 96-well round-bottom plates were coated with CD3 (UCHT1; eBioscience) and/or human (T12.1) or rat (OX52; European Collection of Authenticated Cell Cultures [ECACC]) CD6 MAbs. .. Jurkat or activated primary CD4+ T cells (5 × 105 cells in 200 μl) expressing CD6 or mutants were added, and the mixture was incubated for 18 h at 37°C.

    Cell Culture:

    Article Title: The VEGF-Receptor Inhibitor Axitinib Impairs Dendritic Cell Phenotype and Function
    Article Snippet: Immunostaining MoDCs were stained using commercially available mAbs from BD Biosciences, Dako Diagnostika, Immunotech, R & D Systems and eBioscience. .. Determination of cytokine production For cytokine analysis in cell culture supernatants, FlowCytomix Multiple Analyte Detection commercial assay kits were used (eBioscience) and Flow Cytomix Pro Software. .. In vitro migration assay A total of 1x105 cells were seeded into a transwell chamber (8 μm; BD Falcon) in a 24-well plate, and migration to CCL19/MIP-3β was analyzed after 4 h by counting gated DCs for 1 minute in a FACS cytometer.

    Article Title: Modulation of the immune response by Fonsecaea pedrosoi morphotypes in the course of experimental chromoblastomycosis and their role on inflammatory response chronicity
    Article Snippet: Measurement of nitric oxide (NO) concentration and cytokine production assay NO2 concentration in culture supernatants was used as an indicator of NO generation and it was measured using Griess reagent as described previously [ ]. .. Cytokine production was measured from homogenized tissue obtained from infected animals as well as cell culture supernatants by Elisa, following manufactory instructions: interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor (TNF), monocyte chemoattractant protein-1 (MCP-1/Ccl2) (eBioscience) and Interleukin-12 (IL-12) (BD Biosciences). ..

    Flow Cytometry:

    Article Title: The VEGF-Receptor Inhibitor Axitinib Impairs Dendritic Cell Phenotype and Function
    Article Snippet: Immunostaining MoDCs were stained using commercially available mAbs from BD Biosciences, Dako Diagnostika, Immunotech, R & D Systems and eBioscience. .. Determination of cytokine production For cytokine analysis in cell culture supernatants, FlowCytomix Multiple Analyte Detection commercial assay kits were used (eBioscience) and Flow Cytomix Pro Software. .. In vitro migration assay A total of 1x105 cells were seeded into a transwell chamber (8 μm; BD Falcon) in a 24-well plate, and migration to CCL19/MIP-3β was analyzed after 4 h by counting gated DCs for 1 minute in a FACS cytometer.

    Software:

    Article Title: The VEGF-Receptor Inhibitor Axitinib Impairs Dendritic Cell Phenotype and Function
    Article Snippet: Immunostaining MoDCs were stained using commercially available mAbs from BD Biosciences, Dako Diagnostika, Immunotech, R & D Systems and eBioscience. .. Determination of cytokine production For cytokine analysis in cell culture supernatants, FlowCytomix Multiple Analyte Detection commercial assay kits were used (eBioscience) and Flow Cytomix Pro Software. .. In vitro migration assay A total of 1x105 cells were seeded into a transwell chamber (8 μm; BD Falcon) in a 24-well plate, and migration to CCL19/MIP-3β was analyzed after 4 h by counting gated DCs for 1 minute in a FACS cytometer.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Modulation of the immune response by Fonsecaea pedrosoi morphotypes in the course of experimental chromoblastomycosis and their role on inflammatory response chronicity
    Article Snippet: Measurement of nitric oxide (NO) concentration and cytokine production assay NO2 concentration in culture supernatants was used as an indicator of NO generation and it was measured using Griess reagent as described previously [ ]. .. Cytokine production was measured from homogenized tissue obtained from infected animals as well as cell culture supernatants by Elisa, following manufactory instructions: interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor (TNF), monocyte chemoattractant protein-1 (MCP-1/Ccl2) (eBioscience) and Interleukin-12 (IL-12) (BD Biosciences). ..

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  • 98
    Thermo Fisher cytokines
    Coexpression Pattern of Th2 Suppressor <t>Cytokines</t> with Cyp11a1 at Single-Cell Level (A) Gating strategy used to purify subpopulations of Th2 cells based on their cell generation and IL-13-GFP expression. Naive Th cells obtained from spleen of IL-13-GFP reporter mice were stained with CellTrace Violet dye and polarized for Th2 (72–96 hr). The fourth-generation cells that expressed IL-13-GFP (G4P) and second-generation cells that did not express IL-13-GFP (G2N) were FACS sorted and used for single-cell gene expression analysis by qPCR and for single-cell RNA sequencing. (B) Spearman correlation coefficient (r) of Th2-associated genes with Cyp11a1 at single-cell level. Th2 cells from the G2N and G4P groups, as shown in (A) were FACS sorted as single cells. mRNA expression of different protein factors in a single cell were analyzed by single-cell qPCR. (C) Hierarchical clustering of normalized mRNA expression data in single Th2 cell by qPCR for 73 selected genes. The clustering heatmap depicts patterns of coexpression among genes, where the green and red colors indicate the strength and direction of the gene-gene correlation (red meaning higher degrees of similarity, and green lower degrees of similarity). Hierarchical clustering was applied to group genes based on the similarity of their expression profile calculated by Spearman correlation across this data set. The gene cluster containing Cyp11a1 is boxed white, which contains Cyp11a1, IL-10, and TGF-β1.
    Cytokines, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cytokines/product/Thermo Fisher
    Average 98 stars, based on 1 article reviews
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    cytokines - by Bioz Stars, 2021-06
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    97
    Thermo Fisher cytokine analysis
    Peramivir inhibits <t>cytokine</t> release in LPS-induced hPBMCs from a health donor. TNF-α concentration was elevated by LPS stimulation. a and b Peramivir reduced TNF-α and IL-10 release in a time (6 and 12 h)- and dose (2.5, 5, 10 μM)-dependent manner. Peramivir showed no toxicity toward hPBMCs. * P
    Cytokine Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cytokine analysis/product/Thermo Fisher
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    99
    Thermo Fisher b invitrogen human cytokine magnetic 30 plex panel
    Mean difference dot plots of IL-10 and IL-2 cytokines for each kit tested. A) IL-10 and B) IL-2 disagreement plots show the difference between the duplicates against the geometric mean of both values of a sample tested with eBioscience® FlowCytomix™ (FlowCytomix), Millipore™ MILLIPLEX® MAP Plex Kit (Millipore), Bio-Rad® Bio-Plex Pro™ Human <t>Cytokine</t> Plex Assay (Bio-Rad), <t>Invitrogen™</t> Human Cytokine Magnetic 30-Plex Panel (INV-MAG) and BD™ Cytometric Bead Array Human Enhanced Sensitivity kit (BD CBA). The middle line is the mean difference and the two extreme lines are the limits of agreement (values are truncated to the first decimal) calculated by Bland-Altman test.
    B Invitrogen Human Cytokine Magnetic 30 Plex Panel, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher cytokine analysis cell free supernatants
    Supernatants from co-cultures of M-CSF MΦ and HCMV-infected RPE cells inhibit viral spread. Cell-free supernatants from uninfected or HCMV-infected RPE (MOI 12) or M-CSF-MΦ (MOI 3) mono-cultures and RPE/M-CSF-MΦ co-cultures were harvested 2 dpi and transferred onto fresh RPE cells infected with HCMV-GFP at MOI 0.1. (a) 8 dpi virus plaque formation was analyzed by fluorescence microscopy (scale bar = 1 mm) and (b) the size of individual plaques was determined. Cell-free supernatants prepared as described above were analyzed for (c) secreted IFN-α and (d) IFN-I activity using an ELISA method or an Mx2-Luc reporter cell line, respectively. (e) RPE cells were infected with HCMV-GFP at MOI 0.1 and incubated with recombinant (rec.) IFN-α2b, IFN-γ, or TNF-α for 8 d. Then the plaque size was analyzed. (f) Cell-free supernatants prepared as described above were analyzed for different secreted proteins using a bead-based <t>cytokine</t> array. Data visualize log (2) values of the measured cytokine concentrations in [pg/ml]. Mean size of (b) 67–371 plaques using 6 different donors or (e) 49–100 plaques from 3 independent experiments. Mean ±SEM of (c/d) 6 different donors from 3 independent experiments or data of (f) 3 different donors from 2 independent experiments. SN = supernatant, uninf. = uninfected, *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001 (one-tailed Mann-Whitney test).
    Cytokine Analysis Cell Free Supernatants, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Coexpression Pattern of Th2 Suppressor Cytokines with Cyp11a1 at Single-Cell Level (A) Gating strategy used to purify subpopulations of Th2 cells based on their cell generation and IL-13-GFP expression. Naive Th cells obtained from spleen of IL-13-GFP reporter mice were stained with CellTrace Violet dye and polarized for Th2 (72–96 hr). The fourth-generation cells that expressed IL-13-GFP (G4P) and second-generation cells that did not express IL-13-GFP (G2N) were FACS sorted and used for single-cell gene expression analysis by qPCR and for single-cell RNA sequencing. (B) Spearman correlation coefficient (r) of Th2-associated genes with Cyp11a1 at single-cell level. Th2 cells from the G2N and G4P groups, as shown in (A) were FACS sorted as single cells. mRNA expression of different protein factors in a single cell were analyzed by single-cell qPCR. (C) Hierarchical clustering of normalized mRNA expression data in single Th2 cell by qPCR for 73 selected genes. The clustering heatmap depicts patterns of coexpression among genes, where the green and red colors indicate the strength and direction of the gene-gene correlation (red meaning higher degrees of similarity, and green lower degrees of similarity). Hierarchical clustering was applied to group genes based on the similarity of their expression profile calculated by Spearman correlation across this data set. The gene cluster containing Cyp11a1 is boxed white, which contains Cyp11a1, IL-10, and TGF-β1.

    Journal: Cell Reports

    Article Title: Single-Cell RNA Sequencing Reveals T Helper Cells Synthesizing Steroids De Novo to Contribute to Immune Homeostasis

    doi: 10.1016/j.celrep.2014.04.011

    Figure Lengend Snippet: Coexpression Pattern of Th2 Suppressor Cytokines with Cyp11a1 at Single-Cell Level (A) Gating strategy used to purify subpopulations of Th2 cells based on their cell generation and IL-13-GFP expression. Naive Th cells obtained from spleen of IL-13-GFP reporter mice were stained with CellTrace Violet dye and polarized for Th2 (72–96 hr). The fourth-generation cells that expressed IL-13-GFP (G4P) and second-generation cells that did not express IL-13-GFP (G2N) were FACS sorted and used for single-cell gene expression analysis by qPCR and for single-cell RNA sequencing. (B) Spearman correlation coefficient (r) of Th2-associated genes with Cyp11a1 at single-cell level. Th2 cells from the G2N and G4P groups, as shown in (A) were FACS sorted as single cells. mRNA expression of different protein factors in a single cell were analyzed by single-cell qPCR. (C) Hierarchical clustering of normalized mRNA expression data in single Th2 cell by qPCR for 73 selected genes. The clustering heatmap depicts patterns of coexpression among genes, where the green and red colors indicate the strength and direction of the gene-gene correlation (red meaning higher degrees of similarity, and green lower degrees of similarity). Hierarchical clustering was applied to group genes based on the similarity of their expression profile calculated by Spearman correlation across this data set. The gene cluster containing Cyp11a1 is boxed white, which contains Cyp11a1, IL-10, and TGF-β1.

    Article Snippet: In vitro Th cell experiments: staining was performed following eBioscience intracellular staining protocol for cytokines and nuclear staining/transcription factor staining protocol for different transcription factors (GATA3, FOXP3) and Cyp11a1, using eBioscience reagents and kits.

    Techniques: Expressing, Mouse Assay, Staining, FACS, Real-time Polymerase Chain Reaction, RNA Sequencing Assay

    Peramivir inhibits cytokine release in LPS-induced hPBMCs from a health donor. TNF-α concentration was elevated by LPS stimulation. a and b Peramivir reduced TNF-α and IL-10 release in a time (6 and 12 h)- and dose (2.5, 5, 10 μM)-dependent manner. Peramivir showed no toxicity toward hPBMCs. * P

    Journal: bioRxiv

    Article Title: Peramivir, an anti-influenza virus drug, exhibits potential anti-cytokine storm effects

    doi: 10.1101/2020.07.13.201806

    Figure Lengend Snippet: Peramivir inhibits cytokine release in LPS-induced hPBMCs from a health donor. TNF-α concentration was elevated by LPS stimulation. a and b Peramivir reduced TNF-α and IL-10 release in a time (6 and 12 h)- and dose (2.5, 5, 10 μM)-dependent manner. Peramivir showed no toxicity toward hPBMCs. * P

    Article Snippet: 3 × 10 cells were seeded in 96-well plates and incubated for 24 h at 37°C in a humidified atmosphere containing 5% CO2. hPBMCs were pretreated with peramivir at the concentrations of 2.5, 5 and 10 μM at 1 h before LPS (100 ng/ml) stimulation, and the supernatants were harvested at 6 or 12 h after LPS stimulation for cytokine analysis.

    Techniques: Concentration Assay

    Mean difference dot plots of IL-10 and IL-2 cytokines for each kit tested. A) IL-10 and B) IL-2 disagreement plots show the difference between the duplicates against the geometric mean of both values of a sample tested with eBioscience® FlowCytomix™ (FlowCytomix), Millipore™ MILLIPLEX® MAP Plex Kit (Millipore), Bio-Rad® Bio-Plex Pro™ Human Cytokine Plex Assay (Bio-Rad), Invitrogen™ Human Cytokine Magnetic 30-Plex Panel (INV-MAG) and BD™ Cytometric Bead Array Human Enhanced Sensitivity kit (BD CBA). The middle line is the mean difference and the two extreme lines are the limits of agreement (values are truncated to the first decimal) calculated by Bland-Altman test.

    Journal: PLoS ONE

    Article Title: Performance of Multiplex Commercial Kits to Quantify Cytokine and Chemokine Responses in Culture Supernatants from Plasmodium falciparum Stimulations

    doi: 10.1371/journal.pone.0052587

    Figure Lengend Snippet: Mean difference dot plots of IL-10 and IL-2 cytokines for each kit tested. A) IL-10 and B) IL-2 disagreement plots show the difference between the duplicates against the geometric mean of both values of a sample tested with eBioscience® FlowCytomix™ (FlowCytomix), Millipore™ MILLIPLEX® MAP Plex Kit (Millipore), Bio-Rad® Bio-Plex Pro™ Human Cytokine Plex Assay (Bio-Rad), Invitrogen™ Human Cytokine Magnetic 30-Plex Panel (INV-MAG) and BD™ Cytometric Bead Array Human Enhanced Sensitivity kit (BD CBA). The middle line is the mean difference and the two extreme lines are the limits of agreement (values are truncated to the first decimal) calculated by Bland-Altman test.

    Article Snippet: Disagreement plots show the difference between the duplicates against the geometric mean of both values of a sample tested with A) Human Cytokine 25-Plex panel from Invitrogen™ (non-magnetic beads) and B) Invitrogen™ Human Cytokine Magnetic 30-Plex Panel (INV-MAG).

    Techniques: Plex Assay, Crocin Bleaching Assay

    Supernatants from co-cultures of M-CSF MΦ and HCMV-infected RPE cells inhibit viral spread. Cell-free supernatants from uninfected or HCMV-infected RPE (MOI 12) or M-CSF-MΦ (MOI 3) mono-cultures and RPE/M-CSF-MΦ co-cultures were harvested 2 dpi and transferred onto fresh RPE cells infected with HCMV-GFP at MOI 0.1. (a) 8 dpi virus plaque formation was analyzed by fluorescence microscopy (scale bar = 1 mm) and (b) the size of individual plaques was determined. Cell-free supernatants prepared as described above were analyzed for (c) secreted IFN-α and (d) IFN-I activity using an ELISA method or an Mx2-Luc reporter cell line, respectively. (e) RPE cells were infected with HCMV-GFP at MOI 0.1 and incubated with recombinant (rec.) IFN-α2b, IFN-γ, or TNF-α for 8 d. Then the plaque size was analyzed. (f) Cell-free supernatants prepared as described above were analyzed for different secreted proteins using a bead-based cytokine array. Data visualize log (2) values of the measured cytokine concentrations in [pg/ml]. Mean size of (b) 67–371 plaques using 6 different donors or (e) 49–100 plaques from 3 independent experiments. Mean ±SEM of (c/d) 6 different donors from 3 independent experiments or data of (f) 3 different donors from 2 independent experiments. SN = supernatant, uninf. = uninfected, *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001 (one-tailed Mann-Whitney test).

    Journal: Virulence

    Article Title: Human monocyte-derived macrophages inhibit HCMV spread independent of classical antiviral cytokines

    doi: 10.1080/21505594.2018.1535785

    Figure Lengend Snippet: Supernatants from co-cultures of M-CSF MΦ and HCMV-infected RPE cells inhibit viral spread. Cell-free supernatants from uninfected or HCMV-infected RPE (MOI 12) or M-CSF-MΦ (MOI 3) mono-cultures and RPE/M-CSF-MΦ co-cultures were harvested 2 dpi and transferred onto fresh RPE cells infected with HCMV-GFP at MOI 0.1. (a) 8 dpi virus plaque formation was analyzed by fluorescence microscopy (scale bar = 1 mm) and (b) the size of individual plaques was determined. Cell-free supernatants prepared as described above were analyzed for (c) secreted IFN-α and (d) IFN-I activity using an ELISA method or an Mx2-Luc reporter cell line, respectively. (e) RPE cells were infected with HCMV-GFP at MOI 0.1 and incubated with recombinant (rec.) IFN-α2b, IFN-γ, or TNF-α for 8 d. Then the plaque size was analyzed. (f) Cell-free supernatants prepared as described above were analyzed for different secreted proteins using a bead-based cytokine array. Data visualize log (2) values of the measured cytokine concentrations in [pg/ml]. Mean size of (b) 67–371 plaques using 6 different donors or (e) 49–100 plaques from 3 independent experiments. Mean ±SEM of (c/d) 6 different donors from 3 independent experiments or data of (f) 3 different donors from 2 independent experiments. SN = supernatant, uninf. = uninfected, *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001 (one-tailed Mann-Whitney test).

    Article Snippet: Cytokine analysis Cell-free supernatants were analyzed by using Human IFN-alpha Platinum ELISA (eBioscience, BMS216TEN) or Human Inflammation 20plex FlowCytomix Multiplex Kit (eBioscience, BMS819FFRTU) according to the manufacturer’s instructions.

    Techniques: Infection, Fluorescence, Microscopy, Activity Assay, Enzyme-linked Immunosorbent Assay, Incubation, Recombinant, One-tailed Test, MANN-WHITNEY