Structured Review

Millipore phytohemagglutinin pha
Cytokine production by CD4 and CD8 T cells, by CD4 and CD8 T-cell types, and by fold increase upon stimulation. <t>PBMCs</t> were stimulated by phytohemagglutinin or by no mitogen stimulant for 24 hours. Cells were harvested and stained with fluorochrome conjugated CD3, CD4, CD8, IFN-γ, IL-2 or TNF-α mAbs. PBMCs were gated by forward and side scatter and the CD3 + T lymphocyte population was identified (A). IFN-γ, IL-2 or TNF-α expression in CD4 + T cells (B) or CD8+ T cells (C) were then examined. Figures 2D and 2E show fold increase of IFN-γ, Figures 2F and 2G fold increases of IL-2, and Figures 2H and 2I fold increases of TNF-α production in response to <t>PHA,</t> compared to cells cultured with no mitogen stimulant. Symbols represents the data of each individual case patient (circles), HIV+ control (up-pointing triangles), and healthy control (down-pointing triangles). Long horizontal lines show the medians, and short lines the interquartile ranges of each group. Indicated p values for comparisons are by Kruskal-wallis one-way analysis of variance by rank test.
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1) Product Images from "Cellular Immune Responses in HIV-Negative Immunodeficiency with Anti-Interferon-γ Antibodies and Opportunistic Intracellular Microorganisms"

Article Title: Cellular Immune Responses in HIV-Negative Immunodeficiency with Anti-Interferon-γ Antibodies and Opportunistic Intracellular Microorganisms

Journal: PLoS ONE

doi: 10.1371/journal.pone.0110276

Cytokine production by CD4 and CD8 T cells, by CD4 and CD8 T-cell types, and by fold increase upon stimulation. PBMCs were stimulated by phytohemagglutinin or by no mitogen stimulant for 24 hours. Cells were harvested and stained with fluorochrome conjugated CD3, CD4, CD8, IFN-γ, IL-2 or TNF-α mAbs. PBMCs were gated by forward and side scatter and the CD3 + T lymphocyte population was identified (A). IFN-γ, IL-2 or TNF-α expression in CD4 + T cells (B) or CD8+ T cells (C) were then examined. Figures 2D and 2E show fold increase of IFN-γ, Figures 2F and 2G fold increases of IL-2, and Figures 2H and 2I fold increases of TNF-α production in response to PHA, compared to cells cultured with no mitogen stimulant. Symbols represents the data of each individual case patient (circles), HIV+ control (up-pointing triangles), and healthy control (down-pointing triangles). Long horizontal lines show the medians, and short lines the interquartile ranges of each group. Indicated p values for comparisons are by Kruskal-wallis one-way analysis of variance by rank test.
Figure Legend Snippet: Cytokine production by CD4 and CD8 T cells, by CD4 and CD8 T-cell types, and by fold increase upon stimulation. PBMCs were stimulated by phytohemagglutinin or by no mitogen stimulant for 24 hours. Cells were harvested and stained with fluorochrome conjugated CD3, CD4, CD8, IFN-γ, IL-2 or TNF-α mAbs. PBMCs were gated by forward and side scatter and the CD3 + T lymphocyte population was identified (A). IFN-γ, IL-2 or TNF-α expression in CD4 + T cells (B) or CD8+ T cells (C) were then examined. Figures 2D and 2E show fold increase of IFN-γ, Figures 2F and 2G fold increases of IL-2, and Figures 2H and 2I fold increases of TNF-α production in response to PHA, compared to cells cultured with no mitogen stimulant. Symbols represents the data of each individual case patient (circles), HIV+ control (up-pointing triangles), and healthy control (down-pointing triangles). Long horizontal lines show the medians, and short lines the interquartile ranges of each group. Indicated p values for comparisons are by Kruskal-wallis one-way analysis of variance by rank test.

Techniques Used: Staining, Expressing, Cell Culture

2) Product Images from "Human Immunodeficiency Virus-Specific CD8+ T Cells in Human Breast Milk"

Article Title: Human Immunodeficiency Virus-Specific CD8+ T Cells in Human Breast Milk

Journal: Journal of Virology

doi: 10.1128/JVI.76.15.7365-7373.2002

HIV-1-specific immune responses to overlapping 20-mer peptide pools from Gag, Pol, Env, and Nef of HIV-1; tetanus toxoid (TT); or CMV lysate as measured by the IFN-γ ELISPOT assay. (A) BMC from HIV-seronegative women. (B) BMC responses from HIV + women (U.S. cohort). (C) BMC responses from HIV + women (Zambian cohort). Fifty peptides each, comprising the N terminus (Pol N aa 1 to 510) or the C terminus (Pol C aa 501 to 1003) of Pol were used instead of a pool of 100 peptides (Pol aa 1 to 1003) to stimulate BMC from the Zambian volunteers. Asterisks indicate that for volunteer 1 the responses from Pol N and Pol C were added and plotted as Pol, and the same was done for Env using gp120 and gp41. Responses to CMV and TT were not measured in volunteers 4 and 5. PHA responses were > 1,000 SFC/million BMC (data not shown).
Figure Legend Snippet: HIV-1-specific immune responses to overlapping 20-mer peptide pools from Gag, Pol, Env, and Nef of HIV-1; tetanus toxoid (TT); or CMV lysate as measured by the IFN-γ ELISPOT assay. (A) BMC from HIV-seronegative women. (B) BMC responses from HIV + women (U.S. cohort). (C) BMC responses from HIV + women (Zambian cohort). Fifty peptides each, comprising the N terminus (Pol N aa 1 to 510) or the C terminus (Pol C aa 501 to 1003) of Pol were used instead of a pool of 100 peptides (Pol aa 1 to 1003) to stimulate BMC from the Zambian volunteers. Asterisks indicate that for volunteer 1 the responses from Pol N and Pol C were added and plotted as Pol, and the same was done for Env using gp120 and gp41. Responses to CMV and TT were not measured in volunteers 4 and 5. PHA responses were > 1,000 SFC/million BMC (data not shown).

Techniques Used: Enzyme-linked Immunospot

3) Product Images from "Cytokine Profiles and Cell Proliferation Responses to Truncated ORF2 Protein in Iranian Patients Recovered from Hepatitis E Infection"

Article Title: Cytokine Profiles and Cell Proliferation Responses to Truncated ORF2 Protein in Iranian Patients Recovered from Hepatitis E Infection

Journal: Journal of Tropical Medicine

doi: 10.1155/2015/523560

The levels of IL-4, IL-10, IL-12 p70, and IFN- γ cytokines in HEV-recovered and control groups following stimulation with the truncated ORF2 protein. Significant amounts of IFN- γ and IL-12 cytokines but low amounts of IL-10 and IL-4 cytokines were produced. IL-12 p70 and IFN- γ cytokines production following stimulation with the truncated ORF2 protein was significantly higher in the recovered group compared to the control group, while there were no significant differences in the levels of IL-10 and IL-4 between the two groups. Both groups showed almost similar expression of all cytokines following stimulation with PHA. Results are shown as mean ± SD pg/mL.
Figure Legend Snippet: The levels of IL-4, IL-10, IL-12 p70, and IFN- γ cytokines in HEV-recovered and control groups following stimulation with the truncated ORF2 protein. Significant amounts of IFN- γ and IL-12 cytokines but low amounts of IL-10 and IL-4 cytokines were produced. IL-12 p70 and IFN- γ cytokines production following stimulation with the truncated ORF2 protein was significantly higher in the recovered group compared to the control group, while there were no significant differences in the levels of IL-10 and IL-4 between the two groups. Both groups showed almost similar expression of all cytokines following stimulation with PHA. Results are shown as mean ± SD pg/mL.

Techniques Used: Produced, Expressing

Cell proliferative responses in HEV-recovered individuals and control group. Proliferative responses to the truncated ORF2 protein are significantly higher in HEV-recovered group compared to the control group, while both groups showed good nonspecific stimulation with PHA. The results are shown as proliferation index (mean ± SD PI).
Figure Legend Snippet: Cell proliferative responses in HEV-recovered individuals and control group. Proliferative responses to the truncated ORF2 protein are significantly higher in HEV-recovered group compared to the control group, while both groups showed good nonspecific stimulation with PHA. The results are shown as proliferation index (mean ± SD PI).

Techniques Used:

IFN- γ ELISPOT responses in HEV-recovered and control groups. IFN- γ ELISPOT responses to the truncated ORF2 protein are significantly higher in the recovered group compared to the controls, while both groups showed good nonspecific stimulation with PHA. The results are shown as spot forming cells per 10 5 cells (SFC/10 5 cells).
Figure Legend Snippet: IFN- γ ELISPOT responses in HEV-recovered and control groups. IFN- γ ELISPOT responses to the truncated ORF2 protein are significantly higher in the recovered group compared to the controls, while both groups showed good nonspecific stimulation with PHA. The results are shown as spot forming cells per 10 5 cells (SFC/10 5 cells).

Techniques Used: Enzyme-linked Immunospot

4) Product Images from "In Vitro Exposure to Prostratin but Not Bryostatin-1 Improves Natural Killer Cell Functions Including Killing of CD4+ T Cells Harboring Reactivated Human Immunodeficiency Virus"

Article Title: In Vitro Exposure to Prostratin but Not Bryostatin-1 Improves Natural Killer Cell Functions Including Killing of CD4+ T Cells Harboring Reactivated Human Immunodeficiency Virus

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.01514

ULBP2 is induced on latently infected CD4 + T cells upon HIV-1 reactivation by prostratin (PRO) and, to a lesser extent, by bryostatin-1 (BRY). To establish viral latency, freshly isolated CD4 + T cells were cultivated in the presence of CCL19 for 1–3 days, infected or not with HIV-1, and further cultured for 3 days, as described in details in Section “ Materials and Methods .” Then, cells were stimulated with 10 µg/ml phytohemagglutinin (PHA), 1 µM PRO, or 10 nM BRY, or not stimulated (ns), further cultivated for 3 days and finally analyzed by two-color flow cytometry for the expression of intracellular p24 and cell-surface MICA/B and ULBP2. (A) Representative dot plots show the frequency of reactivated p24 + cells gated on non-infected PHA-stimulated control cells. (B) Percentage of p24 + cells was determined as shown in panel (A) in four independent experiments and normalized to HIV-infected PHA-treated cultures (mean ± SEM). (C) Histograms show MICA/B and ULBP2 fluorescence in the gated p24 − (top panels) and p24 + (bottom panels) cell populations measured in a representative experiment on ns, PRO- and BRY-stimulated cell samples. Signals obtained with control IgG (filled histograms) and the percentage of ligand-positive cells are shown. (D) ULBP2 expression (mean ± SEM), both% of positive cells and mean fluorescence intensity (MFI), was determined as shown in panel (C) in four independent experiments. (E) RT-qPCR was used to assess ULBP2 mRNA levels in latently infected and in control non-infected CD4 + T cells cultivated for 24 h in ns, PRO, and BRY conditions. Results are expressed relative to ns non-infected sample (set to 1). Mean ± SEM values obtained in three independent experiments performed in duplicate are shown. * P
Figure Legend Snippet: ULBP2 is induced on latently infected CD4 + T cells upon HIV-1 reactivation by prostratin (PRO) and, to a lesser extent, by bryostatin-1 (BRY). To establish viral latency, freshly isolated CD4 + T cells were cultivated in the presence of CCL19 for 1–3 days, infected or not with HIV-1, and further cultured for 3 days, as described in details in Section “ Materials and Methods .” Then, cells were stimulated with 10 µg/ml phytohemagglutinin (PHA), 1 µM PRO, or 10 nM BRY, or not stimulated (ns), further cultivated for 3 days and finally analyzed by two-color flow cytometry for the expression of intracellular p24 and cell-surface MICA/B and ULBP2. (A) Representative dot plots show the frequency of reactivated p24 + cells gated on non-infected PHA-stimulated control cells. (B) Percentage of p24 + cells was determined as shown in panel (A) in four independent experiments and normalized to HIV-infected PHA-treated cultures (mean ± SEM). (C) Histograms show MICA/B and ULBP2 fluorescence in the gated p24 − (top panels) and p24 + (bottom panels) cell populations measured in a representative experiment on ns, PRO- and BRY-stimulated cell samples. Signals obtained with control IgG (filled histograms) and the percentage of ligand-positive cells are shown. (D) ULBP2 expression (mean ± SEM), both% of positive cells and mean fluorescence intensity (MFI), was determined as shown in panel (C) in four independent experiments. (E) RT-qPCR was used to assess ULBP2 mRNA levels in latently infected and in control non-infected CD4 + T cells cultivated for 24 h in ns, PRO, and BRY conditions. Results are expressed relative to ns non-infected sample (set to 1). Mean ± SEM values obtained in three independent experiments performed in duplicate are shown. * P

Techniques Used: Infection, Isolation, Cell Culture, Flow Cytometry, Cytometry, Expressing, Fluorescence, Quantitative RT-PCR

5) Product Images from "Impaired B cells survival upon production of inflammatory cytokines by HIV-1 exposed follicular dendritic cells"

Article Title: Impaired B cells survival upon production of inflammatory cytokines by HIV-1 exposed follicular dendritic cells

Journal: Retrovirology

doi: 10.1186/s12977-016-0295-4

Migration of B and T cells towards HIV-1 exposed FDCs. PBMCs from 3 donors were activated with CpG, PHA or the combination of anti IgM and anti CD40 and placed for 4 h on trans-well chambers containing the supernatant from FDCs (8–13 and 1403) exposed to HIV-1 (IIIB or SF162 strain) for 7 days. Cells were also migrated towards medium. The frequency of migrated CD19+ B ( a – c ) and CD3+ T-cells ( d – f ) in total migrated cells were measured by Flow cytometry. The data represent the cumulative mean and standard deviation for 3 activation conditions in cells from 3 donors
Figure Legend Snippet: Migration of B and T cells towards HIV-1 exposed FDCs. PBMCs from 3 donors were activated with CpG, PHA or the combination of anti IgM and anti CD40 and placed for 4 h on trans-well chambers containing the supernatant from FDCs (8–13 and 1403) exposed to HIV-1 (IIIB or SF162 strain) for 7 days. Cells were also migrated towards medium. The frequency of migrated CD19+ B ( a – c ) and CD3+ T-cells ( d – f ) in total migrated cells were measured by Flow cytometry. The data represent the cumulative mean and standard deviation for 3 activation conditions in cells from 3 donors

Techniques Used: Migration, Flow Cytometry, Cytometry, Standard Deviation, Activation Assay

6) Product Images from "Synergistic Activation of Latent HIV-1 Expression by Novel Histone Deacetylase Inhibitors and Bryostatin-1"

Article Title: Synergistic Activation of Latent HIV-1 Expression by Novel Histone Deacetylase Inhibitors and Bryostatin-1

Journal: Scientific Reports

doi: 10.1038/srep16445

Primary human cells phenotype after treatment with selected drug combinations. Purified CD4 T cells from healthy subjects were treated with the indicated concentrations of BRY, PNB, RMD or with PHA or PMA/ionomycin for 1 (grey bars) and 3 (black bars) days. ( a ) Cell viability was determined using 7AAD reagent and analysed by flow cytometry and expressed as percentage. ( b ) The surface expression of the activation markers CD38 and CD69 in viable CD4 T cells were analysed by flow cytometry and expressed as iMFI. ( c ) Cell proliferation was determined as BrdU incorporation measured by ELISA. Results are normalized to control vehicle-treated CD4 T cells from the same donors and represent the mean + SEM of three independent experiments.
Figure Legend Snippet: Primary human cells phenotype after treatment with selected drug combinations. Purified CD4 T cells from healthy subjects were treated with the indicated concentrations of BRY, PNB, RMD or with PHA or PMA/ionomycin for 1 (grey bars) and 3 (black bars) days. ( a ) Cell viability was determined using 7AAD reagent and analysed by flow cytometry and expressed as percentage. ( b ) The surface expression of the activation markers CD38 and CD69 in viable CD4 T cells were analysed by flow cytometry and expressed as iMFI. ( c ) Cell proliferation was determined as BrdU incorporation measured by ELISA. Results are normalized to control vehicle-treated CD4 T cells from the same donors and represent the mean + SEM of three independent experiments.

Techniques Used: Purification, Flow Cytometry, Cytometry, Expressing, Activation Assay, BrdU Incorporation Assay, Enzyme-linked Immunosorbent Assay

7) Product Images from "Anti-HIV Activity of Human Defensin 5 in Primary CD4+ T Cells under Serum-Deprived Conditions Is a Consequence of Defensin-Mediated Cytotoxicity"

Article Title: Anti-HIV Activity of Human Defensin 5 in Primary CD4+ T Cells under Serum-Deprived Conditions Is a Consequence of Defensin-Mediated Cytotoxicity

Journal: PLoS ONE

doi: 10.1371/journal.pone.0076038

Serum deprivation contributes to anti-HIV activity of HD5 in primary CD4+ T cells. HIV-1 primary isolate 20635–4 (R5 virus, clade C) was incubated with HD5 at different concentrations for 1 h at 37°C. CD4+ T cells from PHA-activated PBLs (A) and PHA-activated CD4+ T cells (B) were incubated with virus-defensin mixture for 2 h at 37°C (no spinoculation) or 1.5 h at 1250×g spinning (spinoculation). After washing off unbound virus, cells were cultured in RPMI containing 10% FBS and IL-2 or 0.3% human AB serum, ITS supplement, and IL-2. HD5 was added back to cell cultures in the presence of IL-2. The level of p24 protein in culture media was measured by p24 ELISA. Data presented are the average ± standard deviation of 3 replicates. Similar results were observed in 3 independent experiments from different donors; *P
Figure Legend Snippet: Serum deprivation contributes to anti-HIV activity of HD5 in primary CD4+ T cells. HIV-1 primary isolate 20635–4 (R5 virus, clade C) was incubated with HD5 at different concentrations for 1 h at 37°C. CD4+ T cells from PHA-activated PBLs (A) and PHA-activated CD4+ T cells (B) were incubated with virus-defensin mixture for 2 h at 37°C (no spinoculation) or 1.5 h at 1250×g spinning (spinoculation). After washing off unbound virus, cells were cultured in RPMI containing 10% FBS and IL-2 or 0.3% human AB serum, ITS supplement, and IL-2. HD5 was added back to cell cultures in the presence of IL-2. The level of p24 protein in culture media was measured by p24 ELISA. Data presented are the average ± standard deviation of 3 replicates. Similar results were observed in 3 independent experiments from different donors; *P

Techniques Used: Activity Assay, Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay, Standard Deviation

8) Product Images from "Shipping blood to a central laboratory in multicenter clinical trials: effect of ambient temperature on specimen temperature, and effects of temperature on mononuclear cell yield, viability and immunologic function"

Article Title: Shipping blood to a central laboratory in multicenter clinical trials: effect of ambient temperature on specimen temperature, and effects of temperature on mononuclear cell yield, viability and immunologic function

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-9-26

Mitogen and antigen-activated PBMC responses as detected by IFNgamma secretion in an ELIspot assay . After thawing from liquid nitrogen, PBMC were incubated 18 hours at 37°C with (A) PMA/ionomycin, (B) PHA or (C) CEF peptide pool and then tested for IFNg secretion by ELIspot assay. Results are presented as SFC per 200,000 PBMC for PMA and PHA. CEF SFC are adjusted for the percentage of CD8+ T cells and presented as SFC per 200,000 CD8 T cells. Each condition is compared to the control condition (arrows) as described in Figure 3. (*) p
Figure Legend Snippet: Mitogen and antigen-activated PBMC responses as detected by IFNgamma secretion in an ELIspot assay . After thawing from liquid nitrogen, PBMC were incubated 18 hours at 37°C with (A) PMA/ionomycin, (B) PHA or (C) CEF peptide pool and then tested for IFNg secretion by ELIspot assay. Results are presented as SFC per 200,000 PBMC for PMA and PHA. CEF SFC are adjusted for the percentage of CD8+ T cells and presented as SFC per 200,000 CD8 T cells. Each condition is compared to the control condition (arrows) as described in Figure 3. (*) p

Techniques Used: Enzyme-linked Immunospot, Incubation

9) Product Images from "hCTLA4-Gene-Modified Human Bone Marrow-Derived Mesenchymal Stem Cells (hBMMSCs) Maintain POSTN Secretion to Enhance the Migration Capability of Allogeneic hBMMSCs through the Integrin αvβ3/FAK/ERK Signaling Pathway"

Article Title: hCTLA4-Gene-Modified Human Bone Marrow-Derived Mesenchymal Stem Cells (hBMMSCs) Maintain POSTN Secretion to Enhance the Migration Capability of Allogeneic hBMMSCs through the Integrin αvβ3/FAK/ERK Signaling Pathway

Journal: Stem Cells International

doi: 10.1155/2020/3608284

POSTN levels in the coculture supernatant. NC hBMMSCs, CTLA4-modified hBMMSCs, POSTN-modified hBMMSCs, or CTLA4+shPOSTN-modified hBMMSCs were cocultured with PBMCs treated with PHA for 72 h, and the coculture supernatant was isolated to detect the POSTN level by ELISA. ELISA was performed using the coculture supernatant isolated from three independent cell treatments. ∗ P
Figure Legend Snippet: POSTN levels in the coculture supernatant. NC hBMMSCs, CTLA4-modified hBMMSCs, POSTN-modified hBMMSCs, or CTLA4+shPOSTN-modified hBMMSCs were cocultured with PBMCs treated with PHA for 72 h, and the coculture supernatant was isolated to detect the POSTN level by ELISA. ELISA was performed using the coculture supernatant isolated from three independent cell treatments. ∗ P

Techniques Used: Modification, Isolation, Enzyme-linked Immunosorbent Assay

Migration capability of allo-hBMMSCs was enhanced by the culture supernatant of CTLA4- and POSTN-modified hBMMSCs cocultured with PBMCs treated with PHA. The concentrated culture supernatant from NC hBMMSCs, CTLA4-modified hBMMSCs, POSTN-modified hBMMSCs, or CTLA4+shPOSTN-modified hBMMSCs cocultured with PBMCs treated with PHA was added into the culture medium of allo-hBMMSCs in a ratio of 1 : 10, and the treated allo-hBMMSCs were named as NC, CTLA4, POSTN, and CTLA4+shPOSTN, according to their treatment. The migration capability of allo-hBMMSCs in each group was evaluated by Transwell assay (a), wound healing assay (b), and western blot to detect MMP2 and MMP9 levels (c). For all panels, left is the representative graphs and right is the statistical results based on three independent experiments. ∗ P
Figure Legend Snippet: Migration capability of allo-hBMMSCs was enhanced by the culture supernatant of CTLA4- and POSTN-modified hBMMSCs cocultured with PBMCs treated with PHA. The concentrated culture supernatant from NC hBMMSCs, CTLA4-modified hBMMSCs, POSTN-modified hBMMSCs, or CTLA4+shPOSTN-modified hBMMSCs cocultured with PBMCs treated with PHA was added into the culture medium of allo-hBMMSCs in a ratio of 1 : 10, and the treated allo-hBMMSCs were named as NC, CTLA4, POSTN, and CTLA4+shPOSTN, according to their treatment. The migration capability of allo-hBMMSCs in each group was evaluated by Transwell assay (a), wound healing assay (b), and western blot to detect MMP2 and MMP9 levels (c). For all panels, left is the representative graphs and right is the statistical results based on three independent experiments. ∗ P

Techniques Used: Migration, Modification, Transwell Assay, Wound Healing Assay, Western Blot

Integrin α v β 3/FAK/ERK signaling pathway was blocked by anti-integrin α v β 3 treatment. Approximately 2 μ g/mL negative control IgG or anti-integrin α v β 3 was added into the culture medium of allo-hBMMSCs with the concentrated culture supernatant from CTLA4-modified hBMMSCs or POSTN-modified hBMMSCs and allo-hBMMSCs cocultured with PBMCs treated with PHA, and treated allo-hBMMSCs were named CTLA4+IgG, CTLA4+anti-integrin α v β 3, POSTN+IgG, or POSTN+anti-integrin α v β 3. Protein levels of key factors in the integrin α v β 3/FAK/ERK signaling pathway were detected by western blotting. (a) Representative graphs of western blotting and (b) statistical results of relative protein expression based on three independent experiments. ∗ P
Figure Legend Snippet: Integrin α v β 3/FAK/ERK signaling pathway was blocked by anti-integrin α v β 3 treatment. Approximately 2 μ g/mL negative control IgG or anti-integrin α v β 3 was added into the culture medium of allo-hBMMSCs with the concentrated culture supernatant from CTLA4-modified hBMMSCs or POSTN-modified hBMMSCs and allo-hBMMSCs cocultured with PBMCs treated with PHA, and treated allo-hBMMSCs were named CTLA4+IgG, CTLA4+anti-integrin α v β 3, POSTN+IgG, or POSTN+anti-integrin α v β 3. Protein levels of key factors in the integrin α v β 3/FAK/ERK signaling pathway were detected by western blotting. (a) Representative graphs of western blotting and (b) statistical results of relative protein expression based on three independent experiments. ∗ P

Techniques Used: Negative Control, Modification, Western Blot, Expressing

Migration capability of allo-hBMMSCs in each group was suppressed by anti-integrin α v β 3 treatment. Approximately 2 μ g/mL negative control IgG or anti-integrin α v β 3 was added to the culture medium of allo-hBMMSCs with concentrated culture supernatant from CTLA4-modified hBMMSCs or POSTN-modified hBMMSCs and allo-hBMMSCs co-cultured with PBMCs treated with PHA, and the treated allo-hBMMSCs were named CTLA4+IgG, CTLA4+anti-integrin α v β 3, POSTN+IgG, or POSTN+anti-integrin α v β 3. The migration capability of allo-hBMMSCs in each group was evaluated by Transwell assay (a), wound healing assay (b), and western blotting to detect MMP2 and MMP9 levels (c). For all panels, left shows the representative images or blots and right shows the statistical results based on three independent experiments. ∗ P
Figure Legend Snippet: Migration capability of allo-hBMMSCs in each group was suppressed by anti-integrin α v β 3 treatment. Approximately 2 μ g/mL negative control IgG or anti-integrin α v β 3 was added to the culture medium of allo-hBMMSCs with concentrated culture supernatant from CTLA4-modified hBMMSCs or POSTN-modified hBMMSCs and allo-hBMMSCs co-cultured with PBMCs treated with PHA, and the treated allo-hBMMSCs were named CTLA4+IgG, CTLA4+anti-integrin α v β 3, POSTN+IgG, or POSTN+anti-integrin α v β 3. The migration capability of allo-hBMMSCs in each group was evaluated by Transwell assay (a), wound healing assay (b), and western blotting to detect MMP2 and MMP9 levels (c). For all panels, left shows the representative images or blots and right shows the statistical results based on three independent experiments. ∗ P

Techniques Used: Migration, Negative Control, Modification, Cell Culture, Transwell Assay, Wound Healing Assay, Western Blot

Levels of IL-2 and IFN- γ in the culture supernatant of PBMCs treated with PHA. After treatment with or without PHA for 24, 48, 72, and 96 h, IL-2 and IFN- γ levels were detected by ELISA. ELISA was performed using the culture supernatant isolated from three independent cell treatments. ∗ P
Figure Legend Snippet: Levels of IL-2 and IFN- γ in the culture supernatant of PBMCs treated with PHA. After treatment with or without PHA for 24, 48, 72, and 96 h, IL-2 and IFN- γ levels were detected by ELISA. ELISA was performed using the culture supernatant isolated from three independent cell treatments. ∗ P

Techniques Used: Enzyme-linked Immunosorbent Assay, Isolation

The integrin α v β 3/FAK/ERK signaling pathway in allo-hBMMSCs was activated by the culture supernatant of CTLA4- and POSTN-modified hBMMSCs cocultured with PBMCs treated with PHA. The concentrated culture supernatant from NC hBMMSCs, CTLA4-modified hBMMSCs, POSTN-modified hBMMSCs, or CTLA4+shPOSTN-modified hBMMSCs cocultured with PBMCs treated with PHA was added into the culture medium of allo-hBMMSCs at a ratio of 1 : 10, and the treated allo-hBMMSCs were named NC, CTLA4, POSTN, and CTLA4+shPOSTN, according to their treatment. Protein levels of key factors in the integrin α v β 3/FAK/ERK signaling pathway were detected by western blotting. (a) Representative graphs of western blotting and (b) statistical results of relative protein expression based on three independent experiments. ∗ P
Figure Legend Snippet: The integrin α v β 3/FAK/ERK signaling pathway in allo-hBMMSCs was activated by the culture supernatant of CTLA4- and POSTN-modified hBMMSCs cocultured with PBMCs treated with PHA. The concentrated culture supernatant from NC hBMMSCs, CTLA4-modified hBMMSCs, POSTN-modified hBMMSCs, or CTLA4+shPOSTN-modified hBMMSCs cocultured with PBMCs treated with PHA was added into the culture medium of allo-hBMMSCs at a ratio of 1 : 10, and the treated allo-hBMMSCs were named NC, CTLA4, POSTN, and CTLA4+shPOSTN, according to their treatment. Protein levels of key factors in the integrin α v β 3/FAK/ERK signaling pathway were detected by western blotting. (a) Representative graphs of western blotting and (b) statistical results of relative protein expression based on three independent experiments. ∗ P

Techniques Used: Modification, Western Blot, Expressing

10) Product Images from "Inhibition of mesenchymal stromal cells by pre-activated lymphocytes and their culture media"

Article Title: Inhibition of mesenchymal stromal cells by pre-activated lymphocytes and their culture media

Journal: Stem Cell Research & Therapy

doi: 10.1186/scrt392

Impedance profiles of MSCs in co-culture with NK-depleted PBMCs. Unstimulated (NS) NK-depleted PBMCs (A) and PHA-stimulated NK-depleted PBMCs (B) were added to MSCs (400/well), after about 72 hours of culture, at four different ratios. The experiment was conducted in triplicate. MSCs, mesenchymal stem cells; NK, natural killer; PBMCs, peripheral blood mononuclear cells; PHA, phytohemagglutinin.
Figure Legend Snippet: Impedance profiles of MSCs in co-culture with NK-depleted PBMCs. Unstimulated (NS) NK-depleted PBMCs (A) and PHA-stimulated NK-depleted PBMCs (B) were added to MSCs (400/well), after about 72 hours of culture, at four different ratios. The experiment was conducted in triplicate. MSCs, mesenchymal stem cells; NK, natural killer; PBMCs, peripheral blood mononuclear cells; PHA, phytohemagglutinin.

Techniques Used: Co-Culture Assay

Impedance profiles and mean CI values of MSCs treated with lymphocyte conditioned medium (CM). MSCs were stimulated with CM from unstimulated lymphocytes (A) and PHA-stimulated lymphocytes (B) . The CM was added to MSCs (400/well) at four different dilutions after about 72 hours of culture. The experiment was conducted in triplicate. (C) The mean CI values, obtained in the four independent experiments, treating MSCs with CM from resting or pre-activated PBMCs. Results were analyzed and expressed as percentage of the untreated MSCs and presented as mean values ± standard deviations. Statistical analysis was performed using one-way ANOVA (*P
Figure Legend Snippet: Impedance profiles and mean CI values of MSCs treated with lymphocyte conditioned medium (CM). MSCs were stimulated with CM from unstimulated lymphocytes (A) and PHA-stimulated lymphocytes (B) . The CM was added to MSCs (400/well) at four different dilutions after about 72 hours of culture. The experiment was conducted in triplicate. (C) The mean CI values, obtained in the four independent experiments, treating MSCs with CM from resting or pre-activated PBMCs. Results were analyzed and expressed as percentage of the untreated MSCs and presented as mean values ± standard deviations. Statistical analysis was performed using one-way ANOVA (*P

Techniques Used:

MTT cell viability assay. Cell viability was assessed after MSC treatment with the indicated dose of lymphocyte CM. (A) CM from unstimulated PBMCs; (B) CM from PHA-stimulated PBMCs. Results were analyzed and expressed as percentage of the control (untreated MSCs) and presented as mean values ± standard deviations of the three independent experiments performed in duplicate. Statistical analysis was performed using Student’s t test (* P
Figure Legend Snippet: MTT cell viability assay. Cell viability was assessed after MSC treatment with the indicated dose of lymphocyte CM. (A) CM from unstimulated PBMCs; (B) CM from PHA-stimulated PBMCs. Results were analyzed and expressed as percentage of the control (untreated MSCs) and presented as mean values ± standard deviations of the three independent experiments performed in duplicate. Statistical analysis was performed using Student’s t test (* P

Techniques Used: MTT Assay, Viability Assay

Impedance profiles and mean CI values of MSCs in co-culture with PBMCs. Unstimulated (NS) PBMCs (A) and PHA-stimulated PBMCs (B) were added to MSCs (400/well) after about 72 hours of culture at four different ratios. The experiment was conducted in triplicate. (C) The mean CI values, obtained in the four independent experiments, treating MSCs with resting or pre-activated PBMCs. Results were analyzed and expressed as percentage of the untreated MSCs and presented as mean values ± standard deviations. Statistical analysis was performed using one-way ANOVA (* P
Figure Legend Snippet: Impedance profiles and mean CI values of MSCs in co-culture with PBMCs. Unstimulated (NS) PBMCs (A) and PHA-stimulated PBMCs (B) were added to MSCs (400/well) after about 72 hours of culture at four different ratios. The experiment was conducted in triplicate. (C) The mean CI values, obtained in the four independent experiments, treating MSCs with resting or pre-activated PBMCs. Results were analyzed and expressed as percentage of the untreated MSCs and presented as mean values ± standard deviations. Statistical analysis was performed using one-way ANOVA (* P

Techniques Used: Co-Culture Assay

11) Product Images from "Maternal Trypanosoma cruzi Infection Upregulates Capacity of Uninfected Neonate Cells To Produce Pro- and Anti-Inflammatory Cytokines"

Article Title: Maternal Trypanosoma cruzi Infection Upregulates Capacity of Uninfected Neonate Cells To Produce Pro- and Anti-Inflammatory Cytokines

Journal: Infection and Immunity

doi:

Production of anti-inflammatory cytokine and factors by WBC from T. cruzi -infected (■) or uninfected (□) mothers and their newborns. WBC were stimulated or not for 24 or 72 h with LPS (10 ng/ml) plus PHA (5 μg/ml) (A, C, and E) or a lysate of T. cruzi (10 6 lysed parasites/ml) (B, D, and F). Numbers of individuals in each group ranged from 22 to 41 for uninfected mothers, 9 to 15 for infected mothers, 26 to 43 for neonates from uninfected mothers, and 8 to 13 for uninfected neonates from infected mothers. Results (mean ± SEM) are expressed as the difference between levels obtained in stimulated and unstimulated cells. The Mann-Whitney-Wilcoxon U test was used for statistical comparisons between infected and uninfected mothers and between their newborns (*, P
Figure Legend Snippet: Production of anti-inflammatory cytokine and factors by WBC from T. cruzi -infected (■) or uninfected (□) mothers and their newborns. WBC were stimulated or not for 24 or 72 h with LPS (10 ng/ml) plus PHA (5 μg/ml) (A, C, and E) or a lysate of T. cruzi (10 6 lysed parasites/ml) (B, D, and F). Numbers of individuals in each group ranged from 22 to 41 for uninfected mothers, 9 to 15 for infected mothers, 26 to 43 for neonates from uninfected mothers, and 8 to 13 for uninfected neonates from infected mothers. Results (mean ± SEM) are expressed as the difference between levels obtained in stimulated and unstimulated cells. The Mann-Whitney-Wilcoxon U test was used for statistical comparisons between infected and uninfected mothers and between their newborns (*, P

Techniques Used: Infection, MANN-WHITNEY

Production of proinflammatory cytokines by WBC from T. cruzi -infected (■) or uninfected (□) mothers and their newborns. WBC were stimulated or not for 24 or 72 h with LPS (10 ng/ml) plus PHA (5 μg/ml) (A, C, and E) or a lysate of T. cruzi (10 6 lysed parasites/ml) (B, D, and F). Numbers of individuals in each group ranged from 23 to 42 for uninfected mothers, 7 to 13 for infected mothers, 16 to 46 for neonates from uninfected mothers, and 8 to 15 for uninfected neonates from infected mothers. Results (mean ± SEM) are expressed as the difference between levels obtained for stimulated and unstimulated cells. The Mann-Whitney-Wilcoxon U test was used for statistical comparisons between infected and uninfected mothers and between their newborns (∗, P
Figure Legend Snippet: Production of proinflammatory cytokines by WBC from T. cruzi -infected (■) or uninfected (□) mothers and their newborns. WBC were stimulated or not for 24 or 72 h with LPS (10 ng/ml) plus PHA (5 μg/ml) (A, C, and E) or a lysate of T. cruzi (10 6 lysed parasites/ml) (B, D, and F). Numbers of individuals in each group ranged from 23 to 42 for uninfected mothers, 7 to 13 for infected mothers, 16 to 46 for neonates from uninfected mothers, and 8 to 15 for uninfected neonates from infected mothers. Results (mean ± SEM) are expressed as the difference between levels obtained for stimulated and unstimulated cells. The Mann-Whitney-Wilcoxon U test was used for statistical comparisons between infected and uninfected mothers and between their newborns (∗, P

Techniques Used: Infection, MANN-WHITNEY

Production of type 1 and type 2 cytokines by WBC from T. cruzi -infected (■) or uninfected (□) mothers and their newborns. WBC were stimulated or not for 24 or 72 h with LPS (10 ng/ml) plus PHA (5 μg/ml) (A, C, E, and G) or a lysate of T. cruzi (10 6 lysed parasites/ml) (B, D, F, and H). Numbers of individuals in each group ranged from 26 to 43 for uninfected mothers, 11 to 16 for infected mothers, 25 to 47 for neonates from uninfected mothers, and 7 to 13 for uninfected neonates from infected mothers. Results (mean ± SEM) are expressed as the differences between levels obtained for stimulated and unstimulated cells. The Mann-Whitney-Wilcoxon U test was used for statistical comparisons between infected and uninfected mothers and between their newborns (∗, P
Figure Legend Snippet: Production of type 1 and type 2 cytokines by WBC from T. cruzi -infected (■) or uninfected (□) mothers and their newborns. WBC were stimulated or not for 24 or 72 h with LPS (10 ng/ml) plus PHA (5 μg/ml) (A, C, E, and G) or a lysate of T. cruzi (10 6 lysed parasites/ml) (B, D, F, and H). Numbers of individuals in each group ranged from 26 to 43 for uninfected mothers, 11 to 16 for infected mothers, 25 to 47 for neonates from uninfected mothers, and 7 to 13 for uninfected neonates from infected mothers. Results (mean ± SEM) are expressed as the differences between levels obtained for stimulated and unstimulated cells. The Mann-Whitney-Wilcoxon U test was used for statistical comparisons between infected and uninfected mothers and between their newborns (∗, P

Techniques Used: Infection, MANN-WHITNEY

12) Product Images from "Gnidimacrin, the Potent Anti-HIV Diterpene Can Eliminate Latent HIV-1 Ex Vivo by Activation of PKC Beta"

Article Title: Gnidimacrin, the Potent Anti-HIV Diterpene Can Eliminate Latent HIV-1 Ex Vivo by Activation of PKC Beta

Journal: Journal of medicinal chemistry

doi: 10.1021/acs.jmedchem.5b01233

GM activated latent HIV-1 in PBMCs. PBMCs from Pt-1 and Pt-6 were treated with GM (1 nM), SAHA (0.5 uM), or PHA (0.5 ug/mL). The PCR products from supernatants were analyzed with 2% agarose gel electrophoresis and documented with a Kodak molecular imager.
Figure Legend Snippet: GM activated latent HIV-1 in PBMCs. PBMCs from Pt-1 and Pt-6 were treated with GM (1 nM), SAHA (0.5 uM), or PHA (0.5 ug/mL). The PCR products from supernatants were analyzed with 2% agarose gel electrophoresis and documented with a Kodak molecular imager.

Techniques Used: Polymerase Chain Reaction, Agarose Gel Electrophoresis

13) Product Images from "Modulation of Human Immunodeficiency Virus (HIV)-Specific Immune Response by Using Efavirenz, Nelfinavir, and Stavudine in a Rescue Therapy Regimen for HIV-Infected, Drug-Experienced Patients"

Article Title: Modulation of Human Immunodeficiency Virus (HIV)-Specific Immune Response by Using Efavirenz, Nelfinavir, and Stavudine in a Rescue Therapy Regimen for HIV-Infected, Drug-Experienced Patients

Journal: Clinical and Diagnostic Laboratory Immunology

doi: 10.1128/CDLI.9.5.1114-1118.2002

(A) Env-stimulated cytokine production (IL-2, IFN-γ, IL-4, and IL-10) by PBMC of HIV-infected individuals at different time points (before treatment [t0] and 2 weeks [t1], 2 months [t2], and 10 months [t3] into therapy). The results are shown as means ± standard errors. (B) PHA-stimulated production of the same cytokines. *, values are significantly different from those at t0.
Figure Legend Snippet: (A) Env-stimulated cytokine production (IL-2, IFN-γ, IL-4, and IL-10) by PBMC of HIV-infected individuals at different time points (before treatment [t0] and 2 weeks [t1], 2 months [t2], and 10 months [t3] into therapy). The results are shown as means ± standard errors. (B) PHA-stimulated production of the same cytokines. *, values are significantly different from those at t0.

Techniques Used: Infection

14) Product Images from "Immuno-Modulatory and Anti-Inflammatory Effects of Dihydrogracilin A, a Terpene Derived from the Marine Sponge Dendrilla membranosa"

Article Title: Immuno-Modulatory and Anti-Inflammatory Effects of Dihydrogracilin A, a Terpene Derived from the Marine Sponge Dendrilla membranosa

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms18081643

Cytokine secretion profile of DHG-treated PBMC. PBMC of healthy donors ( n = 4) were activated with PHA (1.5%) for 24 h in the presence and in the absence of DHG at the indicated concentrations. Unstimulated cells are included as control (PBMC) in the figure. Supernatants were harvested and the concentrations of interleukin 6 (IL-6) ( A ), tumor necrosis factor-alpha (TNF-α) ( B ) and IL-10 ( C ) determined by ELISA immunoassay. Values reported refer to mean ± SD of four different donors. Statistical analyses are reported (ANOVA; * p
Figure Legend Snippet: Cytokine secretion profile of DHG-treated PBMC. PBMC of healthy donors ( n = 4) were activated with PHA (1.5%) for 24 h in the presence and in the absence of DHG at the indicated concentrations. Unstimulated cells are included as control (PBMC) in the figure. Supernatants were harvested and the concentrations of interleukin 6 (IL-6) ( A ), tumor necrosis factor-alpha (TNF-α) ( B ) and IL-10 ( C ) determined by ELISA immunoassay. Values reported refer to mean ± SD of four different donors. Statistical analyses are reported (ANOVA; * p

Techniques Used: Enzyme-linked Immunosorbent Assay

9,11-Dihydrogracilin A (DHG) inhibits Peripheral Blood Mononuclear Cells (PBMC) proliferation and viability and induces apoptosis. ( A ) Unstimulated PBMC and phytohemagglutinin (PHA)-activated PBMC from healthy donors were treated with DHG at the indicated concentrations. Proliferation was measured after 18h of 3 H-thymidine incorporation (1 µCi). The counts per minutes (c.p.m.) ± the SD of the triplicates of five independent experiments are shown. (ANOVA * p
Figure Legend Snippet: 9,11-Dihydrogracilin A (DHG) inhibits Peripheral Blood Mononuclear Cells (PBMC) proliferation and viability and induces apoptosis. ( A ) Unstimulated PBMC and phytohemagglutinin (PHA)-activated PBMC from healthy donors were treated with DHG at the indicated concentrations. Proliferation was measured after 18h of 3 H-thymidine incorporation (1 µCi). The counts per minutes (c.p.m.) ± the SD of the triplicates of five independent experiments are shown. (ANOVA * p

Techniques Used:

Flow cytometric analysis of CD25 and CD69 surface expression on DHG-treated PBMC. PBMC from healthy donors ( n = 7) were stimulated with PHA (1.5%) in the presence and in the absence of DHG. Unstimulated cells are included as control (PBMC) in the figure. Following 24 h of activation, CD3+/CD56− ( black bars ) and CD3−/CD56+ ( gray bars ) populations were analyzed for CD25+ expression ( A ) and CD69+ expression ( B ) and compared by ANOVA (* p
Figure Legend Snippet: Flow cytometric analysis of CD25 and CD69 surface expression on DHG-treated PBMC. PBMC from healthy donors ( n = 7) were stimulated with PHA (1.5%) in the presence and in the absence of DHG. Unstimulated cells are included as control (PBMC) in the figure. Following 24 h of activation, CD3+/CD56− ( black bars ) and CD3−/CD56+ ( gray bars ) populations were analyzed for CD25+ expression ( A ) and CD69+ expression ( B ) and compared by ANOVA (* p

Techniques Used: Expressing, Activation Assay

15) Product Images from "Metallothionein suppresses collagen-induced arthritis via induction of TGF-? and down-regulation of proinflammatory mediators"

Article Title: Metallothionein suppresses collagen-induced arthritis via induction of TGF-? and down-regulation of proinflammatory mediators

Journal: Clinical and Experimental Immunology

doi: 10.1046/j.1365-2249.2002.01922.x

Metallothionein administration inhibited the proliferation of CII-specific lymphocytes upon in vitro restimulation with CII. Lymph node cells from mice sacrificed on day 34 postimmunization were stimulated in triplicate in the absence (□) or presence of 250 μg/ml CII (▪) or 5 μg/ml PHA (▪) for 72 h. The cells were cultured for another 16 h with 1 μCi/well 3 H-thymidine, harvested, and counted using a scintillation counter. The data are representative of three independent experiments with similar results (* P
Figure Legend Snippet: Metallothionein administration inhibited the proliferation of CII-specific lymphocytes upon in vitro restimulation with CII. Lymph node cells from mice sacrificed on day 34 postimmunization were stimulated in triplicate in the absence (□) or presence of 250 μg/ml CII (▪) or 5 μg/ml PHA (▪) for 72 h. The cells were cultured for another 16 h with 1 μCi/well 3 H-thymidine, harvested, and counted using a scintillation counter. The data are representative of three independent experiments with similar results (* P

Techniques Used: In Vitro, Mouse Assay, Cell Culture

16) Product Images from "G1 Cell Cycle Arrest and Extrinsic Apoptotic Mechanisms Underlying the Anti-Leukemic Activity of CDK7 Inhibitor BS-181"

Article Title: G1 Cell Cycle Arrest and Extrinsic Apoptotic Mechanisms Underlying the Anti-Leukemic Activity of CDK7 Inhibitor BS-181

Journal: Cancers

doi: 10.3390/cancers12123845

Effect of the combined BS-181 and rTRAIL treatment on normal peripheral T lymphocytes. ( a ) To measure the effect of BS-181 and rTRAIL in combination on IL-2-dependent proliferation of activated normal human T cells, human peripheral T cells after stimulation with 1.0 μg/mL PHA for 60 h were incubated at a cell density of 5 × 10 4 cells/well with indicated concentrations of BS-181 and rTRAIL in the presence of 25 U/mL of rIL-2 in a 96-well plate for 20 h and with MTT solution for an additional 4 h. Mean ± SD ( n = 3 with three replicates per independent experiment). ** p
Figure Legend Snippet: Effect of the combined BS-181 and rTRAIL treatment on normal peripheral T lymphocytes. ( a ) To measure the effect of BS-181 and rTRAIL in combination on IL-2-dependent proliferation of activated normal human T cells, human peripheral T cells after stimulation with 1.0 μg/mL PHA for 60 h were incubated at a cell density of 5 × 10 4 cells/well with indicated concentrations of BS-181 and rTRAIL in the presence of 25 U/mL of rIL-2 in a 96-well plate for 20 h and with MTT solution for an additional 4 h. Mean ± SD ( n = 3 with three replicates per independent experiment). ** p

Techniques Used: Incubation, MTT Assay

17) Product Images from "Reversible Human Immunodeficiency Virus Type-1 Latency in Primary Human Monocyte-Derived Macrophages Induced by Sustained M1 Polarization"

Article Title: Reversible Human Immunodeficiency Virus Type-1 Latency in Primary Human Monocyte-Derived Macrophages Induced by Sustained M1 Polarization

Journal: Scientific Reports

doi: 10.1038/s41598-018-32451-w

Selective induction of HIV-1 replication (or expression of VSV-G pseudotyped virus) in M1 2 MDM stimulated by allogeneic PHA blasts or their supernatant and its selective prevention by lamivudine/3TC. ( A ) Kinetics of RT activity. PHA blasts upregulated virus replication in M1 2 MDM, but not in CTR or M1-MDM; the arrowheads indicate the timing of PHA blast addition to the infected MDM cultures. Mean ± SEM of 7 independent donors is reported. Comparison of different time points between M1 vs . M1 2 plus PHA blasts was performed using One-way ANOVA. ***p
Figure Legend Snippet: Selective induction of HIV-1 replication (or expression of VSV-G pseudotyped virus) in M1 2 MDM stimulated by allogeneic PHA blasts or their supernatant and its selective prevention by lamivudine/3TC. ( A ) Kinetics of RT activity. PHA blasts upregulated virus replication in M1 2 MDM, but not in CTR or M1-MDM; the arrowheads indicate the timing of PHA blast addition to the infected MDM cultures. Mean ± SEM of 7 independent donors is reported. Comparison of different time points between M1 vs . M1 2 plus PHA blasts was performed using One-way ANOVA. ***p

Techniques Used: Expressing, Activity Assay, Infection

18) Product Images from "Characterization of immortalized human islet stromal cells reveals a MSC-like profile with pancreatic features"

Article Title: Characterization of immortalized human islet stromal cells reveals a MSC-like profile with pancreatic features

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-020-01649-z

hISCs inhibit PBMCs proliferation. a Phase-contrast microscopic view of activated PBMCs after 96 h of co-culture with hISCs (top) or BM-MSCs (bottom). PHA activation induces the proliferation of PBMCs, which form suspended cell clusters (on the left). The number of PBMCs clusters decreases inversely with the ratio MSC to PBMC. The proliferation of PBMCs cultured alone (CTRL) or with hISCs or BM-MSCs at four ratios (1:5, 1:10, 1:20, 1:40; MSC to PBMC) was quantified. PBMCs proliferation was reduced by hISCs ( b ) and BM-MSCs ( c ) in co-culture. The inhibitory effect of both cell types was higher with increasing ratio. Results are representative of four independent experiments. * p
Figure Legend Snippet: hISCs inhibit PBMCs proliferation. a Phase-contrast microscopic view of activated PBMCs after 96 h of co-culture with hISCs (top) or BM-MSCs (bottom). PHA activation induces the proliferation of PBMCs, which form suspended cell clusters (on the left). The number of PBMCs clusters decreases inversely with the ratio MSC to PBMC. The proliferation of PBMCs cultured alone (CTRL) or with hISCs or BM-MSCs at four ratios (1:5, 1:10, 1:20, 1:40; MSC to PBMC) was quantified. PBMCs proliferation was reduced by hISCs ( b ) and BM-MSCs ( c ) in co-culture. The inhibitory effect of both cell types was higher with increasing ratio. Results are representative of four independent experiments. * p

Techniques Used: Co-Culture Assay, Activation Assay, Cell Culture

19) Product Images from "FIP-fve Stimulates Cell Proliferation and Enhances IL-2 Release by Activating MAP2K3/p38α (MAPK14) Signaling Pathway in Jurkat E6-1 Cells"

Article Title: FIP-fve Stimulates Cell Proliferation and Enhances IL-2 Release by Activating MAP2K3/p38α (MAPK14) Signaling Pathway in Jurkat E6-1 Cells

Journal: Frontiers in Nutrition

doi: 10.3389/fnut.2022.881924

FIP-fve treatment results in Jurkat E6-1 cells Proliferation. Jurkat E6-1 cells were treated with FIP-fve (100 μg/ml) or PHA (5 μg/ml) for 12 h (A) and 24 h (B) . Cell viability was assessed by Cell Counting kit-8 assay (CCK-8). PHA was used as a positive control. Values are mean ± SD, n = 3, ** p
Figure Legend Snippet: FIP-fve treatment results in Jurkat E6-1 cells Proliferation. Jurkat E6-1 cells were treated with FIP-fve (100 μg/ml) or PHA (5 μg/ml) for 12 h (A) and 24 h (B) . Cell viability was assessed by Cell Counting kit-8 assay (CCK-8). PHA was used as a positive control. Values are mean ± SD, n = 3, ** p

Techniques Used: Cell Counting, CCK-8 Assay, Positive Control

FIP-fve induced IL-2 release in Jurkat E6-1 cells. Jurkat E6-1 cells were treated with or without FIP-fve (25, 50, 100, and 200 μg/ml) for 6 h. PHA (1.5 and 5 μg/ml) was used as a positive control. IL-2 release was detected by ELISA kit for human IL-2 carried out according to the operating manual of the kit (enzyme marker, optical density, 0). Values are mean ± SD, n = 3, * p
Figure Legend Snippet: FIP-fve induced IL-2 release in Jurkat E6-1 cells. Jurkat E6-1 cells were treated with or without FIP-fve (25, 50, 100, and 200 μg/ml) for 6 h. PHA (1.5 and 5 μg/ml) was used as a positive control. IL-2 release was detected by ELISA kit for human IL-2 carried out according to the operating manual of the kit (enzyme marker, optical density, 0). Values are mean ± SD, n = 3, * p

Techniques Used: Positive Control, Enzyme-linked Immunosorbent Assay, Marker

Gossypetin and losmapimod inhibit cell proliferation and IL-2 release in FIP-fve -induced Jurkat E6-1 cells. (A) Gossypetin and losmapimod attenuated proliferation of FIP-fve -induced Jurkat E6-1 cells. Cell viability was determined after treatment within RPMI 1640 medium supplemented with 10% fetal bovine serum FIP-fve (100 μg/ml) and/or Losmapimod (0.1 μM/ml) and/or Gossypetin (40 μM/ml) for 12 h. Values are mean ± standard deviation, n = 3. PHA (5 μg/ml) was used as a positive control. (B) Gossypetin and losmapimod inhibit FIP-fve -induced IL-2 release in Jurkat E6-1 cells. IL-2 release was detected by ELISA kit for human IL-2 after treatment with as in (A) for 6 h. Values are mean ± SD, n = 3, * p
Figure Legend Snippet: Gossypetin and losmapimod inhibit cell proliferation and IL-2 release in FIP-fve -induced Jurkat E6-1 cells. (A) Gossypetin and losmapimod attenuated proliferation of FIP-fve -induced Jurkat E6-1 cells. Cell viability was determined after treatment within RPMI 1640 medium supplemented with 10% fetal bovine serum FIP-fve (100 μg/ml) and/or Losmapimod (0.1 μM/ml) and/or Gossypetin (40 μM/ml) for 12 h. Values are mean ± standard deviation, n = 3. PHA (5 μg/ml) was used as a positive control. (B) Gossypetin and losmapimod inhibit FIP-fve -induced IL-2 release in Jurkat E6-1 cells. IL-2 release was detected by ELISA kit for human IL-2 after treatment with as in (A) for 6 h. Values are mean ± SD, n = 3, * p

Techniques Used: Standard Deviation, Positive Control, Enzyme-linked Immunosorbent Assay

20) Product Images from "Blockade of Store-Operated Calcium Entry Reduces IL-17/TNF Cytokine-Induced Inflammatory Response in Human Myoblasts"

Article Title: Blockade of Store-Operated Calcium Entry Reduces IL-17/TNF Cytokine-Induced Inflammatory Response in Human Myoblasts

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.03170

PBMC-myoblast interaction induces a strong production CCL20 and IL-6. PBMC and myoblasts were cultured alone or in co-culture at a ratio of 5 PBMCs for 1 myoblast for 48 h in the presence or not of PHA (5 μg/mL). CCL20 and IL-6 secretion by myoblasts was quantified by ELISA (A–F) . The contribution of direct cell-cell contact was investigated with a cell culture permeable insert (C,D) . PBMCs were pre-incubated for 24 h in presence or not of PHA and then exposed or not to an anti-IL-17 antibody and/or an anti-TNFα antibody for 3 h before being added to the myoblast cultures. Data are expressed as CCL20 and IL-6 supernatant level percentages compared to the non-activated pre-incubated PBMC—myoblast co-cultures (E,F) . Data are the mean of 6–14 independent experiments ± SEM; * p
Figure Legend Snippet: PBMC-myoblast interaction induces a strong production CCL20 and IL-6. PBMC and myoblasts were cultured alone or in co-culture at a ratio of 5 PBMCs for 1 myoblast for 48 h in the presence or not of PHA (5 μg/mL). CCL20 and IL-6 secretion by myoblasts was quantified by ELISA (A–F) . The contribution of direct cell-cell contact was investigated with a cell culture permeable insert (C,D) . PBMCs were pre-incubated for 24 h in presence or not of PHA and then exposed or not to an anti-IL-17 antibody and/or an anti-TNFα antibody for 3 h before being added to the myoblast cultures. Data are expressed as CCL20 and IL-6 supernatant level percentages compared to the non-activated pre-incubated PBMC—myoblast co-cultures (E,F) . Data are the mean of 6–14 independent experiments ± SEM; * p

Techniques Used: Cell Culture, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Incubation

21) Product Images from "Immunomudulatory effects of hydroalcoholic extract of Hypericum perforatum"

Article Title: Immunomudulatory effects of hydroalcoholic extract of Hypericum perforatum

Journal: Avicenna Journal of Phytomedicine

doi:

Effects of hydroalcoholic extract of Hypericum perforatum administration on lymphocytes proliferation and respiratory burst in phagocytic cells. Splenocytes were isolated from sensitized mice with SRBC . A) Splenocytes cultured with 50 μL PHA solution (1 mg/ml) for 72 h . Then, lymphocytes proliferation were evaluated by MTT test. B) Splenocytes with S. aureus suspension and NBT were mixed and incubated for 30 min as detailed under materials and methods. The reduced dye was extracted in dioxan and quantitated at 520 nm. The values were presented as mean ± SD. (* p
Figure Legend Snippet: Effects of hydroalcoholic extract of Hypericum perforatum administration on lymphocytes proliferation and respiratory burst in phagocytic cells. Splenocytes were isolated from sensitized mice with SRBC . A) Splenocytes cultured with 50 μL PHA solution (1 mg/ml) for 72 h . Then, lymphocytes proliferation were evaluated by MTT test. B) Splenocytes with S. aureus suspension and NBT were mixed and incubated for 30 min as detailed under materials and methods. The reduced dye was extracted in dioxan and quantitated at 520 nm. The values were presented as mean ± SD. (* p

Techniques Used: Isolation, Mouse Assay, Cell Culture, MTT Assay, Incubation

22) Product Images from "Simple nanoliposomes encapsulating Lycium barbarum polysaccharides as adjuvants improve humoral and cellular immunity in mice"

Article Title: Simple nanoliposomes encapsulating Lycium barbarum polysaccharides as adjuvants improve humoral and cellular immunity in mice

Journal: International Journal of Nanomedicine

doi: 10.2147/IJN.S136820

Proliferative responses of splenocytes responding to ( A ) antigen, ( B ) LPS, and ( C ) PHA ex vivo. Notes: BALB/c mice (n=4) were immunized three times as described in the “subcutaneous immunization protocol” section. Splenocytes were harvested 14 days and 28 days after the third immunization. Splenocyte proliferation was measured using MTT, and the SI was calculated. Data are expressed as mean ± SEM ( p
Figure Legend Snippet: Proliferative responses of splenocytes responding to ( A ) antigen, ( B ) LPS, and ( C ) PHA ex vivo. Notes: BALB/c mice (n=4) were immunized three times as described in the “subcutaneous immunization protocol” section. Splenocytes were harvested 14 days and 28 days after the third immunization. Splenocyte proliferation was measured using MTT, and the SI was calculated. Data are expressed as mean ± SEM ( p

Techniques Used: Ex Vivo, Mouse Assay, MTT Assay

23) Product Images from "The Tricyclodecan-9-yl-xanthogenate D609 Triggers Ceramide Increase and Enhances FasL-Induced Caspase-Dependent and -Independent Cell Death in T Lymphocytes"

Article Title: The Tricyclodecan-9-yl-xanthogenate D609 Triggers Ceramide Increase and Enhances FasL-Induced Caspase-Dependent and -Independent Cell Death in T Lymphocytes

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms13078834

D609 inhibits SMS and GCS activities and enhances FasL-induced cell death in PHA-activated human T lymphocytes. ( A , B ) Human PBL, derived from three healthy volunteers, were cultured for 6 days in the presence of PHA. Cells were next pre-incubated in the presence or absence of the indicated concentrations of D609 for one hour and further incubated for 2 h in the presence of 2.5 μM C6-NBD-ceramide. SMS ( A ) and GCS ( B ) activities were determined. ( C , D ) PHA-activated PBL were pre-incubated for 1 h with or without 40 μM zVAD-fmk and 50 μg/mL D609 as indicated. Cells were further incubated for 16 h in the presence or absence of 500 ng/mL FasL. ( C ) The percentages of hypodiploid cells were determined by flow cytometry. Basal hypodiploidy in untreated cells did not exceed 15% in all conditions and was subtracted from the values. Values are means ± S.E.M. of three independent experiments. ( D ) PS externalization was measured by flow cytometry after annexin-V/FITC and propidium iodide staining. Analysis was restricted to propidium iodide negative cells to exclude cellular debris derived from non-activated lymphocytes. Numbers indicate the percentage of cells labeled with annexin-V/FITC. Data are representative of three independent experiments.
Figure Legend Snippet: D609 inhibits SMS and GCS activities and enhances FasL-induced cell death in PHA-activated human T lymphocytes. ( A , B ) Human PBL, derived from three healthy volunteers, were cultured for 6 days in the presence of PHA. Cells were next pre-incubated in the presence or absence of the indicated concentrations of D609 for one hour and further incubated for 2 h in the presence of 2.5 μM C6-NBD-ceramide. SMS ( A ) and GCS ( B ) activities were determined. ( C , D ) PHA-activated PBL were pre-incubated for 1 h with or without 40 μM zVAD-fmk and 50 μg/mL D609 as indicated. Cells were further incubated for 16 h in the presence or absence of 500 ng/mL FasL. ( C ) The percentages of hypodiploid cells were determined by flow cytometry. Basal hypodiploidy in untreated cells did not exceed 15% in all conditions and was subtracted from the values. Values are means ± S.E.M. of three independent experiments. ( D ) PS externalization was measured by flow cytometry after annexin-V/FITC and propidium iodide staining. Analysis was restricted to propidium iodide negative cells to exclude cellular debris derived from non-activated lymphocytes. Numbers indicate the percentage of cells labeled with annexin-V/FITC. Data are representative of three independent experiments.

Techniques Used: Derivative Assay, Cell Culture, Incubation, Flow Cytometry, Cytometry, Staining, Labeling

24) Product Images from "Downregulation of PD-L1 via amide analogues of brefelamide: Alternatives to antibody-based cancer immunotherapy"

Article Title: Downregulation of PD-L1 via amide analogues of brefelamide: Alternatives to antibody-based cancer immunotherapy

Journal: Experimental and Therapeutic Medicine

doi: 10.3892/etm.2020.8553

Restoration of T cell function by TPFS-201. (A) PD-L1 expression in A549 cells treated with 100 U/ml interferon-γ (upper). PD-1 expression in Jurkat T cells stimulated with 12.5 ng/ml PMA and 0.25 µg/ml PHA (lower). (B) Decreased apoptosis and (C) enhanced IL-2-luciferase expression in Jurkat T cells co-cultured with TPFS-201-treated A549 cells. A549 cells were pretreated with interferon-γ alone or in combination with TPFS-201 and co-cultured with Jurkat T cells stimulated with 12.5 ng/ml PHA and 0.25 µg/ml PMA. Caspase 3/7 activities and IL-2-directed luciferase expression were determined as indicators of T cell function. Caspase 3/7 activities or luciferase expression in Jurkat T cells co-cultured with TPFS-201-treated A549 cells were normalized to that of T cells co-cultured with vehicle-treated A549 cells, which was arbitrarily set as 100%. Results are presented as the mean ± standard deviation from three independent experiments. * P
Figure Legend Snippet: Restoration of T cell function by TPFS-201. (A) PD-L1 expression in A549 cells treated with 100 U/ml interferon-γ (upper). PD-1 expression in Jurkat T cells stimulated with 12.5 ng/ml PMA and 0.25 µg/ml PHA (lower). (B) Decreased apoptosis and (C) enhanced IL-2-luciferase expression in Jurkat T cells co-cultured with TPFS-201-treated A549 cells. A549 cells were pretreated with interferon-γ alone or in combination with TPFS-201 and co-cultured with Jurkat T cells stimulated with 12.5 ng/ml PHA and 0.25 µg/ml PMA. Caspase 3/7 activities and IL-2-directed luciferase expression were determined as indicators of T cell function. Caspase 3/7 activities or luciferase expression in Jurkat T cells co-cultured with TPFS-201-treated A549 cells were normalized to that of T cells co-cultured with vehicle-treated A549 cells, which was arbitrarily set as 100%. Results are presented as the mean ± standard deviation from three independent experiments. * P

Techniques Used: Cell Function Assay, Expressing, Luciferase, Cell Culture, Standard Deviation

25) Product Images from "Targeting glycosylated PD-1 induces potent anti-tumor immunity"

Article Title: Targeting glycosylated PD-1 induces potent anti-tumor immunity

Journal: Cancer research

doi: 10.1158/0008-5472.CAN-19-3133

TCR activation induces alterations of specific glycoforms of PD-1. (A) LC-MS/MS-based analysis of N-glycopeptides containing N74-glycans. PD-1 was purified from Jurkat-PD-1 FLAG cells treated with or without PHA overnight. The cartoon symbols used for the glycans conform to the standard representation recommended by the Consortium for Functional Glycomics. (B) Immunoblot of PD-1 in Jurkat T cells treated with PHA for the indicated time. (C) Lectin immunoblot of PD-1 purified from Jurkat-PD-1 FLAG treated with or without PHA for 5 hours. (D) Quantification of mRNA levels of various glycosyltransferases by real-time PCR in Jurkat T cells stimulated with α−CD3/CD28 for the indicated time points. All data represent mean ± SD from at least three independent experiments. *, P
Figure Legend Snippet: TCR activation induces alterations of specific glycoforms of PD-1. (A) LC-MS/MS-based analysis of N-glycopeptides containing N74-glycans. PD-1 was purified from Jurkat-PD-1 FLAG cells treated with or without PHA overnight. The cartoon symbols used for the glycans conform to the standard representation recommended by the Consortium for Functional Glycomics. (B) Immunoblot of PD-1 in Jurkat T cells treated with PHA for the indicated time. (C) Lectin immunoblot of PD-1 purified from Jurkat-PD-1 FLAG treated with or without PHA for 5 hours. (D) Quantification of mRNA levels of various glycosyltransferases by real-time PCR in Jurkat T cells stimulated with α−CD3/CD28 for the indicated time points. All data represent mean ± SD from at least three independent experiments. *, P

Techniques Used: Activation Assay, Liquid Chromatography with Mass Spectroscopy, Purification, Functional Assay, Real-time Polymerase Chain Reaction

26) Product Images from "Depressed Interleukin-12 (IL-12), but not IL-18, Production in Response to a 30- or 32-Kilodalton Mycobacterial Antigen in Patients with Active Pulmonary Tuberculosis"

Article Title: Depressed Interleukin-12 (IL-12), but not IL-18, Production in Response to a 30- or 32-Kilodalton Mycobacterial Antigen in Patients with Active Pulmonary Tuberculosis

Journal: Infection and Immunity

doi:

Southern blot analysis of cytokine cDNA after in vitro stimulation with the 30- or 32-kDa Ag of M. tuberculosis . PBMC were isolated from healthy controls and TB patients and cultured for 0, 6 (for IL-12 p40 and IL-18), and 96 (for IFN-γ) h in the presence of the 30- or 32-kDa Ag (1 μg/ml). Following reverse transcription, a PCR with primers specific for each cytokine was performed. The PCR products were electrophoresed on a 2% agarose gel and hybridized with a digoxigenin-labeled probe. Lanes P, mRNA from PBMC stimulated with PHA or LPS; lanes O, mRNA from freshly isolated PBMC.
Figure Legend Snippet: Southern blot analysis of cytokine cDNA after in vitro stimulation with the 30- or 32-kDa Ag of M. tuberculosis . PBMC were isolated from healthy controls and TB patients and cultured for 0, 6 (for IL-12 p40 and IL-18), and 96 (for IFN-γ) h in the presence of the 30- or 32-kDa Ag (1 μg/ml). Following reverse transcription, a PCR with primers specific for each cytokine was performed. The PCR products were electrophoresed on a 2% agarose gel and hybridized with a digoxigenin-labeled probe. Lanes P, mRNA from PBMC stimulated with PHA or LPS; lanes O, mRNA from freshly isolated PBMC.

Techniques Used: Southern Blot, In Vitro, Isolation, Cell Culture, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Labeling

27) Product Images from "Dichloroacetate modulates cytokines toward T helper 1 function via induction of the interleukin-12-interferon-? pathway"

Article Title: Dichloroacetate modulates cytokines toward T helper 1 function via induction of the interleukin-12-interferon-? pathway

Journal: OncoTargets and therapy

doi: 10.2147/OTT.S56688

High concentrations of DCA reduced the viability and proliferation of splenocytes. Notes: Using MTT assay, high concentrations of DCA (50 mM and 100 mM) reduced the viability and proliferation of unstimulated or PHA-stimulated mouse splenocytes (* P
Figure Legend Snippet: High concentrations of DCA reduced the viability and proliferation of splenocytes. Notes: Using MTT assay, high concentrations of DCA (50 mM and 100 mM) reduced the viability and proliferation of unstimulated or PHA-stimulated mouse splenocytes (* P

Techniques Used: MTT Assay

28) Product Images from "Borrelia burgdorferi Stimulates the Production of Interleukin-10 in Peripheral Blood Mononuclear Cells from Uninfected Humans and Rhesus Monkeys"

Article Title: Borrelia burgdorferi Stimulates the Production of Interleukin-10 in Peripheral Blood Mononuclear Cells from Uninfected Humans and Rhesus Monkeys

Journal: Infection and Immunity

doi:

(A) IL-10 production induced by OspA in PBMC from healthy human donors. PBMC (3 × 10 6 /ml) from each of three donors were incubated with supplemented medium (RPMI), L-OspA or U-OspA (1 μg/ml), heat-killed JD1 (10 7 spirochetes/ml), LPS (1 μg/ml), or PHA (10 μg/ml) for 48 h. Each pair of bars (open and solid, light and dark hatched, and light and dark stippled) corresponds to one donor. Samples were incubated without (open, light-hatched, and light-stippled bars) or with (solid, dark-hatched, and dark-stippled bars) polymyxin B (50 μg/ml). IL-10 in the supernatant was determined by ELISA. Values are means of duplicate determinations. (B) IL-10 mRNA expression induced by OspA in PBMC from uninfected rhesus monkeys. PBMC (3 × 10 6 /ml) from uninfected rhesus monkeys were stimulated with L-OspA (1 μg/ml), U-OspA (1 μg/ml), or ConA (8 μg/ml) for 24 h. The induced mRNA levels of IL-10 were determined by RT-PCR. Responses are shown as fold increases over unstimulated PBMC. Each point represents the response of PBMC from an individual monkey, and the horizontal lines indicate geometric means. All values were normalized with respect to GAPDH mRNA levels.
Figure Legend Snippet: (A) IL-10 production induced by OspA in PBMC from healthy human donors. PBMC (3 × 10 6 /ml) from each of three donors were incubated with supplemented medium (RPMI), L-OspA or U-OspA (1 μg/ml), heat-killed JD1 (10 7 spirochetes/ml), LPS (1 μg/ml), or PHA (10 μg/ml) for 48 h. Each pair of bars (open and solid, light and dark hatched, and light and dark stippled) corresponds to one donor. Samples were incubated without (open, light-hatched, and light-stippled bars) or with (solid, dark-hatched, and dark-stippled bars) polymyxin B (50 μg/ml). IL-10 in the supernatant was determined by ELISA. Values are means of duplicate determinations. (B) IL-10 mRNA expression induced by OspA in PBMC from uninfected rhesus monkeys. PBMC (3 × 10 6 /ml) from uninfected rhesus monkeys were stimulated with L-OspA (1 μg/ml), U-OspA (1 μg/ml), or ConA (8 μg/ml) for 24 h. The induced mRNA levels of IL-10 were determined by RT-PCR. Responses are shown as fold increases over unstimulated PBMC. Each point represents the response of PBMC from an individual monkey, and the horizontal lines indicate geometric means. All values were normalized with respect to GAPDH mRNA levels.

Techniques Used: Incubation, Enzyme-linked Immunosorbent Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

29) Product Images from "Repression of the HIV-1 5? LTR promoter and inhibition of HIV-1 replication by using engineered zinc-finger transcription factors"

Article Title: Repression of the HIV-1 5? LTR promoter and inhibition of HIV-1 replication by using engineered zinc-finger transcription factors

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.252770699

Luciferase assays for HIV-1 5′ LTR activity in human T cells in the presence or absence of Tat, PMA/PHA, and control vector pcDNA3.1(−), HIVBA′-KOX, or HIVBA′.
Figure Legend Snippet: Luciferase assays for HIV-1 5′ LTR activity in human T cells in the presence or absence of Tat, PMA/PHA, and control vector pcDNA3.1(−), HIVBA′-KOX, or HIVBA′.

Techniques Used: Luciferase, Activity Assay, Plasmid Preparation

Luciferase assays for HIV-1 5′ LTR activity in human T cells in the presence or absence of Tat, PMA/PHA, and various ZFP repressor constructs (150 ng of plasmid). The control contained pcDNA3.1(−) in place of the ZFP expression plasmid.
Figure Legend Snippet: Luciferase assays for HIV-1 5′ LTR activity in human T cells in the presence or absence of Tat, PMA/PHA, and various ZFP repressor constructs (150 ng of plasmid). The control contained pcDNA3.1(−) in place of the ZFP expression plasmid.

Techniques Used: Luciferase, Activity Assay, Construct, Plasmid Preparation, Expressing

30) Product Images from "Longitudinal Tracking of Cytokines after Acute Exposure to Tuberculosis: Association of Distinct Cytokine Patterns with Protection and Disease Development ▿"

Article Title: Longitudinal Tracking of Cytokines after Acute Exposure to Tuberculosis: Association of Distinct Cytokine Patterns with Protection and Disease Development ▿

Journal:

doi: 10.1128/CVI.00289-07

Dynamic changes in the IFN-γ/IL-10 ratios for DHCs, HHCs, and their index TB cases over a 2-year follow-up. The dynamic changes in the IFN-γ/IL-10 ratio in response to CF protein (a) and PHA and LPS (c) are shown. The numbers in the DHC
Figure Legend Snippet: Dynamic changes in the IFN-γ/IL-10 ratios for DHCs, HHCs, and their index TB cases over a 2-year follow-up. The dynamic changes in the IFN-γ/IL-10 ratio in response to CF protein (a) and PHA and LPS (c) are shown. The numbers in the DHC

Techniques Used:

31) Product Images from "Antigen-Specific T-Cell Responses in Humans after Intranasal Immunization with a Meningococcal Serogroup B Outer Membrane Vesicle Vaccine"

Article Title: Antigen-Specific T-Cell Responses in Humans after Intranasal Immunization with a Meningococcal Serogroup B Outer Membrane Vesicle Vaccine

Journal: Infection and Immunity

doi:

Comparisons of T-cell proliferation (cpm) in response to M. bovis BCG as a control antigen (20 or 4 μg/ml) and PHA (5 μg/ml) in 12 volunteers before and after intranasal immunizations with a meningococcal OMV vaccine without adjuvant. Each symbol represents the preimmunization cpm ( x value) and the maximum postimmunization cpm ( y value) obtained for one vaccinee. The diagonal solid line in each graph indicates the same post- as preimmunization values, and the two broken lines indicate twofold increases and decreases. The dotted horizontal lines define postimmunization levels of 2,000 cpm, considered to be the limit of background responses. Values less than 100 cpm are approximated to 100. A vaccinee is considered to be a responder to the antigen tested if the symbol is above the doubling level (diagonal broken line) as well as above the background level (2,000 cpm; horizontal line).
Figure Legend Snippet: Comparisons of T-cell proliferation (cpm) in response to M. bovis BCG as a control antigen (20 or 4 μg/ml) and PHA (5 μg/ml) in 12 volunteers before and after intranasal immunizations with a meningococcal OMV vaccine without adjuvant. Each symbol represents the preimmunization cpm ( x value) and the maximum postimmunization cpm ( y value) obtained for one vaccinee. The diagonal solid line in each graph indicates the same post- as preimmunization values, and the two broken lines indicate twofold increases and decreases. The dotted horizontal lines define postimmunization levels of 2,000 cpm, considered to be the limit of background responses. Values less than 100 cpm are approximated to 100. A vaccinee is considered to be a responder to the antigen tested if the symbol is above the doubling level (diagonal broken line) as well as above the background level (2,000 cpm; horizontal line).

Techniques Used:

32) Product Images from "Lower CD28+ T cell proportions were associated with CMV-seropositivity in patients with Hashimoto's thyroiditis"

Article Title: Lower CD28+ T cell proportions were associated with CMV-seropositivity in patients with Hashimoto's thyroiditis

Journal: BMC Endocrine Disorders

doi: 10.1186/1472-6823-13-34

Interferon-gamma-producing spot-forming-units (SFU) per 10 6 cells in patients with Hashimoto’s Thyroiditis (HT) and healthy controls (HC). Peripheral blood mononuclear cells (PBMCs) were either stimulated with PHA (A) or with CMV pp65 antigen (B) . A representative example of ELISPOT analysis showed similar results for HT patients and HC (C) . Negative controls were unstimulated. Experiments were performed in duplicates.
Figure Legend Snippet: Interferon-gamma-producing spot-forming-units (SFU) per 10 6 cells in patients with Hashimoto’s Thyroiditis (HT) and healthy controls (HC). Peripheral blood mononuclear cells (PBMCs) were either stimulated with PHA (A) or with CMV pp65 antigen (B) . A representative example of ELISPOT analysis showed similar results for HT patients and HC (C) . Negative controls were unstimulated. Experiments were performed in duplicates.

Techniques Used: Enzyme-linked Immunospot

33) Product Images from "Estrogen receptor α/HDAC/NFAT axis for delphinidin effects on proliferation and differentiation of T lymphocytes from patients with cardiovascular risks"

Article Title: Estrogen receptor α/HDAC/NFAT axis for delphinidin effects on proliferation and differentiation of T lymphocytes from patients with cardiovascular risks

Journal: Scientific Reports

doi: 10.1038/s41598-017-09933-4

Delphinidin inhibits T lymphocyte proliferation through ERα-dependent mechanism. ( A ) Histograms show the percentage of proliferation of cells exposed to either 10 −2 g/L of delphinidin (Del), 5 µg/mL PHA or both in presence or in absence of nonselective estrogen receptor antagonist (Fulvestrant 100 nM) for 48 h. Data are the mean ± SEM ( n = 8). ( B ) Western blot of phosphorylated ERK1/2 in T cells exposed to either 10 −2 g/L of delphinidin, 5 µg/mL PHA or both for 24 h in presence of fulvestrant. Histograms show densitometric analysis of phosphorylated ERK1/2 normalized to total ERK1/2 expression, Data are the mean ± SEM ( n = 6). ( C ) Representative traces ( left ) and histograms ( right ) showing the effect of 10 −2 g/L of delphinidin alone or after activation by 1 µM thapsigargine (TG) on [Ca 2+ ] i in Ca 2+ -containing PSS in presence of Ful (100 nM). ( D ) Representative traces ( left ) and histograms ( right ) showing the effect of delphinidin on [Ca 2+ ] i increase induced by 1.25 mM of CaCl 2 after depletion of intracellular stores in Ca 2+ -free PSS by thapsigargin in presence of fulvestrant. Data are the mean ± SEM ( n = 7). ( E ) Histograms show the percentage of proliferation of cells isolated from ERα WT or KO mice exposed to either 10 −2 g/L of delphinidin, 5 µg/mL PHA or both for 48 h. ( F ) Representative traces ( left ) and histograms ( right ) showing the effect of 10 −2 g/L of delphinidin (Del) alone or after activation by 1 µM thapsigargin (TG) on [Ca 2+ ] i in Ca 2+ -containing PSS of cells isolated from ERα WT mice. ( G ) Same experiments on cells isolated from ERα KO mice. Data are the mean ± SEM ( n = 4). * P
Figure Legend Snippet: Delphinidin inhibits T lymphocyte proliferation through ERα-dependent mechanism. ( A ) Histograms show the percentage of proliferation of cells exposed to either 10 −2 g/L of delphinidin (Del), 5 µg/mL PHA or both in presence or in absence of nonselective estrogen receptor antagonist (Fulvestrant 100 nM) for 48 h. Data are the mean ± SEM ( n = 8). ( B ) Western blot of phosphorylated ERK1/2 in T cells exposed to either 10 −2 g/L of delphinidin, 5 µg/mL PHA or both for 24 h in presence of fulvestrant. Histograms show densitometric analysis of phosphorylated ERK1/2 normalized to total ERK1/2 expression, Data are the mean ± SEM ( n = 6). ( C ) Representative traces ( left ) and histograms ( right ) showing the effect of 10 −2 g/L of delphinidin alone or after activation by 1 µM thapsigargine (TG) on [Ca 2+ ] i in Ca 2+ -containing PSS in presence of Ful (100 nM). ( D ) Representative traces ( left ) and histograms ( right ) showing the effect of delphinidin on [Ca 2+ ] i increase induced by 1.25 mM of CaCl 2 after depletion of intracellular stores in Ca 2+ -free PSS by thapsigargin in presence of fulvestrant. Data are the mean ± SEM ( n = 7). ( E ) Histograms show the percentage of proliferation of cells isolated from ERα WT or KO mice exposed to either 10 −2 g/L of delphinidin, 5 µg/mL PHA or both for 48 h. ( F ) Representative traces ( left ) and histograms ( right ) showing the effect of 10 −2 g/L of delphinidin (Del) alone or after activation by 1 µM thapsigargin (TG) on [Ca 2+ ] i in Ca 2+ -containing PSS of cells isolated from ERα WT mice. ( G ) Same experiments on cells isolated from ERα KO mice. Data are the mean ± SEM ( n = 4). * P

Techniques Used: Western Blot, Expressing, Activation Assay, Isolation, Mouse Assay

Delphinidin inhibits T lymphocyte proliferation through SOCE-dependent pathway. ( A ) Histograms show the percentage of proliferation of cells exposed to either 10 −2 g/L of delphinidin (Del), 5 µg/mL PHA or both, in absence or in presence of 10 µM of SOCE inhibitor (SKF96365) for 48 h. ( B ) Representative traces ( left ) and histograms ( right ) showing the effect of 10 −2 g/L of delphinidin (Del) alone or after activation by 1 µM thapsigargin (TG) on [Ca 2+ ] i in Ca 2+ -containing PSS. ( C ) Same experiments in presence of 10 µM of SOCE inhibitor (SKF96365). ( D ) Representative traces ( left ) and histograms ( right ) showing the effect of 10 −2 g/L delphinidin on [Ca 2+ ] i increase induced by 1.25 mM of CaCl 2 after depletion of intracellular stores in Ca 2+ -free PSS by 1 µM TG. ( E ) Same experiments in presence of 10 µM of SOCE inhibitor, SKF96365. Data are the mean ± SEM ( n = 7–11). * P
Figure Legend Snippet: Delphinidin inhibits T lymphocyte proliferation through SOCE-dependent pathway. ( A ) Histograms show the percentage of proliferation of cells exposed to either 10 −2 g/L of delphinidin (Del), 5 µg/mL PHA or both, in absence or in presence of 10 µM of SOCE inhibitor (SKF96365) for 48 h. ( B ) Representative traces ( left ) and histograms ( right ) showing the effect of 10 −2 g/L of delphinidin (Del) alone or after activation by 1 µM thapsigargin (TG) on [Ca 2+ ] i in Ca 2+ -containing PSS. ( C ) Same experiments in presence of 10 µM of SOCE inhibitor (SKF96365). ( D ) Representative traces ( left ) and histograms ( right ) showing the effect of 10 −2 g/L delphinidin on [Ca 2+ ] i increase induced by 1.25 mM of CaCl 2 after depletion of intracellular stores in Ca 2+ -free PSS by 1 µM TG. ( E ) Same experiments in presence of 10 µM of SOCE inhibitor, SKF96365. Data are the mean ± SEM ( n = 7–11). * P

Techniques Used: Activation Assay

Effect of delphinidin on the differentiation of T lymphocytes from healthy subjects. ( A ) Cells were stimulated for 24 h with 10 −2 g/L of delphinidin (Del), 5 µg/mL of PHA or both and stained for T-bet, GATA3, RORγt and FOXP3 transcription factors. Histograms show the percentage of positive cells for T-bet, GATA3, RORγt and FOXP3 expressions. Data are the mean ± SEM ( n = 7). ( B ) T cells were stimulated for 5 h with 10 −2 g/L of delphinidin (Del), 50 ng/mL phorbol-12-myristate-13-acetate (PMA) plus 1 µg/mL ionomycin (Ion) or both, in the presence of 5 µg/mL brefeldin A (BFA) for the last 3 h of culture and stained for IL-2, IFNγ, IL-4, IL-17A and IL-10 cytokines. Histograms show the MFI of positive cells for IL-2, IFNγ, IL-4, IL-17A, and IL-10, respectively. Data are the mean ± SEM ( n = 7). * P
Figure Legend Snippet: Effect of delphinidin on the differentiation of T lymphocytes from healthy subjects. ( A ) Cells were stimulated for 24 h with 10 −2 g/L of delphinidin (Del), 5 µg/mL of PHA or both and stained for T-bet, GATA3, RORγt and FOXP3 transcription factors. Histograms show the percentage of positive cells for T-bet, GATA3, RORγt and FOXP3 expressions. Data are the mean ± SEM ( n = 7). ( B ) T cells were stimulated for 5 h with 10 −2 g/L of delphinidin (Del), 50 ng/mL phorbol-12-myristate-13-acetate (PMA) plus 1 µg/mL ionomycin (Ion) or both, in the presence of 5 µg/mL brefeldin A (BFA) for the last 3 h of culture and stained for IL-2, IFNγ, IL-4, IL-17A and IL-10 cytokines. Histograms show the MFI of positive cells for IL-2, IFNγ, IL-4, IL-17A, and IL-10, respectively. Data are the mean ± SEM ( n = 7). * P

Techniques Used: Staining

Effects of delphinidin on proliferation, cell cycle progression and ERK1/2 pathway activation of T lymphocytes from healthy subjects. Histograms show the percentage of proliferation of cells exposed to T cell activators [10 µg/mL anti-CD3 plus 5 µg/mL anti-CD28, 5 µg/mL PHA or 10 ng/mL PMA plus 1 µM ionomycin (Ion)] in absence or in presence of 10 −2 g/L of delphinidin (Del) for 24 h ( A ) or 48 h ( B ). Data are the mean ± SEM ( n = 5–10). * P
Figure Legend Snippet: Effects of delphinidin on proliferation, cell cycle progression and ERK1/2 pathway activation of T lymphocytes from healthy subjects. Histograms show the percentage of proliferation of cells exposed to T cell activators [10 µg/mL anti-CD3 plus 5 µg/mL anti-CD28, 5 µg/mL PHA or 10 ng/mL PMA plus 1 µM ionomycin (Ion)] in absence or in presence of 10 −2 g/L of delphinidin (Del) for 24 h ( A ) or 48 h ( B ). Data are the mean ± SEM ( n = 5–10). * P

Techniques Used: Activation Assay

Effects of delphinidin on proliferation and Ca 2+ signaling of T lymphocytes from non-MetS and MetS patients. ( A ) Histograms show the percentage of proliferation of cells isolated from non-MetS patients exposed to either 10 −2 g/L of delphinidin (Del), 5 µg/mL PHA or both for 24 and 48 h. ( B ) Same experiments on cells isolated from MetS patients. Data are the mean ± SEM ( n = 8–31). ( C ) Representative traces ( left ) and histograms ( right ) showing the mean of the responses induced by 10 −2 g/L of delphinidin (Del) alone or after activation by 1 µM thapsigargin (TG) on [Ca 2+ ] i in Ca 2+ -containing PSS of T cells isolated from non-MetS patients. ( D ) Representative traces ( left ) and histogram ( right ) showing the effect of 10 −2 g/L Del on [Ca 2+ ] i increase induced by 1.25 mM of CaCl 2 after depletion of intracellular stores in Ca 2+ -free PSS by 1 µM TG of cells isolated from non-MetS patients. Data are the mean ± SEM ( n = 12–14). ( E and F ) Same experiments as ( C and D , respectively) on cells isolated from MetS patients. ( G ) Negative correlation between Del effect on PHA-induced proliferation and number of MetS criteria. Data are the mean ± SEM ( n = 16). * P
Figure Legend Snippet: Effects of delphinidin on proliferation and Ca 2+ signaling of T lymphocytes from non-MetS and MetS patients. ( A ) Histograms show the percentage of proliferation of cells isolated from non-MetS patients exposed to either 10 −2 g/L of delphinidin (Del), 5 µg/mL PHA or both for 24 and 48 h. ( B ) Same experiments on cells isolated from MetS patients. Data are the mean ± SEM ( n = 8–31). ( C ) Representative traces ( left ) and histograms ( right ) showing the mean of the responses induced by 10 −2 g/L of delphinidin (Del) alone or after activation by 1 µM thapsigargin (TG) on [Ca 2+ ] i in Ca 2+ -containing PSS of T cells isolated from non-MetS patients. ( D ) Representative traces ( left ) and histogram ( right ) showing the effect of 10 −2 g/L Del on [Ca 2+ ] i increase induced by 1.25 mM of CaCl 2 after depletion of intracellular stores in Ca 2+ -free PSS by 1 µM TG of cells isolated from non-MetS patients. Data are the mean ± SEM ( n = 12–14). ( E and F ) Same experiments as ( C and D , respectively) on cells isolated from MetS patients. ( G ) Negative correlation between Del effect on PHA-induced proliferation and number of MetS criteria. Data are the mean ± SEM ( n = 16). * P

Techniques Used: Isolation, Activation Assay

34) Product Images from "Proteasomal Inhibition Potentiates Latent HIV Reactivation"

Article Title: Proteasomal Inhibition Potentiates Latent HIV Reactivation

Journal: AIDS Research and Human Retroviruses

doi: 10.1089/aid.2020.0040

(A) Bortezomib does not activate uninfected CD + T cells. Uninfected human primary CD4+ T cells were stimulated for 24 h with DMSO, PMA and PHA, MG132, or bortezomib at the indicated concentrations. Cells were stained with anti-CD69 antibody and measured by FACS. (B) Percent viability was estimated using forward/side scatter and the percentage of live lymphocytes in 10,000 total cells analyzed. Data are presented with viability of untreated cells set to 100%. Experiments were repeated with three independent donors, each with three technical repeats. A representative experiment, from a single donor, is presented. Error bars represent standard error of the mean (*** p
Figure Legend Snippet: (A) Bortezomib does not activate uninfected CD + T cells. Uninfected human primary CD4+ T cells were stimulated for 24 h with DMSO, PMA and PHA, MG132, or bortezomib at the indicated concentrations. Cells were stained with anti-CD69 antibody and measured by FACS. (B) Percent viability was estimated using forward/side scatter and the percentage of live lymphocytes in 10,000 total cells analyzed. Data are presented with viability of untreated cells set to 100%. Experiments were repeated with three independent donors, each with three technical repeats. A representative experiment, from a single donor, is presented. Error bars represent standard error of the mean (*** p

Techniques Used: Staining, FACS

Uninfected cells were maintained in the same conditions. At 12 days postinfection, cells were stimulated for 24 h with DMSO, PMA and PHA, MG132, and bortezomib at the indicated concentrations. Cells were stained with anti-HSA antibody and measured by FACS. Data are presented as the percentage of cells positive for HSA expression. Experiments were repeated with three independent donors, each with three technical repeats. A representative experiment from a single donor is presented. Error bars represent standard error of the mean (*** p
Figure Legend Snippet: Uninfected cells were maintained in the same conditions. At 12 days postinfection, cells were stimulated for 24 h with DMSO, PMA and PHA, MG132, and bortezomib at the indicated concentrations. Cells were stained with anti-HSA antibody and measured by FACS. Data are presented as the percentage of cells positive for HSA expression. Experiments were repeated with three independent donors, each with three technical repeats. A representative experiment from a single donor is presented. Error bars represent standard error of the mean (*** p

Techniques Used: Staining, FACS, Expressing

Bortezomib increases Cyclin T1 and activates NF-κB. (A) Human PBMCs were stimulated for 24 h with DMSO and bortezomib. Nuclear extracts were prepared and run on 10% SDS-PAGE. Membranes were probed for anti-human CycT1 and β-actin. Densitometry was performed and normalized to β-actin, and the expression in untreated controls was set to 1. All samples represented were run on the same gel (space indicates lanes omitted from the figure). (B) Human PBMCs were stimulated for 9 h with DMSO, PMA and PHA, MG132, and bortezomib at indicated concentrations. Nuclear extracts were prepared and run on 10% SDS-PAGE. Membranes were probed for anti-human ph-IκBα, total IκBα, and β-actin. Densitometry was performed and normalized to total IκBα and β-actin, and the expression in untreated controls was set to 1. (C) Human PBMCs were stimulated for 1 h with DMSO, PMA and PHA, MG132, TNFα, and bortezomib at indicated concentrations. Nuclear extracts were prepared and run on 10% SDS-PAGE. Membranes were probed for ph-p65, total p65, and β-actin. Densitometry was performed and normalized to total p65 and β-actin, and the expression in untreated controls was set to 1. Results are representative of Western blots from three healthy donors. NF-κB, nuclear factor κB; PBMCs, peripheral blood mononuclear cells; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; TNFα, tumor necrosis factor alpha.
Figure Legend Snippet: Bortezomib increases Cyclin T1 and activates NF-κB. (A) Human PBMCs were stimulated for 24 h with DMSO and bortezomib. Nuclear extracts were prepared and run on 10% SDS-PAGE. Membranes were probed for anti-human CycT1 and β-actin. Densitometry was performed and normalized to β-actin, and the expression in untreated controls was set to 1. All samples represented were run on the same gel (space indicates lanes omitted from the figure). (B) Human PBMCs were stimulated for 9 h with DMSO, PMA and PHA, MG132, and bortezomib at indicated concentrations. Nuclear extracts were prepared and run on 10% SDS-PAGE. Membranes were probed for anti-human ph-IκBα, total IκBα, and β-actin. Densitometry was performed and normalized to total IκBα and β-actin, and the expression in untreated controls was set to 1. (C) Human PBMCs were stimulated for 1 h with DMSO, PMA and PHA, MG132, TNFα, and bortezomib at indicated concentrations. Nuclear extracts were prepared and run on 10% SDS-PAGE. Membranes were probed for ph-p65, total p65, and β-actin. Densitometry was performed and normalized to total p65 and β-actin, and the expression in untreated controls was set to 1. Results are representative of Western blots from three healthy donors. NF-κB, nuclear factor κB; PBMCs, peripheral blood mononuclear cells; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; TNFα, tumor necrosis factor alpha.

Techniques Used: SDS Page, Expressing, Western Blot, Polyacrylamide Gel Electrophoresis

35) Product Images from "HTLV-1 bZIP Factor Enhances T-Cell Proliferation by Impeding the Suppressive Signaling of Co-inhibitory Receptors"

Article Title: HTLV-1 bZIP Factor Enhances T-Cell Proliferation by Impeding the Suppressive Signaling of Co-inhibitory Receptors

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1006120

Expression of TIGIT, PD-1, BTLA and Lair-1 in ATL cases. (A) Relative mRNA expression levels of co-inhibitory receptors in resting healthy donor CD4 + T cells (n = 5), PHA-activated CD4 + T cells (n = 5) and ATL cells (n = 14) were evaluated by real-time RT-PCR. (B) Cells from the same groups were stained with anti-CD4, TIGIT, PD-1, BTLA and LAIR-1 antibodies. Expression of co-inhibitory receptors on CD4 + T cells was analyzed by flow cytometry. (C) The MFI of TIGIT, PD-1, BTLA and LAIR-1 on the cells are shown.
Figure Legend Snippet: Expression of TIGIT, PD-1, BTLA and Lair-1 in ATL cases. (A) Relative mRNA expression levels of co-inhibitory receptors in resting healthy donor CD4 + T cells (n = 5), PHA-activated CD4 + T cells (n = 5) and ATL cells (n = 14) were evaluated by real-time RT-PCR. (B) Cells from the same groups were stained with anti-CD4, TIGIT, PD-1, BTLA and LAIR-1 antibodies. Expression of co-inhibitory receptors on CD4 + T cells was analyzed by flow cytometry. (C) The MFI of TIGIT, PD-1, BTLA and LAIR-1 on the cells are shown.

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

36) Product Images from "Evaluation of optimum conditions for pachyman encapsulated in poly(d,l-lactic acid) nanospheres by response surface methodology and results of a related in vitro study"

Article Title: Evaluation of optimum conditions for pachyman encapsulated in poly(d,l-lactic acid) nanospheres by response surface methodology and results of a related in vitro study

Journal: International Journal of Nanomedicine

doi: 10.2147/IJN.S115742

Freshly prepared lymphocytes were seeded in transparent plates, incubated with PHYP, PLA, or PHY solutions at different concentrations, and cultured at 37°C in a humidified atmosphere with 5% CO 2 for 48 hours. Notes: ( A ) The cytotoxic activity of different nanosphere formulations on lymphocytes was measured by MTT assay and calculated as cell viability (% of control). ( B and C ) Lymphocyte proliferation assay on synergistic stimulation with PHA ( A 570 ) (a–e) ( B ) and LPS ( A 570 ) (a–d) ( C ) was also evaluated by MTT assay. Bars in the figure with different letters were significantly different ( P
Figure Legend Snippet: Freshly prepared lymphocytes were seeded in transparent plates, incubated with PHYP, PLA, or PHY solutions at different concentrations, and cultured at 37°C in a humidified atmosphere with 5% CO 2 for 48 hours. Notes: ( A ) The cytotoxic activity of different nanosphere formulations on lymphocytes was measured by MTT assay and calculated as cell viability (% of control). ( B and C ) Lymphocyte proliferation assay on synergistic stimulation with PHA ( A 570 ) (a–e) ( B ) and LPS ( A 570 ) (a–d) ( C ) was also evaluated by MTT assay. Bars in the figure with different letters were significantly different ( P

Techniques Used: Incubation, Proximity Ligation Assay, Cell Culture, Activity Assay, MTT Assay, Lymphocyte Proliferation Assay

37) Product Images from "Modulation of the Human T Cell Response by a Novel Non-Mitogenic Anti-CD3 Antibody"

Article Title: Modulation of the Human T Cell Response by a Novel Non-Mitogenic Anti-CD3 Antibody

Journal: PLoS ONE

doi: 10.1371/journal.pone.0094324

20-2b2 inhibited the activation of T cells in vitro . (A–C) The PBMCs of healthy donors were cultured with an anti-CD3 Ab (OKT3) (A), PHA (B), or PWM (C) in the presence (red symbols) or absence (black) of 20-2b2 (50% SN). (D) PBMCs were pre-treated with medium (black) or 20-2b2 (red) at 37°C. Cells were washed three times and cultured with PHA two hours later. Cell proliferation was measured two days later. Values are shown as the mean ± SD of triplicate cultures. Data are representative of more than three (A–C) and three (D) independent experiments. * p
Figure Legend Snippet: 20-2b2 inhibited the activation of T cells in vitro . (A–C) The PBMCs of healthy donors were cultured with an anti-CD3 Ab (OKT3) (A), PHA (B), or PWM (C) in the presence (red symbols) or absence (black) of 20-2b2 (50% SN). (D) PBMCs were pre-treated with medium (black) or 20-2b2 (red) at 37°C. Cells were washed three times and cultured with PHA two hours later. Cell proliferation was measured two days later. Values are shown as the mean ± SD of triplicate cultures. Data are representative of more than three (A–C) and three (D) independent experiments. * p

Techniques Used: Activation Assay, In Vitro, Cell Culture

38) Product Images from "Comparison of the oxidative respiratory burst and mitogen-induced leukocyte responses of camels, goats, sheep, and cows"

Article Title: Comparison of the oxidative respiratory burst and mitogen-induced leukocyte responses of camels, goats, sheep, and cows

Journal: Veterinary World

doi: 10.14202/vetworld.2022.1398-1407

Comparison stimulation indices of peripheral blood mononuclear leukocyte of camels, goats, sheep, and cows stimulated with concanavalin A, phytohemagglutinin and pokeweed mitogen.
Figure Legend Snippet: Comparison stimulation indices of peripheral blood mononuclear leukocyte of camels, goats, sheep, and cows stimulated with concanavalin A, phytohemagglutinin and pokeweed mitogen.

Techniques Used:

Comparison of stimulation indices of peripheral blood mononuclear leukocytes from camels, goats, sheep, and cows with phytohemagglutinin.
Figure Legend Snippet: Comparison of stimulation indices of peripheral blood mononuclear leukocytes from camels, goats, sheep, and cows with phytohemagglutinin.

Techniques Used:

39) Product Images from "Characterization of Simian Immunodeficiency Virus SIVSM/Human Immunodeficiency Virus Type 2 Vpx Function in Human Myeloid Cells ▿"

Article Title: Characterization of Simian Immunodeficiency Virus SIVSM/Human Immunodeficiency Virus Type 2 Vpx Function in Human Myeloid Cells ▿

Journal:

doi: 10.1128/JVI.01181-08

Vpx is required during the early steps of infection of myeloid blood cells and in differentiated THP-1 cells. Primary blood monocytes, macrophages, DCs, PHA/IL-2-stimulated PBLs, as well as cycling and differentiated THP-1 cells were compared for their
Figure Legend Snippet: Vpx is required during the early steps of infection of myeloid blood cells and in differentiated THP-1 cells. Primary blood monocytes, macrophages, DCs, PHA/IL-2-stimulated PBLs, as well as cycling and differentiated THP-1 cells were compared for their

Techniques Used: Infection

40) Product Images from "Retinoic acid and liver X receptor agonist synergistically inhibit HIV infection in CD4+ T cells by up-regulating ABCA1-mediated cholesterol efflux"

Article Title: Retinoic acid and liver X receptor agonist synergistically inhibit HIV infection in CD4+ T cells by up-regulating ABCA1-mediated cholesterol efflux

Journal: Lipids in Health and Disease

doi: 10.1186/1476-511X-11-69

ATRA induces ABCA1 expression in human primary CD4+ T cells. A ) Human CD4+ T cells isolated from healthy donors were primed with anti-CD3 and anti-CD28 antibodies and treated with DMSO or 1 μM of ATRA for 3 days. RNA isolation, reverse transcription, and real time PCR were done as described in Methods. ABCA1 RNA expression was normalized to GAPDH RNA expression, and relative RNA fold changes compared to that from DMSO treatment were plotted. B ) ABCA1 proteins were detected by Western blot with antibody specific to human ABCA1 as described. GAPDH was used as a loading control. C ) ABCA1 RNA expression in response to treatment with different concentrations of ATRA. D ) ABCA1 RNA expression in response to treatment with ATRA for various lengths of times. E ) CD4+ T cells were activated with anti-CD3 and anti-CD28 antibody or PMA and PHA and were also treated with either DMSO or ATRA. ABCA1 RNA level was analyzed by real-time PCR. F ) Anti-CD3/CD28 antibody primed CD4+ T cells were treated with DMSO or 10 μM of U0126 for 2 hours and then treated with or without ATRA. ABCA1 RNA level was quantitated by real-time PCR. Results are representative of three experiments with cells from three different donors.
Figure Legend Snippet: ATRA induces ABCA1 expression in human primary CD4+ T cells. A ) Human CD4+ T cells isolated from healthy donors were primed with anti-CD3 and anti-CD28 antibodies and treated with DMSO or 1 μM of ATRA for 3 days. RNA isolation, reverse transcription, and real time PCR were done as described in Methods. ABCA1 RNA expression was normalized to GAPDH RNA expression, and relative RNA fold changes compared to that from DMSO treatment were plotted. B ) ABCA1 proteins were detected by Western blot with antibody specific to human ABCA1 as described. GAPDH was used as a loading control. C ) ABCA1 RNA expression in response to treatment with different concentrations of ATRA. D ) ABCA1 RNA expression in response to treatment with ATRA for various lengths of times. E ) CD4+ T cells were activated with anti-CD3 and anti-CD28 antibody or PMA and PHA and were also treated with either DMSO or ATRA. ABCA1 RNA level was analyzed by real-time PCR. F ) Anti-CD3/CD28 antibody primed CD4+ T cells were treated with DMSO or 10 μM of U0126 for 2 hours and then treated with or without ATRA. ABCA1 RNA level was quantitated by real-time PCR. Results are representative of three experiments with cells from three different donors.

Techniques Used: Expressing, Isolation, Real-time Polymerase Chain Reaction, RNA Expression, Western Blot

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    Millipore pha
    MUC1 peptides couple to anti-human DNGR-1 can induce CD8 + T cells in vitro. BDCA-3 + cells were isolated from PBMCs and incubated with the LLL peptide coupled to anti-human DNGR-1 or isotype control, in the presence of anti-CD40 and poly-IC, for 1 hour before incubating with autologous T cells for 7 days. The cells were then pulsed with peptide following a further 7 days incubation stimulated with autologous monocyte-derived DCs pulsed with peptide. (A) Seven days later, the cells were pulsed for 4 hour with LLL peptide and IFN-γ secretion by CD8 + T cells measured by IFN-γ Secretion Assay. An aliquot of anti-DNGR-1-peptide stimulated cells was incubated for 4 hour with 5 μg/mL <t>PHA</t> and 1 μg/mL anti-human <t>CD3</t> (Control DNGR-1). Data shown are representative of four independent experiments. (B, C) Cells were also pulsed with (B) MCF-7 cells and (C) Capan-1 cells and IFN-γ production was measured by intracellular cytokine staining. An aliquot of either isotype control (Control Ig) or anti-DNGR-1 (Control DNGR-1) stimulated cells was incubated for 4 hour with PBMC:Dynabeads® human T-Expander CD3/CD28. All the cells were analysed after gating on live cells. (D, E) Expression of endogenous MUC1 by (D) Capan-1 cells and (E) MCF-7 cells was determined by HMFG2 binding (black-line histogram) as assessed by flow cytometry. (Filled histogram, isotype control.)
    Pha, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell surface expression of PDI and / or Thioredoxin. PM-1 T cells (panel A ), primary human MDM (panel B ) and PBLs (panel C ) were isolated and cultured as described in Methods. Subsequently, the cells were fixed with 1.6% paraformaldehyde and treated with NH 4 Cl to quench the reactive aldehyde groups before labeling. The PDI and Trx levels on the cell surface were detected by a combination of directly labeled anti-PDI-FITC and anti-Trx-APC mAbs from the same clones used in the HIV-1 infection experiments. In addition, the unstimulated and <t>PHA-P</t> stimulated PBLs were labeled with anti-CD4-Pacific blue conjugated mAb, and the PDI and Trx expression were analyzed on the CD4+ lymphocyte (Ly) population. The results were analyzed and the final graphs were created using FlowJoe software.
    Pha P, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MUC1 peptides couple to anti-human DNGR-1 can induce CD8 + T cells in vitro. BDCA-3 + cells were isolated from PBMCs and incubated with the LLL peptide coupled to anti-human DNGR-1 or isotype control, in the presence of anti-CD40 and poly-IC, for 1 hour before incubating with autologous T cells for 7 days. The cells were then pulsed with peptide following a further 7 days incubation stimulated with autologous monocyte-derived DCs pulsed with peptide. (A) Seven days later, the cells were pulsed for 4 hour with LLL peptide and IFN-γ secretion by CD8 + T cells measured by IFN-γ Secretion Assay. An aliquot of anti-DNGR-1-peptide stimulated cells was incubated for 4 hour with 5 μg/mL PHA and 1 μg/mL anti-human CD3 (Control DNGR-1). Data shown are representative of four independent experiments. (B, C) Cells were also pulsed with (B) MCF-7 cells and (C) Capan-1 cells and IFN-γ production was measured by intracellular cytokine staining. An aliquot of either isotype control (Control Ig) or anti-DNGR-1 (Control DNGR-1) stimulated cells was incubated for 4 hour with PBMC:Dynabeads® human T-Expander CD3/CD28. All the cells were analysed after gating on live cells. (D, E) Expression of endogenous MUC1 by (D) Capan-1 cells and (E) MCF-7 cells was determined by HMFG2 binding (black-line histogram) as assessed by flow cytometry. (Filled histogram, isotype control.)

    Journal: European Journal of Immunology

    Article Title: Targeting DNGR-1 (CLEC9A) with antibody/MUC1 peptide conjugates as a vaccine for carcinomas

    doi: 10.1002/eji.201344076

    Figure Lengend Snippet: MUC1 peptides couple to anti-human DNGR-1 can induce CD8 + T cells in vitro. BDCA-3 + cells were isolated from PBMCs and incubated with the LLL peptide coupled to anti-human DNGR-1 or isotype control, in the presence of anti-CD40 and poly-IC, for 1 hour before incubating with autologous T cells for 7 days. The cells were then pulsed with peptide following a further 7 days incubation stimulated with autologous monocyte-derived DCs pulsed with peptide. (A) Seven days later, the cells were pulsed for 4 hour with LLL peptide and IFN-γ secretion by CD8 + T cells measured by IFN-γ Secretion Assay. An aliquot of anti-DNGR-1-peptide stimulated cells was incubated for 4 hour with 5 μg/mL PHA and 1 μg/mL anti-human CD3 (Control DNGR-1). Data shown are representative of four independent experiments. (B, C) Cells were also pulsed with (B) MCF-7 cells and (C) Capan-1 cells and IFN-γ production was measured by intracellular cytokine staining. An aliquot of either isotype control (Control Ig) or anti-DNGR-1 (Control DNGR-1) stimulated cells was incubated for 4 hour with PBMC:Dynabeads® human T-Expander CD3/CD28. All the cells were analysed after gating on live cells. (D, E) Expression of endogenous MUC1 by (D) Capan-1 cells and (E) MCF-7 cells was determined by HMFG2 binding (black-line histogram) as assessed by flow cytometry. (Filled histogram, isotype control.)

    Article Snippet: As a positive control for the assay, aliquots of T cells were incubated with a mixture of 5 μg/mL PHA (Sigma) and 1 μg/mL anti-human CD3 (clone UCHT1; Millipore).

    Techniques: In Vitro, Isolation, Incubation, Derivative Assay, Staining, Expressing, Binding Assay, Flow Cytometry, Cytometry

    uc002yug.2 increases viral replication and reactivation in CD4 + T cells. (A and B) uc002yug.2 was correlated with HIV-1 loads in plasma of HIV-1-infected patients. (A) Expression levels of uc002yug.2 in CD4 + T cells isolated from HIV-1 patients who underwent HAART ( n = 12) and HIV-1 patients who had not received HAART ( n = 12) were detected by qRT-PCR. The geometric means of the β-actin, GAPDH, and HMBS genes were used for normalization. (B) The HIV-1 loads and uc002yug.2 RNA levels of HIV-1-infected patients who had not received HAART were plotted, and linear regression analysis was performed. The geometric means of the β-actin, GAPDH, and HMBS genes were used for normalization. (C) uc002yug.2 increases viral replication. Primary CD4 + T lymphocytes from healthy donors were nucleofected with uc002yug.2 or control vector and then were infected with HIV-1 NL4-3. HIV-1 production in the supernatant was quantified with p24 ELISA at indicated time points postinfection. (D) uc002yug.2 increases HIV-1 reactivation in primary resting CD4 + T cells from patients. Resting CD4 + T cells were isolated from HAART-treated patients and nucleofected with uc002yug.2 or control vector. P1 to P3 represent three patients. HIV-1 reactivation in CD4 + T cells upon PHA-M (5 ng/ml) stimulation was detected by measuring p24 levels in the supernatants by ELISA.

    Journal: Journal of Virology

    Article Title: Long Noncoding RNA uc002yug.2 Activates HIV-1 Latency through Regulation of mRNA Levels of Various RUNX1 Isoforms and Increased Tat Expression

    doi: 10.1128/JVI.01844-17

    Figure Lengend Snippet: uc002yug.2 increases viral replication and reactivation in CD4 + T cells. (A and B) uc002yug.2 was correlated with HIV-1 loads in plasma of HIV-1-infected patients. (A) Expression levels of uc002yug.2 in CD4 + T cells isolated from HIV-1 patients who underwent HAART ( n = 12) and HIV-1 patients who had not received HAART ( n = 12) were detected by qRT-PCR. The geometric means of the β-actin, GAPDH, and HMBS genes were used for normalization. (B) The HIV-1 loads and uc002yug.2 RNA levels of HIV-1-infected patients who had not received HAART were plotted, and linear regression analysis was performed. The geometric means of the β-actin, GAPDH, and HMBS genes were used for normalization. (C) uc002yug.2 increases viral replication. Primary CD4 + T lymphocytes from healthy donors were nucleofected with uc002yug.2 or control vector and then were infected with HIV-1 NL4-3. HIV-1 production in the supernatant was quantified with p24 ELISA at indicated time points postinfection. (D) uc002yug.2 increases HIV-1 reactivation in primary resting CD4 + T cells from patients. Resting CD4 + T cells were isolated from HAART-treated patients and nucleofected with uc002yug.2 or control vector. P1 to P3 represent three patients. HIV-1 reactivation in CD4 + T cells upon PHA-M (5 ng/ml) stimulation was detected by measuring p24 levels in the supernatants by ELISA.

    Article Snippet: Cells were collected for qRT-PCR or stimulated with phytohemagglutinin M (PHA-M) (5 ng/ml; Sigma-Aldrich) for 7 days.

    Techniques: Infection, Expressing, Isolation, Quantitative RT-PCR, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

    A . Representative electrophoresis (left) and densitometry quantification of REL cDNA from EBV-transformed lymphoblastoid cell lines (EBV-LCLs) from controls and the patient. B. Schematic of REL cDNA and protein with the patient’s deleted nucleotides and residues in red. C . Representative immunoblot of c-Rel in lysates of peripheral blood mononuclear cells (PBMCs) from the patient and controls. D . CD3 + T cell proliferation to phytohemagglutinin (PHA), measured by CellTrace Violet (CTV) dilution. E . IL-2 secretion from patient and control PBMCs, with or without PHA stimulation. F . CD3 + T cell proliferation to PHA+IL-2. G, H . Secretion of IFN-γ ( G ) and IL-23 ( H ) by PBMCs from the patient and controls cultured with the indicated stimuli. I, J . Secretion of IL-12 ( I ) and IL-8 ( J ) by EBV-LCLs from the patient and controls stimulated with PDBu. K . CD19 + B cell proliferation to anti-CD40+IL-21. L . IL-21 secretion by PHA blasts from the patient or controls with or without PHA+IL-23 stimulation. M . Schematic of the contribution of c-Rel to adaptive and innate immunity. *p

    Journal: The Journal of allergy and clinical immunology

    Article Title: Combined immunodeficiency in a patient with deficiency of c-Rel

    doi: 10.1016/j.jaci.2019.05.003

    Figure Lengend Snippet: A . Representative electrophoresis (left) and densitometry quantification of REL cDNA from EBV-transformed lymphoblastoid cell lines (EBV-LCLs) from controls and the patient. B. Schematic of REL cDNA and protein with the patient’s deleted nucleotides and residues in red. C . Representative immunoblot of c-Rel in lysates of peripheral blood mononuclear cells (PBMCs) from the patient and controls. D . CD3 + T cell proliferation to phytohemagglutinin (PHA), measured by CellTrace Violet (CTV) dilution. E . IL-2 secretion from patient and control PBMCs, with or without PHA stimulation. F . CD3 + T cell proliferation to PHA+IL-2. G, H . Secretion of IFN-γ ( G ) and IL-23 ( H ) by PBMCs from the patient and controls cultured with the indicated stimuli. I, J . Secretion of IL-12 ( I ) and IL-8 ( J ) by EBV-LCLs from the patient and controls stimulated with PDBu. K . CD19 + B cell proliferation to anti-CD40+IL-21. L . IL-21 secretion by PHA blasts from the patient or controls with or without PHA+IL-23 stimulation. M . Schematic of the contribution of c-Rel to adaptive and innate immunity. *p

    Article Snippet: T cell proliferation was quantified using dilution of CellTrace™ Violet Cell Proliferation Kit (Invitrogen, Waltham, MA, USA) after stimulation with PHA (5 μg/mL; Sigma Aldrich, St. Louis, MO, USA) ± IL-2 (40 U/mL; R & D Systems, Minneapolis, MN, USA) for 2.5 days, at which point cell culture supernatants were also harvested for measure of IFN-γ and IL-2.

    Techniques: Electrophoresis, Transformation Assay, Cell Culture

    Cell surface expression of PDI and / or Thioredoxin. PM-1 T cells (panel A ), primary human MDM (panel B ) and PBLs (panel C ) were isolated and cultured as described in Methods. Subsequently, the cells were fixed with 1.6% paraformaldehyde and treated with NH 4 Cl to quench the reactive aldehyde groups before labeling. The PDI and Trx levels on the cell surface were detected by a combination of directly labeled anti-PDI-FITC and anti-Trx-APC mAbs from the same clones used in the HIV-1 infection experiments. In addition, the unstimulated and PHA-P stimulated PBLs were labeled with anti-CD4-Pacific blue conjugated mAb, and the PDI and Trx expression were analyzed on the CD4+ lymphocyte (Ly) population. The results were analyzed and the final graphs were created using FlowJoe software.

    Journal: Retrovirology

    Article Title: Cell-type specific requirements for thiol/disulfide exchange during HIV-1 entry and infection

    doi: 10.1186/1742-4690-9-97

    Figure Lengend Snippet: Cell surface expression of PDI and / or Thioredoxin. PM-1 T cells (panel A ), primary human MDM (panel B ) and PBLs (panel C ) were isolated and cultured as described in Methods. Subsequently, the cells were fixed with 1.6% paraformaldehyde and treated with NH 4 Cl to quench the reactive aldehyde groups before labeling. The PDI and Trx levels on the cell surface were detected by a combination of directly labeled anti-PDI-FITC and anti-Trx-APC mAbs from the same clones used in the HIV-1 infection experiments. In addition, the unstimulated and PHA-P stimulated PBLs were labeled with anti-CD4-Pacific blue conjugated mAb, and the PDI and Trx expression were analyzed on the CD4+ lymphocyte (Ly) population. The results were analyzed and the final graphs were created using FlowJoe software.

    Article Snippet: After stimulation for 72 h with PHA-P 10 μg/ml (Sigma-Aldrich, St. Louis, MO), PBL were washed, counted and infected with HIV-1 Env pseudotyped or HIV-1 BlaM-containing virions in 24 well plates (106 cells/well).

    Techniques: Expressing, Isolation, Cell Culture, Labeling, Clone Assay, Infection, Software