Structured Review

Becton Dickinson monensin
KIR2DS4 is functional on dNK cells. pbNK and dNK cells from KIR2DS4 + donors were incubated in wells coated with anti-KIR2DS4 or an isotype control for 5 h in the presence of <t>monensin.</t> ( A ) dNK cells from a represenative donor, gated as in , are shown
Monensin, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Price from $9.99 to $1999.99
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86/100 stars

Images

1) Product Images from "Activating KIR2DS4 Is Expressed by Uterine NK Cells and Contributes to Successful Pregnancy"

Article Title: Activating KIR2DS4 Is Expressed by Uterine NK Cells and Contributes to Successful Pregnancy

Journal: The Journal of Immunology Author Choice

doi: 10.4049/jimmunol.1601279

KIR2DS4 is functional on dNK cells. pbNK and dNK cells from KIR2DS4 + donors were incubated in wells coated with anti-KIR2DS4 or an isotype control for 5 h in the presence of monensin. ( A ) dNK cells from a represenative donor, gated as in , are shown
Figure Legend Snippet: KIR2DS4 is functional on dNK cells. pbNK and dNK cells from KIR2DS4 + donors were incubated in wells coated with anti-KIR2DS4 or an isotype control for 5 h in the presence of monensin. ( A ) dNK cells from a represenative donor, gated as in , are shown

Techniques Used: Functional Assay, Incubation

2) Product Images from "NADPH Oxidase-2 Derived ROS Dictates Murine DC Cytokine-Mediated Cell Fate Decisions during CD4 T Helper-Cell Commitment"

Article Title: NADPH Oxidase-2 Derived ROS Dictates Murine DC Cytokine-Mediated Cell Fate Decisions during CD4 T Helper-Cell Commitment

Journal: PLoS ONE

doi: 10.1371/journal.pone.0028198

p47 phox−/− DC bias more OT-II T lymphocytes to secrete IFNγ. OT-II lymphocytes were stimulated with mature OVA 323–339 peptide-pulsed DC for 96 hours. (A) Secreted IFNγ detected in supernatants of 96 hour DC-OT-II co-cultures. The stimulated OT-II lymphocytes were rested in medium supplemented with rIL-2 for six days, and then restimulated with PMA and ionomycin for 4 hours, with monensin added for the last 2 hours. (B) Intracellular IFNγ production was then assessed by flow cytometry. Percentages of OT-II lymphocytes expressing IFNγ. The data are the mean (± SEM) percentage for 4 individual experiments with 3–4 of each genotype/experiment ** p
Figure Legend Snippet: p47 phox−/− DC bias more OT-II T lymphocytes to secrete IFNγ. OT-II lymphocytes were stimulated with mature OVA 323–339 peptide-pulsed DC for 96 hours. (A) Secreted IFNγ detected in supernatants of 96 hour DC-OT-II co-cultures. The stimulated OT-II lymphocytes were rested in medium supplemented with rIL-2 for six days, and then restimulated with PMA and ionomycin for 4 hours, with monensin added for the last 2 hours. (B) Intracellular IFNγ production was then assessed by flow cytometry. Percentages of OT-II lymphocytes expressing IFNγ. The data are the mean (± SEM) percentage for 4 individual experiments with 3–4 of each genotype/experiment ** p

Techniques Used: Flow Cytometry, Cytometry, Expressing

3) Product Images from "The Profile of T Cell Responses in Bacille Calmette–Guérin-Primed Mice Boosted by a Novel Sendai Virus Vectored Anti-Tuberculosis Vaccine"

Article Title: The Profile of T Cell Responses in Bacille Calmette–Guérin-Primed Mice Boosted by a Novel Sendai Virus Vectored Anti-Tuberculosis Vaccine

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.01796

Recall T cell responses against specific stimulation post Mycobacterium tuberculosis challenge determined by ICS assay. The immunization and infection schedule were described in Figure 6 A. Lung cells were stimulated ex vivo with Ag85AB peptides in the presence of monensin and brefeldin A and analyzed for cytokine production by ICS assay. T cells producing IFN-γ, IL-2, and TNF-α were analyzed and their percentage in CD4 + T cells (A) and CD8 + T cells (B) are shown. The percentage of seven subpopulations based on the production of three cytokines in any combination in the total CD4 + (C) and CD8 + (D) T cells and the pie chart analysis (E) are shown. Significant differences in frequency of T cell subsets are indicated. Data are representative of two independent experiments with three mice per group. * P
Figure Legend Snippet: Recall T cell responses against specific stimulation post Mycobacterium tuberculosis challenge determined by ICS assay. The immunization and infection schedule were described in Figure 6 A. Lung cells were stimulated ex vivo with Ag85AB peptides in the presence of monensin and brefeldin A and analyzed for cytokine production by ICS assay. T cells producing IFN-γ, IL-2, and TNF-α were analyzed and their percentage in CD4 + T cells (A) and CD8 + T cells (B) are shown. The percentage of seven subpopulations based on the production of three cytokines in any combination in the total CD4 + (C) and CD8 + (D) T cells and the pie chart analysis (E) are shown. Significant differences in frequency of T cell subsets are indicated. Data are representative of two independent experiments with three mice per group. * P

Techniques Used: Infection, Ex Vivo, Mouse Assay

Flow cytometric analysis of ICS in the splenocytes of vaccinated mice. Splenocytes were collected 4 weeks after the last inoculation, incubated with Ag85AB peptides (5 µg/ml) in the presence of monensin and brefeldin A, and analyzed for cytokine production by ICS assay. CD3 + CD4 + cells (CD4 + T cells) and CD3 + CD4 − cells (CD8 + T cells) producing IFN-γ, IL-2, and TNF-α were analyzed. Representative flow cytometric plots of intracellular staining in CD4 + T cells are shown in (A) , and summary data of single cytokine producing CD4 + T cells (B) and CD8 + T cells (C) are shown with significant differences indicated. Data are representative of two independent experiments with at least four mice per group. * P
Figure Legend Snippet: Flow cytometric analysis of ICS in the splenocytes of vaccinated mice. Splenocytes were collected 4 weeks after the last inoculation, incubated with Ag85AB peptides (5 µg/ml) in the presence of monensin and brefeldin A, and analyzed for cytokine production by ICS assay. CD3 + CD4 + cells (CD4 + T cells) and CD3 + CD4 − cells (CD8 + T cells) producing IFN-γ, IL-2, and TNF-α were analyzed. Representative flow cytometric plots of intracellular staining in CD4 + T cells are shown in (A) , and summary data of single cytokine producing CD4 + T cells (B) and CD8 + T cells (C) are shown with significant differences indicated. Data are representative of two independent experiments with at least four mice per group. * P

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

Related Articles

Incubation:

Article Title: Novel epitope evoking CD138 antigen-specific cytotoxic T lymphocytes targeting multiple myeloma and other plasma cell disorders
Article Snippet: .. After 1 h incubation, Brefeldin A (BD) and Monensin (BD) were added and the cells were incubated for an additional 4 h. Following incubation, the cells were then harvested, stained with CD8-PE mAb for 30 min at 4°C, and analysed by flow cytometry. .. The expression of CD107ab by the CD8+ T cells was determined as a measure of degranulation of the CD138 peptide-specific CD8+ CTL in response to target cells.

Article Title: Myeloma-specific multiple peptides able to generate cytotoxic T lymphocytes: A potential therapeutic application in multiple myeloma and other plasma cell disorders
Article Snippet: .. After 1 hour incubation, CD28/CD49d mAb (BD), as well as protein transport inhibitors Brefeldin A (BD) and Monensin (BD), were added to the cultures and incubated for an additional 5 hours. .. As a baseline control, MP-CTL were cultured in media with CD28/CD49d mAb, Brefeldin A, and Monensin alone.

Article Title: Therapy of lymphoma by immune checkpoint inhibitors: the role of T cells, NK cells and cytokine-induced tumor senescence
Article Snippet: PD-1 and IFN-γ expression of NK cells were then analyzed as described above. .. To determine NK-cell degranulation in the presence of NK-sensitive target cells, NK cells were highly enriched from λ-MYC spleens and again incubated with YAC-1 cells in a 1:1 ratio together with Brefeldin A, Monensin and PE-labeled anti-CD107a mAb (clone 1D4B; BD). .. After 5 hours, cells were counterstained with anti-NK1.1 and analyzed by FACS.

Staining:

Article Title: Novel epitope evoking CD138 antigen-specific cytotoxic T lymphocytes targeting multiple myeloma and other plasma cell disorders
Article Snippet: .. After 1 h incubation, Brefeldin A (BD) and Monensin (BD) were added and the cells were incubated for an additional 4 h. Following incubation, the cells were then harvested, stained with CD8-PE mAb for 30 min at 4°C, and analysed by flow cytometry. .. The expression of CD107ab by the CD8+ T cells was determined as a measure of degranulation of the CD138 peptide-specific CD8+ CTL in response to target cells.

Article Title: Friend retrovirus infection induces the development of memory-like natural killer cells
Article Snippet: Zombie UV Fixable Viability Kit (BioLegend) was used for the exclusion of dead cells. .. If stimulation was necessary, cells were stimulated with ionomycin (500 ng/ml), phorbol myristate acetate (PMA; 25 ng/ml), monensin (1X), and brefeldin A (2 µg/ml) diluted in Iscove’s modified Dulbecco’s medium (IMDM) buffer at 37 °C for 3 h. For intracellular stainings, cells were fixed with the Cytofix/Cytoperm Fixation/Permeabilization kit (BD) or with Foxp3/Transcription Factor staining buffer set (eBioscience). .. Cells were measured at LSR II (BD).

Article Title: Description of organ-specific phenotype, and functional characteristics of tissue resident lymphocytes from liver transplantation donor and research on immune tolerance mechanism of liver
Article Snippet: Adding anti-human CD3(16-0037-85, e-bioscience) (clone:OKT3), at a final concentration of 10 ug/ml to 100 ul PBS(0014217, BI) solution, and putting on a packet plate (352360, BD Falcon) and incubating for 3 hours at 37° C to spare; (2) Washing the plate twice with 200 ul cold PBS solution; putting mononuclear cells from peripheral blood, lymph node, spleen, and liver perfusion on the plates (each plate contains 1 million cells). (3) Adding RPMI 1640 medium, anti-human CD28 (clone:CD28) (16-0289-81, ebioscience) at a final concentration of 5 ug/ml and PHA (Phytohaemagglutinin) (L8754-50, sigma) at a final concentration of 5 ug/ml respectively, incubating for 12 hrs at 37° C in 5% CO2. .. With BD Golgistop protein transport inhibitor (554715, BD) containing monensin at a final concentration of 6 μg/ml, incubating for an additional 5 hrs at 37° C in 5% CO2. (4) After a total of 17 hours cell culture preparation, mononuclear cells were washed and stained with cell surface NK/CD56+ T cell markers, CD56-Pacific Blue and CD3-PECy(7557851, BD), T cells markers, CD8-Alexa FLOUR 488(557696, BD), and CD4-PECy5 (555348, BD) for 30 min. .. Samples were then fixed and permeabilized according to the manufacturer’s instructions and stained for intracellular IFN-γ-APC (554702, BD) and IL-10-PE (16-7108-85, ebioscience) for an additional 30 min. (5) After washing, cells were re-suspended in 1% paraformaldehyde (AR1068, Boster Wuhan) until five-color flow cytometric analysis was performed on a LSRII instrument (BD Biosciences).

Flow Cytometry:

Article Title: Novel epitope evoking CD138 antigen-specific cytotoxic T lymphocytes targeting multiple myeloma and other plasma cell disorders
Article Snippet: .. After 1 h incubation, Brefeldin A (BD) and Monensin (BD) were added and the cells were incubated for an additional 4 h. Following incubation, the cells were then harvested, stained with CD8-PE mAb for 30 min at 4°C, and analysed by flow cytometry. .. The expression of CD107ab by the CD8+ T cells was determined as a measure of degranulation of the CD138 peptide-specific CD8+ CTL in response to target cells.

Article Title: Activity of Uncleaved Caspase-8 Controls Anti-bacterial Immune Defense and TLR-Induced Cytokine Production Independent of Cell Death
Article Snippet: On days 3 and 10, lungs were harvested and homogenized in Trizol Reagent (Invitrogen/ThermoFisher) for RT-qPCR analysis. .. Flow cytometry For animal studies, mesenteric lymph nodes were isolated and plated in complete-DMEM containing brefeldin A (Sigma) and monensin (BD) in a 37°C humidified incubator for 5 hrs. ..

Cytometry:

Article Title: Novel epitope evoking CD138 antigen-specific cytotoxic T lymphocytes targeting multiple myeloma and other plasma cell disorders
Article Snippet: .. After 1 h incubation, Brefeldin A (BD) and Monensin (BD) were added and the cells were incubated for an additional 4 h. Following incubation, the cells were then harvested, stained with CD8-PE mAb for 30 min at 4°C, and analysed by flow cytometry. .. The expression of CD107ab by the CD8+ T cells was determined as a measure of degranulation of the CD138 peptide-specific CD8+ CTL in response to target cells.

Article Title: Activity of Uncleaved Caspase-8 Controls Anti-bacterial Immune Defense and TLR-Induced Cytokine Production Independent of Cell Death
Article Snippet: On days 3 and 10, lungs were harvested and homogenized in Trizol Reagent (Invitrogen/ThermoFisher) for RT-qPCR analysis. .. Flow cytometry For animal studies, mesenteric lymph nodes were isolated and plated in complete-DMEM containing brefeldin A (Sigma) and monensin (BD) in a 37°C humidified incubator for 5 hrs. ..

Isolation:

Article Title: Activity of Uncleaved Caspase-8 Controls Anti-bacterial Immune Defense and TLR-Induced Cytokine Production Independent of Cell Death
Article Snippet: On days 3 and 10, lungs were harvested and homogenized in Trizol Reagent (Invitrogen/ThermoFisher) for RT-qPCR analysis. .. Flow cytometry For animal studies, mesenteric lymph nodes were isolated and plated in complete-DMEM containing brefeldin A (Sigma) and monensin (BD) in a 37°C humidified incubator for 5 hrs. ..

Article Title: RIPK1-dependent apoptosis bypasses pathogen blockade of innate signaling to promote immune defense
Article Snippet: Flow cytometry MLN cells were isolated by passing through a 100-µM cell strainer. .. To measure cytokine expression by innate immune cells (neutrophils, monocytes, and DCs), 3–5 × 106 isolated cells were plated in a 96-well plate and cultured for 5 h in the presence of brefeldin A (Sigma) and monensin (BD) in a 37°C humidified incubator in complete DMEM containing penicillin–streptomycin and 2-mercaptoethanol. .. For stimulation and intracellular cytokine staining of T cells, 2–3 × 106 isolated cells were plated and cultured for 2 h in the presence of either heat-killed Yp (HKYp) or YopE69–77 peptide and for 3 h in the presence of brefeldin A and monensin.

Expressing:

Article Title: RIPK1-dependent apoptosis bypasses pathogen blockade of innate signaling to promote immune defense
Article Snippet: Flow cytometry MLN cells were isolated by passing through a 100-µM cell strainer. .. To measure cytokine expression by innate immune cells (neutrophils, monocytes, and DCs), 3–5 × 106 isolated cells were plated in a 96-well plate and cultured for 5 h in the presence of brefeldin A (Sigma) and monensin (BD) in a 37°C humidified incubator in complete DMEM containing penicillin–streptomycin and 2-mercaptoethanol. .. For stimulation and intracellular cytokine staining of T cells, 2–3 × 106 isolated cells were plated and cultured for 2 h in the presence of either heat-killed Yp (HKYp) or YopE69–77 peptide and for 3 h in the presence of brefeldin A and monensin.

Cell Culture:

Article Title: RIPK1-dependent apoptosis bypasses pathogen blockade of innate signaling to promote immune defense
Article Snippet: Flow cytometry MLN cells were isolated by passing through a 100-µM cell strainer. .. To measure cytokine expression by innate immune cells (neutrophils, monocytes, and DCs), 3–5 × 106 isolated cells were plated in a 96-well plate and cultured for 5 h in the presence of brefeldin A (Sigma) and monensin (BD) in a 37°C humidified incubator in complete DMEM containing penicillin–streptomycin and 2-mercaptoethanol. .. For stimulation and intracellular cytokine staining of T cells, 2–3 × 106 isolated cells were plated and cultured for 2 h in the presence of either heat-killed Yp (HKYp) or YopE69–77 peptide and for 3 h in the presence of brefeldin A and monensin.

Article Title: Description of organ-specific phenotype, and functional characteristics of tissue resident lymphocytes from liver transplantation donor and research on immune tolerance mechanism of liver
Article Snippet: Adding anti-human CD3(16-0037-85, e-bioscience) (clone:OKT3), at a final concentration of 10 ug/ml to 100 ul PBS(0014217, BI) solution, and putting on a packet plate (352360, BD Falcon) and incubating for 3 hours at 37° C to spare; (2) Washing the plate twice with 200 ul cold PBS solution; putting mononuclear cells from peripheral blood, lymph node, spleen, and liver perfusion on the plates (each plate contains 1 million cells). (3) Adding RPMI 1640 medium, anti-human CD28 (clone:CD28) (16-0289-81, ebioscience) at a final concentration of 5 ug/ml and PHA (Phytohaemagglutinin) (L8754-50, sigma) at a final concentration of 5 ug/ml respectively, incubating for 12 hrs at 37° C in 5% CO2. .. With BD Golgistop protein transport inhibitor (554715, BD) containing monensin at a final concentration of 6 μg/ml, incubating for an additional 5 hrs at 37° C in 5% CO2. (4) After a total of 17 hours cell culture preparation, mononuclear cells were washed and stained with cell surface NK/CD56+ T cell markers, CD56-Pacific Blue and CD3-PECy(7557851, BD), T cells markers, CD8-Alexa FLOUR 488(557696, BD), and CD4-PECy5 (555348, BD) for 30 min. .. Samples were then fixed and permeabilized according to the manufacturer’s instructions and stained for intracellular IFN-γ-APC (554702, BD) and IL-10-PE (16-7108-85, ebioscience) for an additional 30 min. (5) After washing, cells were re-suspended in 1% paraformaldehyde (AR1068, Boster Wuhan) until five-color flow cytometric analysis was performed on a LSRII instrument (BD Biosciences).

Modification:

Article Title: Friend retrovirus infection induces the development of memory-like natural killer cells
Article Snippet: Zombie UV Fixable Viability Kit (BioLegend) was used for the exclusion of dead cells. .. If stimulation was necessary, cells were stimulated with ionomycin (500 ng/ml), phorbol myristate acetate (PMA; 25 ng/ml), monensin (1X), and brefeldin A (2 µg/ml) diluted in Iscove’s modified Dulbecco’s medium (IMDM) buffer at 37 °C for 3 h. For intracellular stainings, cells were fixed with the Cytofix/Cytoperm Fixation/Permeabilization kit (BD) or with Foxp3/Transcription Factor staining buffer set (eBioscience). .. Cells were measured at LSR II (BD).

Concentration Assay:

Article Title: Description of organ-specific phenotype, and functional characteristics of tissue resident lymphocytes from liver transplantation donor and research on immune tolerance mechanism of liver
Article Snippet: Adding anti-human CD3(16-0037-85, e-bioscience) (clone:OKT3), at a final concentration of 10 ug/ml to 100 ul PBS(0014217, BI) solution, and putting on a packet plate (352360, BD Falcon) and incubating for 3 hours at 37° C to spare; (2) Washing the plate twice with 200 ul cold PBS solution; putting mononuclear cells from peripheral blood, lymph node, spleen, and liver perfusion on the plates (each plate contains 1 million cells). (3) Adding RPMI 1640 medium, anti-human CD28 (clone:CD28) (16-0289-81, ebioscience) at a final concentration of 5 ug/ml and PHA (Phytohaemagglutinin) (L8754-50, sigma) at a final concentration of 5 ug/ml respectively, incubating for 12 hrs at 37° C in 5% CO2. .. With BD Golgistop protein transport inhibitor (554715, BD) containing monensin at a final concentration of 6 μg/ml, incubating for an additional 5 hrs at 37° C in 5% CO2. (4) After a total of 17 hours cell culture preparation, mononuclear cells were washed and stained with cell surface NK/CD56+ T cell markers, CD56-Pacific Blue and CD3-PECy(7557851, BD), T cells markers, CD8-Alexa FLOUR 488(557696, BD), and CD4-PECy5 (555348, BD) for 30 min. .. Samples were then fixed and permeabilized according to the manufacturer’s instructions and stained for intracellular IFN-γ-APC (554702, BD) and IL-10-PE (16-7108-85, ebioscience) for an additional 30 min. (5) After washing, cells were re-suspended in 1% paraformaldehyde (AR1068, Boster Wuhan) until five-color flow cytometric analysis was performed on a LSRII instrument (BD Biosciences).

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    Becton Dickinson monensin
    Fine characterization of the immunodominant CD4 T cell response specific to HpaA142-159 in individual 47 ( A ) The 13mer overlapping peptides within the HpaA142-159 18mer were screened, with the 18mer results being shown as open bar. ( B ) Three neighboring 13mer peptides were titrated to compare their activities for identifying the most potent core sequence. HLA class II antibodies ( C ) and partial HLA class II matched BLCLs ( D and E ) were used to identify the HLA allele presenting the HpaA144-156 13mer peptide. ( F ) Partial HLA class II matched BLCLs were pulsed with recombinant HpaA for 24 hours and then co-cultured with HpaA-specific T cells for 5 hours in the presence of <t>monensin.</t> IFN-γ-producing CD4 + T cells were determined by ICS to determine whether the epitope HpaA144-156 was a naturally processed peptide.
    Monensin, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monensin/product/Becton Dickinson
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monensin - by Bioz Stars, 2021-03
    86/100 stars
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    Fine characterization of the immunodominant CD4 T cell response specific to HpaA142-159 in individual 47 ( A ) The 13mer overlapping peptides within the HpaA142-159 18mer were screened, with the 18mer results being shown as open bar. ( B ) Three neighboring 13mer peptides were titrated to compare their activities for identifying the most potent core sequence. HLA class II antibodies ( C ) and partial HLA class II matched BLCLs ( D and E ) were used to identify the HLA allele presenting the HpaA144-156 13mer peptide. ( F ) Partial HLA class II matched BLCLs were pulsed with recombinant HpaA for 24 hours and then co-cultured with HpaA-specific T cells for 5 hours in the presence of monensin. IFN-γ-producing CD4 + T cells were determined by ICS to determine whether the epitope HpaA144-156 was a naturally processed peptide.

    Journal: Oncotarget

    Article Title: Systematic identification of immunodominant CD4+ T cell responses to HpaA in Helicobacter pylori infected individuals

    doi: 10.18632/oncotarget.11092

    Figure Lengend Snippet: Fine characterization of the immunodominant CD4 T cell response specific to HpaA142-159 in individual 47 ( A ) The 13mer overlapping peptides within the HpaA142-159 18mer were screened, with the 18mer results being shown as open bar. ( B ) Three neighboring 13mer peptides were titrated to compare their activities for identifying the most potent core sequence. HLA class II antibodies ( C ) and partial HLA class II matched BLCLs ( D and E ) were used to identify the HLA allele presenting the HpaA144-156 13mer peptide. ( F ) Partial HLA class II matched BLCLs were pulsed with recombinant HpaA for 24 hours and then co-cultured with HpaA-specific T cells for 5 hours in the presence of monensin. IFN-γ-producing CD4 + T cells were determined by ICS to determine whether the epitope HpaA144-156 was a naturally processed peptide.

    Article Snippet: Intracellular cytokine staining (ICS) In the 18mer and 13mer peptide screening assays, bulk cultured T cells were incubated with peptide at 5μM in “RP-5” at 37°C for 5h in the presence of monensin (Becton Dickinson).

    Techniques: Sequencing, Recombinant, Cell Culture

    NK cells from Rae-1 Tg and WT mice produce comparable amounts of IFN-γ and CD107a when stimulated with Ly49H agonists. A and B , Freshly isolated splenocytes from Rae-1 Tg or WT littermate controls were enriched for NK cells and stimulated with plate-bound Abs against the NKRs NKG2D, Ly49D, and Ly49H or an isotype-matched control Ig in the presence of monensin (BD GolgiStop). After 4 h, cells were stained for intracellular IFN-γ and surface CD107a expression. Plots show the percentage of NK1.1 + TCRβ − cells that produce ( A ) IFN-γ and ( B ) CD107a. C and D , NK cells enriched as in A were incubated with Ba/F3 or Ba/F3-m157 targets in the presence of monensin. After 4 h, cells were stained for intracellular IFN-γ and surface CD107a expression. Plots show the percentage of NK1.1 + TCRβ − cells that produce ( C ) IFN-γ and ( D ) CD107a. Data are representative of three experiments.

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

    Article Title: Intact NKG2D-Independent Function of NK Cells Chronically Stimulated with the NKG2D Ligand Rae-1

    doi: 10.4049/jimmunol.1000397

    Figure Lengend Snippet: NK cells from Rae-1 Tg and WT mice produce comparable amounts of IFN-γ and CD107a when stimulated with Ly49H agonists. A and B , Freshly isolated splenocytes from Rae-1 Tg or WT littermate controls were enriched for NK cells and stimulated with plate-bound Abs against the NKRs NKG2D, Ly49D, and Ly49H or an isotype-matched control Ig in the presence of monensin (BD GolgiStop). After 4 h, cells were stained for intracellular IFN-γ and surface CD107a expression. Plots show the percentage of NK1.1 + TCRβ − cells that produce ( A ) IFN-γ and ( B ) CD107a. C and D , NK cells enriched as in A were incubated with Ba/F3 or Ba/F3-m157 targets in the presence of monensin. After 4 h, cells were stained for intracellular IFN-γ and surface CD107a expression. Plots show the percentage of NK1.1 + TCRβ − cells that produce ( C ) IFN-γ and ( D ) CD107a. Data are representative of three experiments.

    Article Snippet: After 4 h at 37°C in the presence of monensin (BD GolgiStop), cells were washed and stained for intracellular IFN-γ by using an Intracellular Staining kit ( ).

    Techniques: Mouse Assay, Isolation, Staining, Expressing, Incubation

    Increased production of IL-17 cytokine in the latent tuberculosis infection controls compared with patients with TB infection . Peripheral blood mononuclear cells from subjects with latent tuberculosis infection (n=26) and patients with TB infection (n=28) were stimulated with 50 ng/ml PMA, 1 μg/ml Ionomycin, 3 μM monensin for 4–5 hr. frequency of IL-17 producing CD4 + T cells was measured by intracellular staining and flowcytometry. Results showing that production of IL-17 cytokine was significantly increased in the (a) latent TB and (b) patients with active TB ( P =0.003)

    Journal: Iranian Journal of Basic Medical Sciences

    Article Title: Evaluation of Interleukin17and Interleukin 23 expression in patients with active and latent tuberculosis infection

    doi:

    Figure Lengend Snippet: Increased production of IL-17 cytokine in the latent tuberculosis infection controls compared with patients with TB infection . Peripheral blood mononuclear cells from subjects with latent tuberculosis infection (n=26) and patients with TB infection (n=28) were stimulated with 50 ng/ml PMA, 1 μg/ml Ionomycin, 3 μM monensin for 4–5 hr. frequency of IL-17 producing CD4 + T cells was measured by intracellular staining and flowcytometry. Results showing that production of IL-17 cytokine was significantly increased in the (a) latent TB and (b) patients with active TB ( P =0.003)

    Article Snippet: Intracellular cytokine staining and flowcytometric analysis To measure intracellular IL-17, PBMCs after culturing with PPD were stimulated with PMA (50 ng/ml) plus ionomycin (1 μg/ml) and monensin (3 μM Monensin) (BD, USA) for 4 hr in 1 ml RPMI 1640 media containing 10% fetal calf serum (FCS).

    Techniques: Infection, Staining

    The natural processing and presentation of L 57–69 and L 83–95 by DCs. (A) The DCs were pulsed with L 57–69 , rLpp20, HP-WCL, or BSA for 24 h and then co-cultured with L 57–69 -specific T cells from subject H6 for 5 h in the presence of monensin. The frequency of IFN-γ-secreting CD4 + T cells was determined by ICS. (B) L 83–95 -specific T cells from subject H1 were used to determine in a manner similar to the peptide described in (A) .

    Journal: Frontiers in Microbiology

    Article Title: Identification of Two Lpp20 CD4+ T Cell Epitopes in Helicobacter pylori-Infected Subjects

    doi: 10.3389/fmicb.2018.00884

    Figure Lengend Snippet: The natural processing and presentation of L 57–69 and L 83–95 by DCs. (A) The DCs were pulsed with L 57–69 , rLpp20, HP-WCL, or BSA for 24 h and then co-cultured with L 57–69 -specific T cells from subject H6 for 5 h in the presence of monensin. The frequency of IFN-γ-secreting CD4 + T cells was determined by ICS. (B) L 83–95 -specific T cells from subject H1 were used to determine in a manner similar to the peptide described in (A) .

    Article Snippet: Intracellular cytokine staining (ICS) In the assays for screening immunodominant CD4+ T cell epitopes by 18mer and 13mer synthetic peptide, bulk cultured Lpp20-specific T cells were incubated with 5 μmol/L peptide at 37°C for 5 h in the presence of monensin (Becton Dickinson, Shanghai, China).

    Techniques: Cell Culture