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I ab gp66 77 specific cd4 t cells

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I-Ab/GP66–77-specific CD4 T cells are a type of T lymphocyte that recognizes and binds to the I-Ab major histocompatibility complex (MHC) molecule presenting the GP66–77 peptide. They are used in research applications to study T cell responses and immune function.

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Lab products found in correlation

Fixation permeabilization kit, by BD (2 mentions) Cd44 clone im7, by BioLegend (2 mentions) Thy1.1 clone ox 7, by BioLegend (2 mentions) Ifnγ clone xmg1, by BD (2 mentions) Kavynfa, by AnaSpec (2 mentions) Thy1.2 clone 30 h12, by BioLegend (2 mentions) Cd8 clone 53 6, by BioLegend (2 mentions) Tnfα clone mp6 xt22, by Thermo Fisher Scientific (2 mentions) Cd4 clone gk1, by BioLegend (2 mentions) Lsrii flow cytometer, by BD (2 mentions)

2 protocols using i ab gp66 77 specific cd4 t cells

1

Flow Cytometric Analysis of Antigen-Specific T Cells

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For antigen-specific restimulation, 1 × 106 splenocytes were cultured for 6 hrs at 37°C in 5% CO2 in complete RPMI-1640 (10% HI-FBS, 55 μM beta-mercaptoethanol, 1x Pen/Strep and 1x L-Glut) supplemented with LCMV GP33–41 peptide KAVYNFATM (0.5 μg/mL, AnaSpec, cat. AS-61296) or left unstimulated.
For flow cytometric analysis, 1 × 106 splenocytes or LN cells were stained with antibodies specific for surface markers Thy1.2 (clone 30-H12, BioLegend), Thy1.1 (clone OX-7, BioLegend), CD8 (clone 53–6.7, BioLegend), CD4 (clone GK1.5, BioLegend), CD44 (clone IM7, BioLegend) and tetramers for either H2Db/GP33–43-specific CD8 T cells, H2Db/NP396–404-specific CD8 T cells or I-Ab/GP66–77-specific CD4 T cells (all from NIH Tetramer Core Facility). For intracellular staining, cells were processed using the Fixation/Permeabilization kit from BD Biosciences (BD Cytofix/Cytoperm, cat. 554714) following manufacturer’s instructions and stained with antibodies specific for IFNγ (clone XMG1.2, BD Biosciences) and TNFα (clone MP6-XT22, eBioscience). Samples were analysed on a BD Biosciences LSRII flow cytometer. Analysis of frequency and mean fluorescence intensity (MFI) was performed using FlowJo software. MFI was defined as the geometric mean of the fluorescence signal of interest for a given gated population.
Damo M., Fitzgerald B., Lu Y., Nader M., William I., Cheung J.F., Connolly K.A., Foster G.G., Akama-Garren E., Lee D.Y., Chang G.P., Gocheva V., Schmidt L.M., Boileve A., Wilson J.H., Cui C., Monroy I., Gokare P., Cabeceiras P., Jacks T, & Joshi N.S. (2020). Inducible de novo expression of neoantigens in tumor cells and mice with NINJA. Nature biotechnology, 39(1), 64-73.
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2

Flow Cytometric Analysis of Antigen-Specific T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
For antigen-specific restimulation, 1 × 106 splenocytes were cultured for 6 hrs at 37°C in 5% CO2 in complete RPMI-1640 (10% HI-FBS, 55 μM beta-mercaptoethanol, 1x Pen/Strep and 1x L-Glut) supplemented with LCMV GP33–41 peptide KAVYNFATM (0.5 μg/mL, AnaSpec, cat. AS-61296) or left unstimulated.
For flow cytometric analysis, 1 × 106 splenocytes or LN cells were stained with antibodies specific for surface markers Thy1.2 (clone 30-H12, BioLegend), Thy1.1 (clone OX-7, BioLegend), CD8 (clone 53–6.7, BioLegend), CD4 (clone GK1.5, BioLegend), CD44 (clone IM7, BioLegend) and tetramers for either H2Db/GP33–43-specific CD8 T cells, H2Db/NP396–404-specific CD8 T cells or I-Ab/GP66–77-specific CD4 T cells (all from NIH Tetramer Core Facility). For intracellular staining, cells were processed using the Fixation/Permeabilization kit from BD Biosciences (BD Cytofix/Cytoperm, cat. 554714) following manufacturer’s instructions and stained with antibodies specific for IFNγ (clone XMG1.2, BD Biosciences) and TNFα (clone MP6-XT22, eBioscience). Samples were analysed on a BD Biosciences LSRII flow cytometer. Analysis of frequency and mean fluorescence intensity (MFI) was performed using FlowJo software. MFI was defined as the geometric mean of the fluorescence signal of interest for a given gated population.
Damo M., Fitzgerald B., Lu Y., Nader M., William I., Cheung J.F., Connolly K.A., Foster G.G., Akama-Garren E., Lee D.Y., Chang G.P., Gocheva V., Schmidt L.M., Boileve A., Wilson J.H., Cui C., Monroy I., Gokare P., Cabeceiras P., Jacks T, & Joshi N.S. (2020). Inducible de novo expression of neoantigens in tumor cells and mice with NINJA. Nature biotechnology, 39(1), 64-73.
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