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Immunology and Infection

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JoVE Journal Immunology and Infection
Quantification of Proliferating Human Antigen-specific CD4+ T Cells using Carboxyfluorescein Succinimidyl Ester
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Quantification of Proliferating Human Antigen-specific CD4+ T Cells using Carboxyfluorescein Succinimidyl Ester

Article DOI: 10.3791/59545-v • 07:00 min • June 4th, 2019
June 4th, 2019 •
1, 1, 1, 1,2
1Immunology and Diabetes Unit, St. Vincent's Institute of Medical Research, 2Department of Medicine, University of Melbourne, St. Vincent's Hospital

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Summary
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Presented here is a protocol for measuring proliferating CD4+ T cells in response to antigenic proteins or peptides using dye dilution. This assay is particularly sensitive to rare antigen-specific T cells and can be modified to facilitate cloning of antigen-specific cells.

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Quantification Proliferating Human Antigen-specific CD4+ T Cells Carboxyfluorescein Succinimidyl Ester Protocol Studying Weak Difficult To Detect Detection Enumeration Isolation Antigen Presentation Advantage Analysis Rare Populations Autoimmune Conditions Type One Diabetes Peripheral Blood Sample Dilute PBS Density Gradient Medium 50 Milliliter Tube Centrifugation Plasma Layer White Blood Cell Layer Red Blood Cell Pellet Wash Cells Fresh PBS Counting Cells Peripheral Blood Mononuclear Cell (PBMC) Concentration Control Tubes RP5 Medium Concentration
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