A very rapid and simple assay based on trogocytosis to detect and measure specific T and B cell reactivity by flow cytometry.
Detection, quantification, separation and characterization of T and B cells reactive to specific antigens are important tasks in both basic and clinical immunology. Here, we describe an approach allowing the performance of all four tasks on a functional basis by flow cytometry. The assay is based on the property of lymphocytes to capture membrane components from the cells they interact with, in a process we call trogocytosis. Working with CD8+ CTL and target cells labeled with membrane markers, we describe the conditions allowing reactive lymphocytes to be detected rapidly and inexpensively within mixed populations. Accordingly, we used this method to monitor the CTL response triggered in mice after vaccination. In addition, we documented the applicability of this method to the detection of antigen-specific CD4+ T and B cells. While our method is, for the time being, not as sensitive as staining of CTL with MHC class I multimers, it allows the simultaneous quantitative identification of reactive CD8+, CD4+ and B cells. Altogether, our method offers a simple and general alternative to other methods previously described to detect and quantify lymphocyte reactivity, and it can also be used in combination with those.
Puaux
Anne-Laure
AL
CPTP, INSERM U563, Institut Claude de Préval, Toulouse, France.
Campanaud
Julie
J
Salles
Audrey
A
Préville
Xavier
X
Timmerman
Benedikt
B
Joly
Etienne
E
Hudrisier
Denis
D
D000818:Animals; D030801:Animals, Genetically Modified; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002450:Cell Communication / Q000276:immunology*; D045744:Cell Line, Tumor; D002469:Cell Separation; D005434:Flow Cytometry / Q000379:methods*; D007111:Immunity, Cellular / Q000276:immunology*; D051379:Mice; D012680:Sensitivity and Specificity; D014611:Vaccination / Q000379:methods