T cell enrichment by cytotoxic elimination of B cells and accessory cells.
T cells from mouse spleen and lymph node do not express the cell-surface glycoproteins encoded for by MHC class II genes, whereas most non-T cells in these organs do (i.e., B cells and accessory cells). This unique feature of T cells makes it possible to enrich for them using cytotoxic anti-class II monoclonal antibodies and activating complement. Because there are two isotypes of MHC class II genes (A and E genes, located adjacent to each other in the MHC region originally called the I region), it is necessary to identify which of these two genes are expressed in the mouse strain used. This unit lists suitable antibody-producing hybridomas available from ATCC, with antigen specificities (I-A vs. I-E) and corresponding mouse strains (H-2 strains a, b, k, etc., indicated as superscripts). Other antibodies can be tested in pilot experiments using the procedure outlined. This unit describes T cell enrichment using cytotoxic antibodies, and UNIT 3.4 describes the depletion of T cells and their subpopulations using the same approach. In the latter unit, T cell surface markers (Thy-1, CD4, and CD8) are targeted by the cytotoxic antibodies.
Hathcock
K
K
National Cancer Institute, Bethesda, Maryland, USA.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology; D015704:CD4 Antigens / Q000276:immunology; D016827:CD8 Antigens / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D003602:Cytotoxicity, Immunologic; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D008198:Lymph Nodes / Q000276:immunology; D051379:Mice; D013154:Spleen / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D018800:Thy-1 Antigens / Q000276:immunology
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D016827:CD8 Antigens; D000949:Histocompatibility Antigens Class II; D018800:Thy-1 Antigens