Monitoring antigen-specific T cells using MHC-Ig dimers.
The lack of high affinity reagents has made distinguishing T cells on the basis of antigen specificity difficult to accomplish. This unit provides protocols that utilize innovations in molecular design to permit construction of soluble multivalent MHC complexes (MHC-Ig dimers) with high avidity for cognate T cell receptors. MHC-Ig dimers display stable binding properties when they interact with antigen-specific T cells thus allowing their use in the staining of antigen-specific T cells by flow cytometry. Methods for constructing and detecting these MHC-Ig dimers are included along with protocols for applying their use for the quantitation of antigen-specific T cells.
Schneck
J P
JP
Johns Hopkins University, Baltimore, Maryland, USA.
Slansky
J E
JE
O'Herrin
S M
SM
Greten
T F
TF
D000818:Animals; D000941:Antigens / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000276:immunology; D019281:Dimerization; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology
D000941:Antigens; D000954:Antigens, Surface; D018076:DNA, Complementary; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D007136:Immunoglobulins; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins