Use of class II tetramers for identification of CD4+ T cells.
Multivalent MHC class II molecules containing peptide antigens are useful tools for the detection of antigen specific human CD4+ T cells. Tetramers produced by exogenous peptide loading onto empty class II molecules are comparable to tetramers with peptide tethered to the class II chain covalently, but have many practical advantages. Conditions for optimal peptide loading to generate tetramers are discussed and optimal conditions of using tetramers for staining T cells are examined. As the frequency of antigen specific CD4+ T cells in peripheral blood is low, we demonstrate that an in vitro expansion step is effective in detecting low frequency T cells. Two new applications with tetramers, their uses for mapping T cell epitopes and for the detection of low affinity T cells are described. In a clinical setting, potential applications include using these reagents for monitoring disease progression during clinical intervention.
Kwok
William W
WW
The Benaroya Research Institute at Virginia Mason, 1201 Ninth Avenue, Seattle, WA 98101, USA. bkwok@vmresearch.org
Ptacek
Nancy A
NA
Liu
Andrew W
AW
Buckner
Jane H
JH
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D019295:Computational Biology; D018604:Epitope Mapping; D006683:HLA-DQ Antigens / Q000737:chemistry / Q000378:metabolism; D006684:HLA-DR Antigens / Q000737:chemistry / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D013194:Staining and Labeling; D013696:Temperature
D006683:HLA-DQ Antigens; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II