title: Cutting edge: detection of antigen-specific CD4+ T cells by HLA-DR1 oligomers is dependent on the T cell activation state.
abstract: Class I MHC tetramers have proven to be invaluable tools for following and deciphering the CD8(+) T cell response, but the development of similar reagents for detection of CD4(+) T cells based on class II MHC proteins has been more difficult. We evaluated fluorescent streptavidin-based oligomers of HLA-DR1 for use as reagents to analyze Ag-specific human CD4(+) T cells. Staining was blocked at low temperatures and by drugs that disrupt microfilament formation and endocytosis. Cell-associated MHC oligomers were resistant to a surface stripping protocol and were observed by microscopy in intracellular compartments. This behavior indicates that detection of CD4(+) T cells using class II MHC oligomers can depend on an active cellular process in which T cells cluster and/or endocytose their Ag receptors. T cells of identical specificity but in different activation states varied greatly in their ability to be detected by class II MHC oligomers.
authors.affiliation: Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: T O
authors.initials: TO
authors.lastname: Cameron
authors.forename: J R
authors.initials: JR
authors.lastname: Cochran
authors.forename: B
authors.initials: B
authors.lastname: Yassine-Diab
authors.forename: R P
authors.initials: RP
authors.lastname: Sékaly
authors.forename: L J
authors.initials: LJ
authors.lastname: Stern
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000032:analysis; D019921:Biotinylation; D015496:CD4-Positive T-Lymphocytes / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D002451:Cell Compartmentation / Q000276:immunology; D002460:Cell Line; D017635:Clonal Anergy; D002999:Clone Cells; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000378:metabolism; D015798:HLA-DR1 Antigen / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000276:immunology; D013194:Staining and Labeling / Q000379:methods; D016176:T-Lymphocyte Subsets / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D018984:Epitopes, T-Lymphocyte; D015798:HLA-DR1 Antigen