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Cutting Edge: Detection of Antigen-Specific CD4⁺ T Cells by HLA-DR1 Oligomers Is Dependent on the T Cell Activation State¹

Thomas O. Cameron^*, Jennifer R. Cochran^*, Bader Yassine-Diab, Rafick-Pierre Sékaly and Lawrence J. Stern^2,*

^* Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139; and Centre de Recherche du Centre Hospitalier de l’Université de Montréal, Montréal, Canada

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.

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