Use of immobilized HLA-A2:Ig dimeric proteins to determine the level of epitope-specific, HLA-restricted CD8(+) T-cell response.
A novel assay to assess antigen-specific cytokine release from stimulated CD8(+) T cells derived from the mucosal and peripheral blood compartments has been developed and standardized using the influenza A virus matrix protein (MP) peptide, GILGFVFTL. This technology is based on the capacity for the human leucocyte antigen (HLA)-A2:Ig dimeric protein to stimulate CD8(+) T cells in a major histocompatibility complex (MHC) class I-restricted fashion without the necessity for antigen presenting cells (APC). This assay has been optimized utilizing a 9-amino acid residue (9mer) peptide, the optimal peptide length for presenting an epitope to CD8(+) T cells. Compared to existing assays, this more sensitive and specific methodology requires fewer cells, enabling easier and more accurate monitoring of the CD8(+) T-cell response in biological compartments, such as the mucosa during the course of viral infection and may be utilized to assess epitope-specific CD8(+) T-cell responses in vaccine trials.
Horowitz
A
A
HIV and Malaria Vaccine Program, Aaron Diamond AIDS Research Center, The Rockefeller University, New York, NY 10016, USA.
Li
X
X
Poles
M A
MA
Tsuji
M
M
D017951:Antigen Presentation / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D015789:HLA-A2 Antigen / Q000276:immunology*; D006801:Humans; D007158:Immunologic Techniques; D008213:Lymphocyte Activation / Q000276:immunology*
D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen