Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers.
The use of fluorescently labeled major histocompatibility complex multimers has become an essential technique for analyzing disease- and therapy-induced T-cell immunity. Whereas classical major histocompatibility complex multimer analyses are well-suited for the detection of immune responses to a few epitopes, limitations on human-subject sample size preclude a comprehensive analysis of T-cell immunity. To address this issue, we developed a combinatorial encoding strategy that allows the parallel detection of a multitude of different T-cell populations in a single sample. Detection of T cells from peripheral blood by combinatorial encoding is as efficient as detection with conventionally labeled multimers but results in a substantially increased sensitivity and, most notably, allows comprehensive screens to be performed. We obtained proof of principle for the feasibility of large-scale screening of human material by analysis of human leukocyte antigen A3-restricted T-cell responses to known and potential melanoma-associated antigens in peripheral blood from individuals with melanoma.
Hadrup
Sine Reker
SR
Division of Immunology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Bakker
Arnold H
AH
Shu
Chengyi J
CJ
Andersen
Rikke S
RS
van Veluw
Jerre
J
Hombrink
Pleun
P
Castermans
Emilie
E
Thor Straten
Per
P
Blank
Christian
C
Haanen
John B
JB
Heemskerk
Mirjam H
MH
Schumacher
Ton N
TN
D000941:Antigens; D000951:Antigens, Neoplasm; D002469:Cell Separation / Q000379:methods; D000939:Epitopes; D005456:Fluorescent Dyes; D006649:Histocompatibility Antigens / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007158:Immunologic Techniques; D058950:Melanoma-Specific Antigens; D036103:Nanotechnology; D009363:Neoplasm Proteins; D010455:Peptides / Q000276:immunology; D020836:Protein Structure, Quaternary; D045663:Quantum Dots; D012680:Sensitivity and Specificity; D016176:T-Lymphocyte Subsets / Q000145:classification / Q000166:cytology / Q000276:immunology*
D000941:Antigens; D000951:Antigens, Neoplasm; D000939:Epitopes; D005456:Fluorescent Dyes; D006649:Histocompatibility Antigens; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; D010455:Peptides