High avidity antigen-specific CTL identified by CD8-independent tetramer staining.
Tetrameric MHC/peptide complexes are important tools for enumerating, phenotyping, and rapidly cloning Ag-specific T cells. It remains however unclear whether they can reliably distinguish between high and low avidity T cell clones. In this report, tetramers with mutated CD8 binding site selectively stain higher avidity human and murine CTL capable of recognizing physiological levels of Ag. Furthermore, we demonstrate that CD8 binding significantly enhances the avidity as well as the stability of interactions between CTL and cognate tetramers. The use of CD8-null tetramers to identify high avidity CTL provides a tool to compare vaccination strategies for their ability to enhance the frequency of high avidity CTL. Using this technique, we show that DNA priming and vaccinia boosting of HHD A2 transgenic mice fail to selectively expand large numbers of high avidity NY-ESO-1(157-165)-specific CTL, possibly due to the large amounts of antigenic peptide delivered by the vaccinia virus. Furthermore, development of a protocol for rapid identification of high avidity human and murine T cells using tetramers with impaired CD8 binding provides an opportunity not only to monitor expansion of high avidity T cell responses ex vivo, but also to sort high avidity CTL clones for adoptive T cell transfer therapy.
Choi
Ed Man-Lik
EM
Tumour Immunology Unit, Nuffield Department of Medicine, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, Oxford, United Kingdom.
Chen
Ji-Li
JL
Wooldridge
Linda
L
Salio
Mariolina
M
Lissina
Anna
A
Lissin
Nikolai
N
Hermans
Ian F
IF
Silk
Jonathan D
JD
Mirza
Fareed
F
Palmowski
Michael J
MJ
Dunbar
P Rod
PR
Jakobsen
Bent K
BK
Sewell
Andy K
AK
Cerundolo
Vincenzo
V
D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics / Q000276:immunology; D016827:CD8 Antigens / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D002999:Clone Cells; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D006183:H-2 Antigens / Q000235:genetics / Q000378:metabolism; D015789:HLA-A2 Antigen / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007117:Immunization, Secondary; D019169:Jurkat Cells; D008213:Lymphocyte Activation / Q000235:genetics; D008565:Membrane Proteins; D051379:Mice; D008822:Mice, Transgenic; D010957:Plasmids / Q000008:administration & dosage; D011506:Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling; D013602:T-Lymphocytes, Cytotoxic / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D014615:Vaccinia / Q000235:genetics / Q000276:immunology; D001613:beta 2-Microglobulin / Q000032:analysis / Q000378:metabolism
D000951:Antigens, Neoplasm; D016827:CD8 Antigens; C104948:CTAG1B protein, human; D018984:Epitopes, T-Lymphocyte; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D015789:HLA-A2 Antigen; D008565:Membrane Proteins; D011506:Proteins; D001613:beta 2-Microglobulin