Design of T-cell receptor libraries with diverse binding properties to examine adoptive T-cell responses.
Adoptive T-cell therapies have shown significant promise in the treatment of cancer and viral diseases. One approach, which introduces antigen-specific T-cell receptors (TCRs) into ex vivo activated T cells, is designed to overcome central tolerance mechanisms that prevent responses by endogenous T-cell repertoires. Studies have suggested that use of higher-affinity TCRs against class I major histocompatibility complex antigens could drive the activity of both CD4(+) and CD8(+) T cells, but the rules that govern the TCR binding optimal for in vivo activity are unknown. Here, we describe a high-throughput platform of 'reverse biochemistry' whereby a library of TCRs with a wide range of binding properties to the same antigen is introduced into T cells and adoptively transferred into mice with antigen-positive tumors. Extraction of RNA from tumor-infiltrating lymphocytes (TILs) or lymphoid organs allowed high-throughput sequencing to determine which TCRs were selected in vivo. The results showed that CD8(+) T cells expressing the highest-affinity TCR variants were deleted in both the TIL population and in peripheral lymphoid tissues. In contrast, these same high-affinity TCR variants were preferentially expressed within CD4(+) T cells in the tumor, suggesting they had a role in antigen-specific tumor control. The findings thus revealed that the affinity of the transduced TCRs controlled the survival and tumor infiltration of the transferred T cells. Accordingly, the TCR library strategy enables rapid assessment of TCR-binding properties that promote peripheral T-cell survival and tumor elimination.
Department of Biochemistry, University of Illinois, Urbana, IL 61801, USA.
A S
AS
Chervin
J D
JD
Stone
C M
CM
Soto
B
B
Engels
H
H
Schreiber
E J
EJ
Roy
D M
DM
Kranz
D056704:Adaptive Immunity / Q000235:genetics*; D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D064987:Cell- and Tissue-Based Therapy; D015723:Gene Library; D005822:Genetic Vectors; D015395:Histocompatibility Antigens Class I / Q000235:genetics; D006801:Humans; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology; D051379:Mice; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic
D015395:Histocompatibility Antigens Class I; D011948:Receptors, Antigen, T-Cell