A strategy for uncovering germline variants altering anti-tumor CD8 T cell response  

作　　者：Vijay Kumar Ulaganathan Martina H.Vasileva 

机构地区：[1]Klinik fur Dermatologie&Allergology,Universitatsmedizin Gottingen,Gottingen 37075,Germany [2]Institut fuir Multiple Sklerose Forschung,Neuroimmunologie,Universitatsmedizin Gottingen,Gottingen 37075,Germany [3]University of Lorraine,NGERE Unit,Faculte de Medecine,9 Avenue de La Foret de Haye,Vandoeuvre-les-Nancy 54505,France

出　　处：《Journal of Genetics and Genomics》2023年第5期353-361,共9页遗传学报（英文版）

摘　　要：Among many factors known to alter the outcomes of T cell receptor(TCR)-induced proximal signaling,the role of human germline variants in dictating the individuality of the anti-tumor CD8 T cell response has remained challenging to address.Here,we describe a convenient strategy for molecular and functional characterization of phosphotyrosine-altering non-synonymous single nucleotide variations(pTyr-SNVs)that directly impact TCR-induced proximal phosphotyrosine motif-based signaling pathways.We devise an experimental co-cultivation set-up comprising a C57BL/6 mouse-derived metastatic melanoma cell line engineered to constitutively present ovalbumin(OVA)antigens and retrovirally engineered syngeneic major histocompatibility complex(MHC)Class I restricted OVA TCR-transgenic CD8 T cells(OT-I).Using the synthetic version of pTyr-SNV rs1178800678-G/T-encoding integrin alpha 4(ITGA4)p.S1027I variant as a prototype,we show that under identical TCR stimulation conditions,genetically determined membrane-proximal immunoreceptor tyrosin activation motif(ITAM)results in increased tyrosine phosphorylation of 70 kDa zeta-chain-associated protein(ZAP70)and the levels of cytotoxic effector molecule granzyme B(GZMB),which in turn result in enhanced cytotoxic activity against metastatic melanoma cell line.This strategy paves the way for rapid molecular and functional characterization of anti-tumor immune response-linked germline pTyr-SNVs so as to improve our understanding of the genetic basis of individual-to-individual differences in anti-tumor CD8 T cell response.

关 键 词：Non-synonymous single nucleotide variants(nsSNVs) Membrane-proximal phosphotyrosine signalling motifs Phosphotyrosine-altering non-synonymous single nucleotide variants(pTyr-SNVs) Immunoreceptor tyrosine activation motif(ITAM) CD8 T cells Melanoma B16-F10 

分 类 号：R730.51[医药卫生—肿瘤]