Charmley P, Nickerson D, Hood L
Virginia Mason Research Center, Seattle, WA 98101.
Immunogenetics. 1994;39(2):138-45. doi: 10.1007/BF00188617.
The T-cell receptor (Tcr) provides specificity for antigen recognition by its variable domain, primarily consisting of two germline encoded variable (V) region gene segments. Thus it has been suggested that inherited polymorphisms in the TCRV gene segments could contribute to differential immune responsiveness (e.g., autoimmunity) in human populations. In the present study, we have sought potentially functional polymorphisms in the germline TCRAV gene segments. Using denaturing gradient gel electrophoresis on polymerase chain reaction (PCR)-amplified products from the pooled DNA of many individuals, we identified polymorphisms in the TCRAV2S1, AV4S1, AV7S1, and AV8S1 gene segments. A complete DNA sequence analysis of these PCR products identified polymorphisms that affected amino acids in the predicted antigen-binding regions of the Tcr alpha chain, as well as polymorphisms in the introns. Genotype analysis of all nine DNA point mutations showed a 5%-50% range (averaging 35%) of minor allele frequencies, often resulting in individuals homozygous for the alternate allele forms. All possible haplotype combinations of the amino acid-affecting polymorphisms were found, indicating that in human populations there are a large number of different germline haplotypes encoding V gene segment alleles. These TCRAV coding region polymorphisms provide the rationale for, and allow the direct testing of, hypotheses concerning inherited polymorphisms within the T-cell receptor genes that may contribute to autoimmune susceptibility.
T细胞受体(Tcr)通过其可变结构域提供抗原识别特异性,该可变结构域主要由两个种系编码的可变(V)区基因片段组成。因此,有人提出TCRV基因片段中的遗传多态性可能导致人类群体中免疫反应性的差异(例如自身免疫)。在本研究中,我们寻找种系TCRAV基因片段中潜在的功能性多态性。通过对来自许多个体混合DNA的聚合酶链反应(PCR)扩增产物进行变性梯度凝胶电泳,我们在TCRAV2S1、AV4S1、AV7S1和AV8S1基因片段中鉴定出多态性。对这些PCR产物进行完整的DNA序列分析,确定了影响Tcrα链预测抗原结合区域氨基酸的多态性以及内含子中的多态性。对所有九个DNA点突变的基因型分析显示,次要等位基因频率范围为5%-50%(平均35%),常常导致个体为替代等位基因形式的纯合子。发现了所有影响氨基酸的多态性的可能单倍型组合,这表明在人类群体中有大量不同的种系单倍型编码V基因片段等位基因。这些TCRAV编码区多态性为关于T细胞受体基因内可能导致自身免疫易感性的遗传多态性的假说提供了理论依据,并允许对这些假说进行直接检验。