"Exon-shuffling" maps control of antibody- and T-cell-recognition sites to the NH2-terminal domain of the class II major histocompatibility polypeptide A beta.
"Exon-shuffling" maps control of antibody- and T-cell-recognition sites to the NH2-terminal domain of the class II major histocompatibility polypeptide A beta.
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“外显子改组”将抗体和 T 细胞识别位点的控制映射到 II 类主要组织相容性多肽 Aβ 的 NH2 末端结构域。
DOI：
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发表时间：
1985
影响因子：
11.1
通讯作者：
J. Tou
中科院分区：
文献类型：
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作者：
R. Germain;J. Ashwell;R. Lechler;D. Margulies;K. M. Nickerson;G. Suzuki;J. Tou
为了研究第二类主要组织相容性多肽Aβ的高度多态的氨基末端(β1)结构域在T细胞和抗体识别中的作用，在AKβ和Adβ的基因组重组DNA克隆之间进行了“外显子改组”，以产生一个包含AKβ氨基末端结构域的AKβ外显子的杂交基因，然后是分子其余部分的Adβ外显子。将此杂合Aβ基因与AKα联合表达的L细胞基因转染体与表达野生型AKβAKα二聚体的L细胞进行T细胞克隆和杂交瘤的抗原提呈试验，并用一组抗I-AK特异性单抗进行染色。这些抗体还被测试了它们与在没有AKα的情况下表达AKβ的B淋巴瘤转染体的反应性。结果表明，无论是T细胞还是抗体介导的对含有外显子改组或野生型AKβ的I-AK分子的识别没有质的差异。结合只表达AKβ的B细胞转染者的数据，这些结果将控制Aβ对免疫相关的I-AK决定因素的贡献映射到高度多态的氨基末端区域，并表明Aβ多肽其余部分的氨基酸序列变化对等位基因特异性识别的贡献很小(如果有的话)。
To investigate the role of the highly polymorphic amino-terminal (beta 1) domain of the class II major histocompatibility polypeptide A beta during recognition by T cells and antibodies, "exon-shuffling" was carried out between genomic recombinant DNA clones of Ak beta and Ad beta to generate a hybrid gene containing Ak beta exons for the amino-terminal domain followed by the Ad beta exons for the remainder of the molecule. L-cell gene transfectants expressing this hybrid A beta gene in combination with Ak alpha were compared to L cells expressing wild-type Ak beta Ak alpha dimers in tests of antigen-presentation to T-cell clones and hybridomas and for staining by a panel of anti-I-Ak-specific monoclonal antibodies. These antibodies were also tested for their reactivity with a B-lymphoma transfectant expressing Ak beta in the absence of Ak alpha. The results showed no qualitative differences in either T-cell or antibody-mediated recognition of I-Ak molecules containing either the exon-shuffled or wildtype Ak beta. Together with the data involving the B cell transfectant expressing only Ak beta, these results map control of the A beta contribution to the immunologically relevant determinants of I-Ak to the highly polymorphic amino-terminal domain and indicate little, if any, contribution to allele-specific recognition by amino acid sequence variations in the remaining portions of the A beta polypeptide.