Cell-free translation of a biologically active, antigen-specific suppressor T cell factor.
Cell-free translation of a biologically active, antigen-specific suppressor T cell factor.
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具有生物活性、抗原特异性抑制性 T 细胞因子的无细胞翻译。
DOI：
10.1073/pnas.79.11.3599
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发表时间：
1982
影响因子：
11.1
通讯作者：
Webb,DR
中科院分区：
文献类型：
--
作者：
Wieder,KJ;Araneo,BA;Kapp,JA;Webb,DR
在兔网织红细胞裂解物无细胞翻译系统中，用部分纯化的含聚(A)的RNA体外合成了抗原特异性T细胞抑制因子(TSF)。从克隆的T细胞杂交瘤中分离到含PolyA的mRNA，该杂交瘤可产生合成多肽抗原Poly-(LGlu60LAla30LTyr10)(GAT)的特异性TSF。RNA按大小分级，并在体外翻译。16S RNA组分刺激一种生物活性蛋白的合成，该蛋白在体外特异性地抑制脾细胞对GAT的特异性抗体反应，以及GAT诱导的小鼠淋巴T细胞对GAT的增殖反应。此外，抑制因子与GAT结合，GAT是主要组织相容性复合体(MHC)I亚区编码的决定簇，通过NadodSO4/聚丙烯酰胺凝胶电泳分离的蛋白质的功能分析估计其表观MR为19,000。这些结果表明，获得具有生物活性的TSF几乎不需要翻译后修饰(除了蛋白水解性切割)。因此，碳水化合物或其他化学基团的存在既不影响GAT-TSF的血清学性质，也不影响其生物学性质。
In vitro synthesis of an antigen-specific T cell suppressor factor (TsF) has been accomplished by using partially purified poly(A)-containing RNA in a rabbit reticulocyte lysate cell-free translation system. The poly(A)-containing mRNA was isolated from a cloned T cell hybridoma that constitutively produces a TsF specific for the synthetic polypeptide antigen poly-(LGlu60LAla30LTyr10) (GAT). The RNA was fractionated by size and translated in vitro. The 16S RNA fraction stimulated synthesis of a biologically active protein that specifically suppressed both the GAT-specific antibody response by spleen cells in vitro and the proliferation response to GAT by lymph node T cells from GAT-primed mice. Further, the suppressor factor had a binding site for GAT, a determinant encoded by the I subregion of the major histocompatibility complex (MHC), and an apparent Mr 19,000 estimated by functional assays on protein separated by NadodSO4/polyacrylamide gel electrophoresis. These results indicate that virtually no posttranslational modifications (other than proteolytic cleavage) are necessary to obtain biologically active TsF. Hence, the presence of carbohydrate or other chemical groups does not contribute to either the serological properties of GAT-TsF or its biological properties.