Kwon B S, Kim G S, Prystowsky M B, Lancki D W, Sabath D E, Pan J L, Weissman S M
Proc Natl Acad Sci U S A. 1987 May;84(9):2896-900. doi: 10.1073/pnas.84.9.2896.
A modified differential screening procedure was applied to analyze cDNA libraries of cloned helper T lymphocytes (Th) and cytolytic T lymphocytes (CTL). Negative and positive differential screening and RNA blot analysis were used to identify cDNA clones that were expressed preferentially in Th or CTL. Seven clones corresponded to previously described T-cell genes, and 16 additional types of cDNA clones were isolated, 9 from Th and 7 from CTL. Of these, 3 were expressed in both Th and CTL, 7 were expressed in only Th, and 6 only in CTL. These clones were analyzed for induction after stimulation by interleukin 2 or Con A or after stimulation of the T-cell antigen receptor (TCR). Three different patterns of expression were seen: induction only by Con A, induction by Con A and interleukin 2, and induction by Con A and TCR stimulation. The approach is potentially useful for analyzing paths of T-cell differentiation and detecting cDNA clones encoding unrecognized cytokines.
应用一种改良的差异筛选程序来分析克隆的辅助性T淋巴细胞（Th）和细胞毒性T淋巴细胞（CTL）的cDNA文库。采用阴性和阳性差异筛选以及RNA印迹分析来鉴定在Th或CTL中优先表达的cDNA克隆。7个克隆对应于先前描述的T细胞基因，另外分离出16种类型的cDNA克隆，9个来自Th，7个来自CTL。其中，3个在Th和CTL中均表达，7个仅在Th中表达，6个仅在CTL中表达。分析这些克隆在白细胞介素2或刀豆蛋白A刺激后或T细胞抗原受体（TCR）刺激后的诱导情况。观察到三种不同的表达模式：仅由刀豆蛋白A诱导、由刀豆蛋白A和白细胞介素2诱导以及由刀豆蛋白A和TCR刺激诱导。该方法对于分析T细胞分化途径和检测编码未识别细胞因子的cDNA克隆可能有用。
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