TNP-specific Lyt-2+ cytolytic T cell clones preferentially respond to TNP-conjugated epidermal cells.
TNP-specific Lyt-2+ cytolytic T cell clones preferentially respond to TNP-conjugated epidermal cells.
复制标题
TNP 特异性 Lyt-2 溶细胞 T 细胞克隆优先响应 TNP 结合的表皮细胞。
作者：
S. Shimada;S. Katz
诱导半抗原特异性变应性接触敏感性（CS）的最有效方法是通过表皮应用半抗原。另一种有效的方法是通过皮下施用半抗原化的表皮细胞（EC）。后一种方法比皮下施用半抗原化脾细胞（SC）诱导更强烈和更持久的CS。因此，可能有一些独特的EC，当半抗原化，使他们能够产生效应细胞更有效地比SC。因此，我们试图产生T细胞克隆，半抗原和表皮特异性。用7%三硝基氯苯涂抹小鼠后4天，获得引流淋巴结细胞并纯化T细胞。将这些细胞与三硝基苯基化（TNP）朗格汉斯细胞富集EC共培养。4天后，收获细胞并置于非TNP缀合的EC上。将细胞再刺激并静置三次，然后通过加入白细胞介素2的有限稀释进行克隆，然后继续加入白细胞介素2。通过[3 H]-胸苷掺入来评估T细胞的增殖。细胞毒性测定利用TNP缀合的伴刀豆球蛋白A SC胚细胞或EC作为靶标。克隆A-2和E-4是Thy-1+、Lyt-2+和L3 T4-以及TNP特异性的。与非克隆的TNP特异性T细胞相反，克隆优先增殖响应TNP-EC，而不是TNP-SC。也与非克隆的T细胞相反，克隆优先对TNP-EC的细胞毒性;与TNP-SC相比，当TNP-EC被用作靶时，杀伤增加了8至32倍。因此，克隆A-2和E-4表现出半抗原和表皮特异性。被识别的表皮特异性表位是未知的，但遗传限制和抗体抑制研究表明，它与H-2K共识别。
A most effective method for the induction of hapten-specific allergic contact sensitivity (CS) is via epicutaneous application of the hapten. Another effective method is by the administration of haptenated epidermal cells (EC) subcutaneously. The latter method induces more intense and longer lasting CS than does the subcutaneous administration of haptenated spleen cells (SC). Thus, there may be something unique about EC which, when haptenated, allows them to generate effector cells more effectively than do SC. We therefore attempted to generate T cell clones that were both hapten- and epidermal-specific. Four days after painting mice with 7% trinitrochlorobenzene, draining lymph node cells were obtained and T cells were purified. These cells were co-cultured with trinitrophenylated (TNP) Langerhans cell-enriched EC. After 4 days, cells were harvested and rested on non-TNP-conjugated EC. The cells were restimulated and rested three times, and were then cloned by limiting dilution with added interleukin 2, which was then continually added. Proliferation of T cells was assessed by [3H]-thymidine incorporation. Cytotoxicity assays utilized TNP-conjugated concanavalin A SC blasts or EC as targets. Clones A-2 and E-4 are Thy-1+, Lyt-2+, and L3T4-, and TNP-specific. In contrast to noncloned TNP-specific T cells, the clones proliferate preferentially in response to TNP-EC rather than TNP-SC. Also in contrast to noncloned T cells, the clones were preferentially cytotoxic for TNP-EC; compared to TNP-SC, there was an eight- to 32-fold increase in killing when TNP-EC were used as targets. Clones A-2 and E-4 therefore exhibit hapten and epidermal specificity. The epidermal-specific epitope that is recognized is unknown, but genetic restriction and antibody inhibition studies indicate that it is co-recognized with H-2K.