THE HUMAN NATURAL-KILLER-CELL RECEPTOR FOR MAJOR HISTOCOMPATIBILITY COMPLEX CLASS-I MOLECULES - SURFACE MODULATION OF P58 MOLECULES AND THEIR LINKAGE TO CD3 ZETA-CHAIN, FC-EPSILON-RI GAMMA-CHAIN AND THE P56(LCK) KINASE
THE HUMAN NATURAL-KILLER-CELL RECEPTOR FOR MAJOR HISTOCOMPATIBILITY COMPLEX CLASS-I MOLECULES - SURFACE MODULATION OF P58 MOLECULES AND THEIR LINKAGE TO CD3 ZETA-CHAIN, FC-EPSILON-RI GAMMA-CHAIN AND THE P56(LCK) KINASE
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DOI：
10.1002/eji.1830241040
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发表时间：
1994-10-01
影响因子：
5.4
通讯作者：
MORETTA, A
中科院分区：
文献类型：
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作者：
BOTTINO, C;VITALE, M;MORETTA, A
由GL 183和EB 6单克隆抗体（mAb）识别的自然杀伤细胞（NK）特异性p58表面分子已被证明代表HLA-C分子的推定NK受体。p58受体和HLA-C之间的相互作用导致NK介导的靶细胞裂解的抑制。在这项研究中，GL 183（-）EB 6（+）克隆（Cw 4特异性），在单克隆抗体诱导的EB 6分子的表面调节后，获得了裂解Cw 4（+）C1 R细胞的能力。在共表达GL 183和EB 6分子并且不能杀死Cw 3保护的靶细胞的NK克隆中，mAb诱导的EB 6分子的调节导致GL 183分子的选择性共调节和Cw 3转染的P815鼠细胞的裂解。与共调节实验一致，我们还表明，在毛地黄皂苷存在下，细胞裂解后，GL 183和EB 6分子可以从GL 183（+）/EB 6（+）克隆中共免疫沉淀。p58受体也揭示了与属于zeta家族的分子的关联（i. e. CD 3 ζ和FceRI γ链）。从多克隆激活的p58（+）NK细胞免疫沉淀的p58复合物的二维对角凝胶分析表明，以共价连接的zeta-zeta同源二聚体或zeta-gamma异源二聚体的形式优先与CD 3 zeta链结合，而gamma-gamma同源二聚体的含量较低。然而，也可以分离出与γ-γ同源二聚体具有独特关联的p58（+）克隆。用抗p56（lck）抗体探测免疫沉淀的p58复合物也揭示了与src家族成员的关联。此外，通过p58分子的NK克隆的mAb介导的信号传导诱导p58/p56（lck）缔合的增加。然而，在诱导CD 16相关CD 3 ζ链的最佳体内酪氨酸磷酸化的相同实验条件下，在p58相关CD 3 ζ链中未检测到酪氨酸磷酸化。在这些体内实验中，抗CD 16和抗p58 mAb均不能诱导γ链的酪氨酸磷酸化。最后，抗p58介导的通过CD 16分子触发的NK细胞抑制并不伴随着CD 16相关的CD 3 zeta链的酪氨酸磷酸化的下调。
The natural killer cell (NK)-specific p58 surface molecules, recognized by the GL183 and EB6 monoclonal antibodies (mAb), have been shown to represent the putative NK receptor for HLA-C molecules. The interaction between p58 receptors and HLA-C results in inhibition of the NK-mediated target cell lysis. In this study, GL183(-)EB6(+) clones (Cw4-specific), after mAb-induced surface modulation of EB6 molecules, acquired the ability to lyse the Cw4(+) C1R cells. In NK clones co-expressing both GL183 and EB6 molecules and unable to kill Cw3-protected target cells, the mAb-induced modulation of EB6 molecules resulted both in selective co-modulation of GL183 molecules and in the lysis of Cw3-transfected P815 murine cells. In Line with the co-modulation experiments we also show that the GL183 and EB6 molecules can be co-immunoprecipitated from GL183(+)/EB6(+) clones after cell lysis in the presence of digitonin. The p58 receptor also revealed an association with molecules belonging to the zeta family (i. e. CD3 zeta and FceRI gamma chains). Two-dimensional diagonal gel analysis of the p58 complex immunoprecipitated from polyclonally activated p58(+) NK cells indicated a preferential association with CD3 zeta chains either in the form of covalently linked zeta-zeta homodimers or in the form of zeta-gamma heterodimers, while gamma-gamma homodimers were detectable in low amounts. However, p58(+) clones displaying a unique association with gamma-gamma homodimers could also be isolated. Probing the immunoprecipitated p58 complex with anti-p56(lck) antibody also revealed an association with this member of the src family. In addition, mAb-mediated signaling of NK clones via p58 molecules induced increments of p58/p56(lck) association. However, under the same experimental conditions that induced optimal in vivo tyrosine phosphorylation of the CD16-associated CD3 zeta chains, no tyrosine phosphorylation was detected in the p58-associated CD3 zeta; chains. In these in vivo experiments neither anti-CD16 nor anti-p58 mAb could induce tyrosine phosphorylation of the gamma chains. Finally, the anti-p58-mediated inhibition of the NK cell triggering via CD16 molecules was not accompained by a down-regulation of the tyrosine phosphorylation of the CD16-associated CD3 zeta chains.