PARTICIPATION OF TARGET FAS PROTEIN IN APOPTOSIS PATHWAY INDUCED BY CD4(+) TH1 AND CD8(+) CYTOTOXIC T-CELLS
PARTICIPATION OF TARGET FAS PROTEIN IN APOPTOSIS PATHWAY INDUCED BY CD4(+) TH1 AND CD8(+) CYTOTOXIC T-CELLS
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DOI：
10.1073/pnas.91.10.4185
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发表时间：
1994-05-10
影响因子：
11.1
通讯作者：
MARSHAKROTHSTEIN, A
中科院分区：
文献类型：
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作者：
JU, ST;CUI, HL;MARSHAKROTHSTEIN, A
本研究结果表明，靶细胞中Pas蛋白的存在对于克隆的CD 4（+）Th 1细胞介导的细胞毒性（导致细胞凋亡）是必不可少的。使用丝裂原活化的B细胞作为靶细胞，观察到MRL/MpJ+靶细胞对CD 4（+）Th 1效应细胞的抗原依赖性裂解，但MRL/MpJ-lpr靶细胞则没有。MRL/MpJ-lpr同源株Pas表达缺陷。来自C3 H.MRL-lpr小鼠的各种淋巴组织的靶细胞也对Th 1效应物的凝集素依赖性细胞毒性具有抗性，而C3 H/HeJ靶是敏感的。此外，Cr-51释放前的快速DNA片段化仅在C3 H/HeJ靶中诱导。因此，由Th 1效应子诱导的细胞毒性与靶Fas表达相关。与Th 1效应物相反，CD 8（+）细胞毒性T淋巴细胞（CTL）杀死C3 H.MRL-lpr靶。当在EGTA和MgCl 2（其螯合细胞外Ca 2 + [（Ca 2+）（ext）]）存在下测定细胞毒性时，仅C3 H.MRL-lpr靶标变得对CD 8（+）CTL具有抗性。Th 1和CD 8（+）效应细胞的这种（Ca 2+）（ext）非依赖性细胞毒性可以用未标记的C3 H/HeJ胸腺细胞或携带鼠Fas-人mu基因构建体的转染瘤抑制。相比之下，C3 H.MRL-lpr胸腺细胞和未转染的亲本细胞系是较差的抑制剂。我们的研究表明，CD 4（+）Th 1细胞和CD 8（+）CTL在它们的（Ca 2+）（ext）依赖性细胞毒性方面不同，但共享（Ca 2+）（ext）非依赖性细胞毒性，其需要Fas分子参与细胞毒性信号转导，导致靶细胞凋亡。
The results presented here provide evidence that the presence of Pas protein in target cells is essential to permit cytotoxicity (resulting in apoptosis) mediated by cloned CD4(+) Th1 cells. Using mitogen-activated B cells as targets, antigen-dependent lysis by CD4(+) Th1 effecters was observed with MRL/MpJ+ but not with MRL/MpJ-lpr targets. The congenic MRL/MpJ-lpr strain is defective in Pas expression. Target cells from various lymphoid tissues of C3H.MRL-lpr mice were also resistant to the lectin-dependent cytotoxicity of Th1 effecters, whereas C3H/HeJ targets were sensitive. Moreover, a rapid DNA fragmentation prior to Cr-51 release was induced only in C3H/HeJ targets. Thus, cytotoxicity induced by Th1 effecters correlates with target Fas expression. In contrast to Th1 effectors, CD8(+) cytotoxic T lymphocytes (CTLs) killed C3H.MRL-lpr targets. When cytotoxicity was assayed in the presence of EGTA and MgCl2, which chelates extracellular Ca2+ [(Ca2+)(ext)], only C3H.MRL-lpr targets became resistant to CD8(+) CTLs. This (Ca2+)(ext)-independent cytotoxicity of both Th1 and CD8(+) effecters could be inhibited with unlabeled C3H/HeJ thymocytes or with a transfectoma carrying a murine Fas-human mu gene construct. In comparison, C3H.MRL-lpr thymocytes and the nontransfected parental cell line were poor inhibitors. Our study demonstrates that CD4(+) Th1 cells and CD8(+) CTLs differ in their (Ca2+)(ext)-dependent cytotoxicity but share a (Ca2+)(ext)-independent cytotoxicity that requires participation of Fas molecules for cytotoxic signal transduction leading to target apoptosis.