IMMUNE FUNCTION IN MICE LACKING THE PERFORIN GENE
IMMUNE FUNCTION IN MICE LACKING THE PERFORIN GENE
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DOI：
10.1073/pnas.91.23.10854
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发表时间：
1994-11-08
影响因子：
11.1
通讯作者：
CLARK, WR
中科院分区：
文献类型：
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作者：
WALSH, CM;MATLOUBIAN, M;CLARK, WR
缺乏穿孔素基因的小鼠是通过胚胎干细胞中的靶向基因破坏产生的。当感染淋巴细胞性脉络丛脑膜炎病毒 (LCMV) 时，无穿孔素 (-/-) 小鼠表现出明显的迹象，表明已基于 CD8 T 细胞的激活产生免疫反应，但无法清除 LCMV 感染。消除病毒的失败伴随着无法产生能够在体外裂解 LCMV 感染的成纤维细胞的脾细胞。感染 LCMV 的 -/- 小鼠的脾细胞在暴露于佛波醇 12-肉豆蔻酸酯 13-乙酸酯和离子霉素后能够裂解造血靶细胞，前提是靶细胞表达 Fas 抗原。来自-/-小鼠的脾细胞也通过胚细胞发生和增殖对混合白细胞培养物中的同种抗原作出反应。所得细胞能够裂解造血靶细胞，尽管不如以相同方式致敏的+/+同窝仔鼠的脾细胞。然而，仅当靶细胞表达Fas抗原时，才再次观察到-/-细胞的裂解。我们得出结论，无穿孔素-/-小鼠保留并表达体外表达的Fas裂解途径，但该途径不足以清除体内的LCMV感染。
Mice lacking the perforin gene were generated by using targeted gene disruption in embryonal stem cells. When infected with lymphocytic choriomeningitis virus (LCMV), perforin-less (-/-) mice showed clear signs of having mounted an immune response based on activation of CD8 T cells but were unable to dear the LCMV infection. This failure to eliminate virus was accompanied by a failure to generate spleen cells capable of lysing LCMV-infected fibroblasts in vitro. Spleen cells from LCMV-infected -/- mice were able to lyse hematopoietic target cells after exposure to phorbol 12-myristate 13-acetate and ionomycin, provided the target cells expressed the Fas antigen. Spleen cells from -/- mice also responded to alloantigen in mixed leukocyte culture by blastogenesis and proliferation. The resulting cells were able to lyse hematopoietic target cells, although not as well as spleen cells from +/+ littermates sensitized in the same manner. However, lysis by -/- cells was again seen only if the target cells expressed Fas antigen. We conclude that perforin-less -/- mice retain and express the Fas lytic pathway as expressed in vitro but that this pathway is insufficient to clear an LCMV infection in vivo.