Dyugovskaya L, Ginsburg H
Department of Immunology, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.
Cell Immunol. 1996 Apr 10;169(1):30-9. doi: 10.1006/cimm.1996.0087.
Rat and mouse fibroblast monolayers were infected with murine cytomegalovirus (MCMV) by 2 hr incubation to allow virus to penetrate the cells. Then lymph node cells (LNC) from rats infected with MCMV and from untreated rats were added together with human recombinant interleukin-2 (hrIL-2). Cytopathic plaques appeared within 2-3 days. In culture of fibroblasts only, 30-40 plaques per well progressed into confluent cytopathy within 6-8 days. In cultures with LNC syngeneic to fibroblasts, plaques appeared; however, the cytotoxic T lymphocyte population that developed and specific apoptotic fragmentation eliminated the cytomegalic cells in the plaques. The surrounding cells stretched to the area, the cytopathic plaques disappeared, and the monolayer resumed its uninfected texture. No plaque-forming units could be isolated from such cured cultures. In allogeneic combination there was no apoptotic target cell killing. However, in cultures stimulated by hrIL-2, plaque growth was arrested and the plaques remained rudimentary. Similarly, arrest of plaques was also obtained in cultures containing LNC from uninfected rats, but only if stimulated by hrIL-2. In mouse fibroblasts carrying the rat LNC, plaque growth was not arrested, and the culture developed into confluent cytopathy. Interferon (IFN)-gamma or -alpha,beta added 24 hr before and 2 hr after infection abolished plaque appearance or arrested growth. IFN-gamma appeared to be the most effective. Fluids harvested from cured cultures also protected from plaque development. Antibodies to IFN-gamma, but not to IFN-alpha,beta, neutralized this capacity in the culture fluids. It is concluded that IFN-gamma produced by the LNC played a major role in the cure of the fibroblast culture from MCMV infection. A mechanism of cell-mediated immunity operating in resolving virus infection is proposed.
将大鼠和小鼠成纤维细胞单层培养物与鼠巨细胞病毒（MCMV）共同孵育2小时，以使病毒能够穿透细胞。然后，将感染MCMV的大鼠和未处理大鼠的淋巴结细胞（LNC）与人重组白细胞介素-2（hrIL-2）一起加入。在2-3天内出现细胞病变斑块。仅在成纤维细胞培养物中，每孔30-40个斑块在6-8天内发展为融合性细胞病变。在与成纤维细胞同基因的LNC培养物中，出现了斑块；然而，发育形成的细胞毒性T淋巴细胞群体和特异性凋亡碎片消除了斑块中的巨细胞。周围细胞伸展到该区域，细胞病变斑块消失，单层恢复其未感染时的质地。从这种治愈的培养物中无法分离出斑块形成单位。在异基因组合中，没有凋亡靶细胞的杀伤作用。然而，在hrIL-2刺激的培养物中，斑块生长受到抑制，斑块仍处于初级阶段。同样，在含有未感染大鼠LNC的培养物中也观察到斑块生长受到抑制，但前提是受到hrIL-2刺激。在携带大鼠LNC的小鼠成纤维细胞中，斑块生长未受到抑制，培养物发展为融合性细胞病变。在感染前24小时和感染后2小时添加干扰素（IFN）-γ或-α，β可消除斑块出现或抑制其生长。IFN-γ似乎最有效。从治愈的培养物中收获的液体也能防止斑块形成。针对IFN-γ而非IFN-α，β的抗体可中和培养液中的这种能力。得出的结论是，LNC产生的IFN-γ在成纤维细胞培养物从MCMV感染中治愈的过程中起主要作用。提出了一种在解决病毒感染中起作用的细胞介导免疫机制。
