Silva J S, Vespa G N, Cardoso M A, Aliberti J C, Cunha F Q
Department of Immunology, Faculty of Medicine of Ribeirão Preto, São Paulo, Brazil.
Infect Immun. 1995 Dec;63(12):4862-7. doi: 10.1128/iai.63.12.4862-4867.1995.
Cell invasion by Trypanosoma cruzi and its intracellular replication are essential for continuation of the parasite life cycle and for production of Chagas' disease. T. cruzi is able to replicate in nucleated cells and can be killed by activated macrophages. Gamma interferon (IFN-gamma) is one of the major stimuli for the activation of macrophages and has been shown to be a key activation factor for the killing of intracellular parasites through a mechanism dependent upon nitric oxide (NO) biosynthesis. We show that although the addition of exogenous tumor necrosis factor alpha (TNF-alpha) does not potentiate the trypanocidal activity of IFN-gamma in vitro, treatment of resistant C57BI/6 mice with an anti-TNF-alpha monoclonal antibody increased parasitemia and mortality. In addition, the anti-TNF-alpha-treated animals had decreased NO production, both in vivo and in vitro, suggesting an important role for TNF-alpha in controlling infection. In order to better understand the role of TNF-alpha in the macrophage-mediating killing of parasites, cultures of T. cruzi-infected macrophages were treated with an anti-TNF-alpha monoclonal antibody. IFN-gamma-activated macrophages failed to kill intracellular parasites following treatment with 100 micrograms of anti-TNF-alpha. In these cultures, the number of parasites released at various time points after infection was significantly increased while NO production was significantly reduced. We conclude that IFN-gamma-activated macrophages produce TNF-alpha after infection by T. cruzi and suggest that this cytokine plays a role in amplifying NO production and parasite killing.
克氏锥虫的细胞侵袭及其细胞内复制对于寄生虫生命周期的延续以及恰加斯病的产生至关重要。克氏锥虫能够在有核细胞中复制,并且可被活化的巨噬细胞杀死。γ干扰素(IFN-γ)是激活巨噬细胞的主要刺激因子之一,并且已被证明是通过依赖于一氧化氮(NO)生物合成的机制杀死细胞内寄生虫的关键激活因子。我们发现,尽管在体外添加外源性肿瘤坏死因子α(TNF-α)不会增强IFN-γ的杀锥虫活性,但用抗TNF-α单克隆抗体治疗抗性C57BI/6小鼠会增加寄生虫血症和死亡率。此外,抗TNF-α处理的动物在体内和体外的NO产生均减少,这表明TNF-α在控制感染中起重要作用。为了更好地理解TNF-α在巨噬细胞介导的寄生虫杀伤中的作用,用抗TNF-α单克隆抗体处理克氏锥虫感染的巨噬细胞培养物。在用100微克抗TNF-α处理后,IFN-γ激活的巨噬细胞无法杀死细胞内寄生虫。在这些培养物中,感染后各个时间点释放的寄生虫数量显著增加,而NO产生显著减少。我们得出结论,IFN-γ激活的巨噬细胞在被克氏锥虫感染后产生TNF-α,并表明这种细胞因子在放大NO产生和寄生虫杀伤中起作用。