Cottrell B, Pye C, Butterworth A
Department of Pathology, University of Cambridge, UK.
Parasite Immunol. 1989 Jan;11(1):91-104. doi: 10.1111/j.1365-3024.1989.tb00651.x.
Human peripheral blood monocytes from normal donors were isolated by differential centrifugation and cultured in vitro in hydrophobic Teflon-coated tissue culture bags. Cells were harvested between 0 and 10 days and tested for their ability to kill schistosomula of Schistosoma mansoni in an in-vitro cytotoxicity assay. Freshly isolated, unstimulated monocytes demonstrated minimal cytotoxic capability. However, this was increased if the cells were pretreated with human recombinant gamma interferon (IFN-gamma), or with specific anti-S. mansoni antiserum. As the monocytes matured in vitro there were marked increases in the levels of antibody-independent killing of schistosomula. Monocytes grown in vitro with IFN-gamma (10(4) u/ml) took 2-3 days to develop almost maximal cytotoxicity (mean 94% kill of schistosomula). In contrast, unstimulated monocytes (no IFN-gamma) took between 5 and 7 days to achieve comparable cytotoxicity (mean 99% kill). Killing of the schistosomula was dependent upon a high effector to target ratio, and was a relatively slow phenomenon in vitro, parasite attrition occurring between 17 and 36 h. Supernatants from cytotoxic macrophages were ineffective in mediating cytotoxicity of the parasite.
通过差速离心法从正常供体中分离出人外周血单核细胞,并在疏水性聚四氟乙烯涂层的组织培养袋中进行体外培养。在0至10天之间收获细胞,并在体外细胞毒性试验中测试其杀伤曼氏血吸虫童虫的能力。新鲜分离的未受刺激的单核细胞表现出最小的细胞毒性能力。然而,如果细胞用人重组γ干扰素(IFN-γ)或特异性抗曼氏血吸虫抗血清进行预处理,细胞毒性会增加。随着单核细胞在体外成熟,对童虫的非抗体依赖性杀伤水平显著增加。在体外与IFN-γ(10⁴u/ml)一起培养的单核细胞需要2至3天来发展出几乎最大的细胞毒性(平均94%的童虫杀伤率)。相比之下,未受刺激的单核细胞(无IFN-γ)需要5至7天才能达到相当的细胞毒性(平均99%的杀伤率)。对童虫的杀伤取决于高效应细胞与靶细胞比例,并且在体外是一个相对缓慢的现象,寄生虫损耗发生在17至36小时之间。细胞毒性巨噬细胞的上清液在介导寄生虫的细胞毒性方面无效。