Knight B C, Kissane S, Falciani F, Salmon M, Stanford M R, Wallace G R
Department of Ophthalmology, Rayne Institute, St Thomas' Hospital, London, UK.
J Neuroimmunol. 2006 Oct;179(1-2):126-31. doi: 10.1016/j.jneuroim.2006.06.002. Epub 2006 Aug 24.
To investigate changes in immune response genes following Toxoplasma gondii infection of Müller cells.
Human Müller cells were infected or mock infected with two strains of T. gondii (RH and Prugniaud). RNA and supernatants were collected from infected and uninfected cells at 2 and 24 h. RNA from the two time points were compared using a custom made DNA microarray. Real time PCR or human cytokine antibody array was used to confirm up-regulation of immune molecules.
Gene expression in infected cells showed up-regulation of CCL2, IL-6, CXCL8, and CXCL2. CCL2 and CXCL2 gene expression was confirmed by real time PCR. IL-6 and CXCL8 protein production was confirmed by a cytokine antibody array. IL-4 production was observed by cytokine antibody array but not by DNA microarray. In contrast, infection with T. gondii did not induce interferon-gamma (IFNgamma) and IL-12 expression, molecules conventionally associated with the inter-conversion of tachyzoite to bradyzoite.
These results indicate that while in vitro infected Müller cells may be capable of inducing an immune response by attracting blood-borne leucocytes, they do not appear able to directly control the proliferation of T. gondii.
研究弓形虫感染穆勒细胞后免疫反应基因的变化。
用人的穆勒细胞分别感染或模拟感染两株弓形虫(RH株和Prugniaud株)。在感染后2小时和24小时从感染和未感染的细胞中收集RNA和上清液。使用定制的DNA微阵列比较两个时间点的RNA。采用实时定量聚合酶链反应或人细胞因子抗体阵列来确认免疫分子的上调。
感染细胞中的基因表达显示CCL2、IL-6、CXCL8和CXCL2上调。通过实时定量聚合酶链反应确认了CCL2和CXCL2基因表达。通过细胞因子抗体阵列确认了IL-6和CXCL8蛋白的产生。通过细胞因子抗体阵列观察到了IL-4的产生,但DNA微阵列未检测到。相反,弓形虫感染未诱导干扰素-γ(IFNγ)和IL-12的表达,而这两种分子通常与速殖子向缓殖子的相互转化有关。
这些结果表明,虽然体外感染的穆勒细胞可能能够通过吸引血源性白细胞诱导免疫反应,但它们似乎无法直接控制弓形虫的增殖。