cDNA microarray analysis of toxoplasmic retinochoroiditis

利用正常小鼠mRNA构建的15000个克隆组成的cDNA微阵列系统，对刚地弓形虫感染诱导基因进行了分析。5个基因组，β -微球蛋白，DMR-N9，斑点14，睾丸素，和一个未知的（暂时命名为T.g.Res）。X)被接走了。通过比较WT耐药菌株（BALB/c）和WT敏感菌株（C57BL/6）以及WT和IFN-γ敲除（GKO） BALB/c小鼠的mRNA表达情况，发现T.g.Res.X在弓形虫感染的耐药菌株中表达。WT C57BL/6、WT BALB/c和GKO小鼠经口感染弓形虫囊。分析方法包括眼科检查、荧光素血管造影（FA）、定量竞争聚合酶链反应（QC-PCR）、IFN-γ和分期转化标记物的逆转录(RT)-PCR和cDNA微军检测。在WT C57BL/6小鼠中，弓形虫的分布顺序为：大脑、视网膜、脉络膜、巩膜末端视神经（ON）。后视网膜中弓形虫的载量最高，远高于WT BALB/c小鼠。GKO小鼠弥散性感染明显，脉络膜和ON中弓形虫载量最高。感染后，WT C57BL/6小鼠的IFN-γ mRNA表达高于WT BALB/c小鼠。GKO小鼠存在速殖子，WT C57BL/6小鼠存在慢殖子。FAG显示GKO小鼠视网膜毛细血管有染料渗漏。与耐药WT菌株不同，敏感WT菌株视网膜内弓形虫载量持续增加。IFN-γ可调节眼内弓形虫负荷和相互转化。用GKO小鼠建立弓形虫性血管炎模型，采用定量定量pcr和FA检测系统。利用基因芯片系统确定弓形浆性视网膜脉络膜炎相关基因的实验正在进行中

By using our own made cDNA microarray system composed with around 15,000 clones established by using mRNA from normal mice, Toxoplasma gondii-infection-induced genes were analyzed.Five gene groups, beta2-microglobulin, DMR-N9, spot 14,teststeron, and an unknown (tentatively named as T.g.Res.X) were picked up. T.g.Res.X was shown to express in a resistant strain to T.gondii infection when the mRNA expression was examined by comparing between a WT resistant strain (BALB/c) and a WT susceptible strain (C57BL/6) and also comparing between WT and IFN-γ knockout (GKO) BALB/c mice.WT C57BL/6, WT BALB/c, and GKO mice were infected with cysts of T.gondii perorally. Analysis included ophthalmic examinations, fluorescein angiography (FA), a quantitative competitive polymerase chain reaction (QC-PCR) assay, reverse transcription (RT)-PCR for IFN-γ and stage conversion markers and cDNA microarmy assayIn WT C57BL/6 mice, T.gondii was detected in the order Of the brain, retina, choroid, sclera end optic nerve (ON). The highest T.gondii load was observed in the posterior retina, and was much greater than that in WT BALB/c mice. In GKO mice, disseminated infection was evident and the T.gondii load was highest in the choroid, and ON. IFN-γ mRNA expression in WT C57BL/6 mice was higher than that in WT BALB/c mice after infection. Tachyzoites existed in GKO mice, while bradyzoites existed in WT C57BL/6 mice FAG showed dye leakage from the retinal capillaries of GKO mice.The T.gondii load in the retina in the susceptible WT strain continued to increase, unlike in the resistant WT strain. IFN-γ was shown to regulate the T gondii load and interconversion in the eye. A toxoplasmic vasculitis model was established with GKO mice and assay systems with QC-PCR and FA.Experiments are now under way to define the toxoplasmic retinochoroiditis related genes by using cDNA microarray system