Verdon R, Keusch G T, Tzipori S, Grubman S A, Jefferson D M, Ward H D
Department of Medicine, New England Medical Center, Boston, Massachusetts 02111, USA.
J Infect Dis. 1997 May;175(5):1268-72. doi: 10.1086/593695.
Cryptosporidium parvum infection in the immunosuppressed host is frequently complicated by biliary tract involvement. The recent production of human biliary epithelial cell lines was exploited to develop an in vitro model of biliary cryptosporidiosis. Infection with C. parvum oocysts was detected by IFA and ELISA and confirmed by transmission electron microscopy. Inoculation of monolayers with 10(4) to 5 X 10(5) oocysts/well resulted in a dose-dependent increase in infection. Time-course experiments showed that the number of parasitic stages was maximal at 18-24 h after inoculation. Infection was significantly enhanced by bile at concentrations of 50 and 100 microg/mL and inhibited by 400 microg/mL paromomycin. Infection of human biliary cells with C. parvum can be consistently achieved and monitored by use of IFA or ELISA. This system will be of use in evaluating mechanisms of C. parvum infection and response to therapeutic agents in biliary cryptosporidiosis.
免疫抑制宿主中的微小隐孢子虫感染常因胆道受累而复杂化。利用最近建立的人胆管上皮细胞系开发了一种胆道隐孢子虫病的体外模型。通过免疫荧光分析（IFA）和酶联免疫吸附测定（ELISA）检测微小隐孢子虫卵囊感染情况，并通过透射电子显微镜进行确认。每孔接种10⁴至5×10⁵个卵囊的单层细胞，感染呈剂量依赖性增加。时间进程实验表明，接种后18 - 24小时寄生虫阶段数量最多。50和100微克/毫升浓度的胆汁显著增强感染，而400微克/毫升的巴龙霉素则抑制感染。使用IFA或ELISA可以持续实现并监测微小隐孢子虫对人胆管细胞的感染。该系统将用于评估微小隐孢子虫感染机制以及胆道隐孢子虫病对治疗药物的反应。
