Zaalouk, T.K.; McDonald, V.; Blanshard, C.
April 2003
Gut;Apr2003 Supplement 1, Vol. 52, pA100
Academic Journal
Introduction: E intestinalis, a microsporidian pathogen in man, has emerged as a serious opportunistic ,pathogen in HIV infected individuals. in this study the development of the parasite in different cell lines was investigated. In addition, the effect of pro-inflammatory cytokines on this development was analysed. Methods: Seventy percent confluent monolayers of human (HT-29, Caco-2) colonic and murine (CMT-93) rectal adenocarcinoma cell lines were incubated with 4×10[sup 5] E intestinalis spores at 37°C and the cells were examined for intracellulor parasitic development after 24, 48, and 72h. In cytokine experiment cells were cultured in the presence of the cytokine(s) for 24 h prior to inoculation of E. intestinalis, and for a further 72h after inoculation in a concentration that was previously found not to damage the cells. Results: Our results showed E intestinalis was able to establish considerable infection in CMT-93 but, surprisingly, not in the human cell lines. The different stages of intracellular parasitic development were clearly visible in the CMT-93 cell line, making it a suitable in vitro model to study the infection in intestinal epithelium. Our results showed even small concentrations of IFN-α inhibited the development of the parasite by more than 90%. TNF-α and IL-6 also significantly reduced the infection and a combination of both cytokines had significantly higher inhibitory effect than either cytokine alone, suggesting that they have a synergistic effect. Conclusions: Observations from this study suggest that the CMT-93 cell line is appropriate for use in an in vitro model of E intestinalis infection. The requirement for IFN-α indicates that a Th1 response is important in control of infection. Additionally, enterocyte derived cytokines TNF-α and IL-6 may have an important role in intestinal immunity to infection.


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