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Contributed Papers: Posters
Immunology


Cryptosporidium parvum cellular recruitment during the inflammatory response in mice


Bozzoli, L.M1, Sakai,Y.I2, Borges, M 3, Neto, A.P2, De Gaspari, E.N1.
1- Seção de Imunologia, Instituto Adolfo Lutz 2-Seção de Patologia, Instituto Adolfo Lutz 3-Laboratório de Imunoquímica, Instituto Butantan São Paulo/SP. egaspari@ial.sp.gov.br

C. parvum is a common opportunistic infection associated with HIV/AIDS. Infection by this organism triggers a complex array of innate and cell-mediated immune responses within the intestinal mucosa. T-helper type 1 cytokines, and particularly IFN-g, have long been considered to be the main orchestrators of the immune responses to this infection, but recent studies suggest that T helper type 2 cytokines may also be involved. We have it was previously demonstrated that B10A mice (H-2a) were more resistant to C.parvum infection and produced high level of oocysts shedding (20-fold higher) as compared to C57BL/6(H-2b), BALB/c(H-2d), and A/Sn (H-2a) strains after (i.g.) infection. To better understand the immunoregulatory phenomena of infection of C.parvum, the type 2, IL-4, IL-5 and IL-10 cytokine profiles of mice were studied . Mice were infected with 1 dose of 5x106 parasites (waterborne). On the 7th day, exudates were centrifuged, supernatants were removed for cytokines analysis, and cells pellets were re-suspended in 1.0 mL PBS and counted in Fuchs-Rosenthal double-ruling chamber. Two hundred thousand cells were centrifuged onto microscope slides at 500 rpm for 5 min using a cytospin centrifuge. The slides were stained by modified papanicolaou method, and the presence of the C.parvum was determined by modified Romanowsky stain. The percentages of macrophages, leucocytes, eosinophils, monocytes and lymphocytes were also determined. IFN-g, L-4, IL-5, and IL-10 levels were measured by capture ELISA. The parasites induced IFN-g in all analyzed strains. In contrast, no IL-4 response was detected. Taken together, these results demonstrated higher production of Th 1 cytokines in all analyzed strains, whereas IFN-g was higher in B10A.The best pattern of Th 2 cytokines was observed in the strains producing IL-5 (B10A), IL-10 (C57Bl/6), as compared to lower production of IFN-g (BALB/c), IL-5 (C57BL/6) and IL10 (B10A). A monoclonal antibody against C.parvum using i.g. immunization was produced.

Financial support: FAPESP



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