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