Imunização de camundongos BALB/C com as proteínas recombinantes TSA, LelF e STI de Leishmania (Viannia) braziliensis e avaliação da eficácia da vacina
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2016-03-04
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Universidade Federal de Goiás
Resumo
In Brazil, L. (V.) braziliensis is specie responsible for many cases of American
Tegumentary Leishmaniasis (ETL). To effective vaccine development, formulations
contained immunogenic antigens associated with adjuvants that induce a T helpe 1
(Th1) immune response are been investigated, among of antigen have the “Thiol
Specific Antioxidant” (rTSA), “Stress Inducible protein 1” (rSTI) and “Leishmania
elongation initiation fator” (rLeIF) of L. major, which have been immunogenic and
promising. The genes that corresponding to these antigens in L. (V.) braziliensis were
sequenced and recombinants proteins were obtained and used in this study. Works that
used BCG and β-glucan showed that these could provide protection against not related
pathogens. The present study had as aim to evaluate the immunogenicity and the
effectiveness of the vaccine using the rTSA, rSTI and rLeIF proteins (Triple),
associated or not with β-glucan, with or without BCG stimulus. BALB/c mice were
divided in 2 groups, one of the groups was stimulated with BCG and the other not.
Posteriorly, the animals were divided in subgroups and vaccinated with triple,
associated or not with β-glucan. Mice’s serum were collected to search of cytokines and
antibodies by immunoenzymatic technique. Lymph node and spleen’s cells were
cultivated and cytokines searched in supernatants. BCG stimulation induced a greater
IFN-γ production. The vaccinated mice with triple produced IgG total, IgG1 and IgG2a
specifics to L. (V.) braziliensis. BCG stimulation induced a high IgG total and IgG2a
production in animals that received the triple without β-glucan. To evaluate the
protection provided by vaccine, the immunized animals were infected with L. (V.)
braziliensis and paw them measured along of 8 weeks. The subgroup don’t stimulated
with BCG and vaccinated with triple, associated with β-glucan, has developed bigger
lesions that the others subgroups. The subgroups that received only BCG stimulation or
BCG stimulation and vaccine with triple plus β-glucan presented bigger lesions that
animal don’t stimulated or vaccinated. After infection by L. (V.) braziliensis, (i) the
animals stimulated with BCG and vaccinated with triple presents more IgG specific to
L. (V.) braziliensis; (ii) the immunizations with triple maintained IFN-γ levels elevated;
(iii) all animals presented lower IL-17 levels in serum; (iv) the subgroups produced
similar IFN-γ levels, but with a high IL-17 production in subgroup vaccinated with
triple plus β-glucan, don’t BCG stimulated, in spleen; (v) the animals that received only
BCG produced more IFN-γ and IL-17 in lymph node. In conclusion: the immunizations
with triple were immunogenic; the immunization scheme containing triple plus β-
glucan, without BCG stimulation, provided partial protection against L. (V.) braziliensis
infection; BCG stimulations induced biggest lesions in subgroup that received only
saline or triple plus β-glucan; the vaccination scheme containing triple maintained high
IFN-γ production after L. (V.) braziliensis infection; in infection site, the BCG
stimulation induced a Th1/Th17 heterologous response.
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MATOS, G. G. Imunização de camundongos BALB/C com as proteínas recombinantes TSA, LelF e STI de Leishmania (Viannia) braziliensis e avaliação da eficácia da vacina. 2016. 133 f. Dissertação (Mestrado em Medicina Tropical e Saúde Publica) - Universidade Federal de Goiás, Goiânia, 2016.