A cell fractionation procedure previously developed for Trypanosoma cruzi was applied to isolated the plasma membrane of promastigotes of Leishania mexicana amazonensis. The cell, swollen in an hypotonic mediun, were disrupted in the presence of a nonionic detergent and the membrane fraction isolated by differencial centrifugation. Electron microscopy showed that the fraction consisted of pieces of the plasma membrane associated with subpellicular microtubules. It was also shown that this fraction is able to induce cell-mediated immune response in mice.
The immunogenic and protective activity of an extract of S. mansoni, obtained by incubation of viable adult worms in buffered saline, was evaluated in rabbits and mice. Animal immunization with this extract resulted in the development of both humoral and cellular immune response. All immunized rabbits developed high levels (91 to 100%) of cytotoxic antibodies as determined by in vitro assays of cytotoxic activity of their sera against viable schistosomules. Immunized animals challenged with S. mansoni cercariae showed a lower parasite load than that of normal controls. Protective activity was 88.6% and 54.0% in immunized rabbits and mice, respectively.
Avaliou-se em coelhos e camundongos, a atividade imunogênica e protetora de um extrato antigênico de Schistosoma mansoni, obtido pela estocagem de vermes adultos em solução salina tamponada (Extrato Salino). A imunização dos animais determinou o desenvolvimento de resposta imune celular e humoral, avaliada por provas específicas. Todos os coelhos imunizados com ES, desenvolveram altos níveis de anticorpo citotóxico (91 a 100%), determinados pela avaliação da atividade citotóxica in vitro, contra esquitossômulos. Concluiu-se que os coelhos e camundongos imunizados com o extrato salino apresentaram diminuição da carga parasitária oriunda da infecção posterior com cercárias do S. mansoni, em relação aos controles. Os percentuais de proteção foram de 88.6% e 54% para os animais vacinados (coelhos e camundongos respectivamente)
Studies were carried out with a polyribosomal fraction isolated from Trypanosoma cruzi Y epimastigotes, with the intention to determine both its immunogenic activity and the degree of protection it could induce against experimental T. cruzi infection. This fraction was assayed in four groups of mice by using different schedules of vaccination and varying the dose, intervals, and route of administration. Seven days after the last dose, the animals were sacrificed for immunological studies or subjected to challenge with T. cruzi trypomastigotes. The results obtained in all schedules showed that our polyribosomal fraction only induced a weak antibody response, but was capable of evoking an expressive cellular response. It was also shown that this fraction has the capacity of inducing a high degree of protection against T. cruzi infection, as determined by the decrease of parasitemia and the prolonged survival time of immunized animals.
Viable metacyclic forms of T. cruzi, Y strain, treated with an adequate dose of actinomycin D (50 micrograms Act-D/ml/10(7) parasites/ml for 72 h at 28 degrees C) showed the following properties: 1) they lost their ability to replicate in culture medium, in blood and in tissues of normal mice and were no longer able to incorporate tritiated thymidine; 2) they could not penetrate into Vero cells and could not replicate inside normal macrophages; 3) they retained their immunogenicity and the ability to protect mice against a virulent infection; 4) they did not induce histological lesions as described in chronic experimental Chagas' disease.
The subcutaneous (s.c.) vaccination of DBA/2 mice with 4 weekly doses of 3 x 10(7) living metacyclic forms of T. cruzi, Y strain, obtained from culture in axenic medium and treated for 24 h with actinomycin-D (50 micrograms/10(7) parasites), a drug that promotes an irreversible blockade of the parasite replication, do not induce any detectable degree of humoral and cellular immunosuppression as assessed by a) the production of anti-SRBC antibodies, b) the permanence of delayed cutaneous reaction to T. cruzi antigen, to PPD and DNCB and c) the degree of blastogenic transformation of spleen lymphocytes in the presence of the specific antigen.
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