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Zorgi NE. Humoral immunity in BALB/c and C57Bl/6j mice immunized with irradiated tachyzoite of Toxoplasma gondii. [Master Thesis (Parasitology)]. Sao Paulo: Instituto de Ciencias Biomedicas da Universidade Sao Paulo; 2010. Toxoplasmosis is a highly disseminated infection, affecting one fifth of the human population. T.gondii presented a complex life cycle involving cats and warm blood hosts, including man. Component vaccines are usually poor immunogens and living attenuated vaccines cause chronic disease in the host. 255Gy irradiated tachyzoites do not cause infection in the host and induce the same immunity as natural infection. Challenge of immunized animals resulted in partial protection, without studies on immunological memory. Here, we studied in vitro production of specific IgG by cells from spleen and bone marrow of isogenic mice immunized parenterally or per os with irradiated tachyzoites, comparing with chronically infected animals. Spleen and bone marrow cells were isolated from those animals and, after in vitro growth, the production specific antibodies were measured by ELISA, total or high affinity, after chaotrope treatment. Protection was measured by numbers of brain cysts, one month after challenge with three different strains. All immunized models presented specific IgG in their serum. After four days in vitro growth, bone marrow and spleen cells clearly produced specific IgG similar as chronic infection in parenterally immunized mice, higher than per os immunized animals. Bone marrow contains larger numbers of specific IgG producing cells than spleen. Most of the specific antibodies were of high affinity in all immunizations, diverse from serum which presented low proportion of high avidity IgG. Preliminary results show also presence of specific IgA antibodies in stools. All immunizations induce partial protection against challenge, which was more evident in highly pathogenic ME-49 strain. The protection evidenced was proportional both to the amount of IgG antibodies in serum or produced by bone marrow cells, after longer culture periods. Our data clearly shows that the amount of high affinity antibodies is related to specific memory cell survival, especially in bone marrow. Immunization with irradiated tachyzoites is a good vaccination model for experimental toxoplasmosis, regardless of challenging parasite strain.

Imunidade humoral de camundongos BALB/c e C57BI/6j imunizados com taquizoítos de Toxoplasma gondii irradiados