SUMMARYA comparativ e stud y o f th e antigeni c profil e o f bloodstrea m an d cel l cultur e derived trypomastigote s showe d many difference s i n thei r components. Using mouse anti-T. cruz i antibodie s th e difference s wer e locate d mostl y i n th e 12 0 kDa band , whereas usin g chagasi c patien t ser a th e difference s wer e locate d i n th e 8 5 an d 5 2 kDa bands . These finding s migh t explai n know n physiologica l difference s betwee n trypomatigotes obtaine d fro m cel l cultur e an d fro m infecte d blood . A brie f repor t of thi s wor k ha s already bee n published 9 .
SUMMARYSera of Chaga's disease patients containing anti-T. cruzi lytic antibodies were submitted to affinity chromatography using Sepharose 4B conjugated with antigen extracted from epimastigote or trypomastigote forms of the parasite. Epimastigotes were obtained from culture at the exponential growth phase and the trypomastigotes from blood of infected and immunosuppressed mice. Antigen of both parasite forms was obtained by sonication of the parasites followed by cemrifugation. Both antigens were then conjugated to activated Sepharose 4B. Affinity chromatography was performed by passing sera from chagasic patients through an immunoadsorbent column containing either epimastigote or trypomasugote antigens. Antibodies bound to the column were eluted with cold 0,2 M glycine buffer pH 2,8. The eluted antibodies were analysed regarding their isotype and lytic activity. The results showed that anti-T. cruzi lytic antibodies present in sera from chagasic patients are mainly located in the IgG isotype and recognize epitopes present in both trypomastigote and epimastigote forms. A brief report of this work has already been published 12 .
Bloodstream trypomastigotes were isolated from blood of A/Sn mice 7 d after infection with 10(5) Trypanosoma cruzi Y strain. Red blood cells were removed by centrifugation and hypotonic shock and platelets and leucocytes by passage through a carboxy methyl cellulose column. Binding of trypomastigotes to the resin was prevented by including 10% normal mouse serum in the eluting buffer. In such conditions, more than 90% of the parasites applied to the column were recovered, free of white blood cells and platelets. A comparative study of the pre- and post-separation trypomastigotes showed that both had the same infecting capacity, ability to evade destruction by the complement system, and antigenic profile.
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