[Show abstract][Hide abstract] ABSTRACT: SKF525A, an inhibitor and inducer of cytochrome P450, was tested on different developmental stages of Trypanosoma cruzi. Growth, motility and structure of epimastigotes, motility and infectivity of trypomastigotes, and infectivity of trypomastigotes to Vero cells in culture were abolished by the drug at 10-100 microM concentrations. When blood from infected mice was treated with the drug, and used to infect 8 day-old mice, no parasites were observed at 0.6-1 mM, and all animals survived. Blood cell morphology was well preserved, and the sleeping time of pentobarbital-treated mice inoculated with the same amount of drug was not increased. The present results suggest that SKF525A or other related inhibitors of cytochrome P450 coned be tested as an additive for blood sterilization in blood banks.
[Show abstract][Hide abstract] ABSTRACT: In this work it was shown that the infectivity of trypomastigote forms of Trypanosoma cruzi was affected upon the interaction with the Monoclonal Antibody (McAb) 2E9, which was raised against a glycoconjugated fraction of membranes of epimastigotes (Tulahuen strain). Characterization of the epitope recognized by this McAb, as well as its effects on complement mediated lysis and host cell invasion are reported. Immunocytochemical analysis showed that the McAb was reactive with two macromolecules (41-58 kDa) present on Trypanosoma cruzi epimastigotes (Tulahuen and Y strain), while it recognized several trypomastigotes macromolecules, showing a more intense reactivity with a band of 80 kDa. By indirect immunofluorescence, it was found there were subpopulations of blood and tissue culture derived trypomastigotes which attach the antibody to varying degrees. Studies using chemical or enzymatically treated antigens suggested that the McAb 2E9 was directed against carbohydrate epitopes, which were identified as being--galactosyl residues. In addition, preliminary results are shown, suggesting that the epitope recognized by the McAb 2E9 is involved in adhesion/or internalization of trypomastigotes.
No preview · Article · Jan 1997 · Revista latinoamericana de microbiología
[Show abstract][Hide abstract] ABSTRACT: Subcellular fractions of T. cruzi epimastigotes (Epi) were studied for their capability to induce protective or aggressive effects in animals. The flagellar fraction (F) showed the best immunoprotective properties without tissular aggression. Monoclonal antibodies were raised against F. Two of them, FCH-F8-1 and 4, were able to neutralize the infectivity of bloodstream forms, to mediate lysis by complement of cell culture derived[trypomastigotes (Tripo) and to recognize the surface of Tripo and Epi. FCH-F8-1 reacted with a 85 kDa protein from Tripo (assayed by immunoprecipitation) and with peptides of 43 kDa on Epi and Tripo (tested by immunoblotting). FCH-F8-4 recognized several proteins ranging from 50 to 150 kDa on Epi and two molecules of 15 and 48 kDa on Tripo. Mice immunized with antigens purified by affinity chromatography by using FCH-F8-4 were protected against the infection. Several recombinant clones were detected on a cDNA lambda gt11 expression library constructed from T. cruzi Epi (Tulahuén strain): three with FCH-F8-4 and two with FCH-F8-1. One clone recognized by each monoclonal antibody was studied gamma (FCH-F8-1) 1 and gamma (FCH-F8-4) 1. Both inserts were of 150 base pairs each; they detected a 3.5 and 5.0 kilobases Epi mRNA, respectively. Both inserts were sequenced, and the amino acid sequences were inferred. gamma (FCH-F8-4) 1 codified for a 19 aa peptide, PAFLGCSSRFSGSFSGVEP, and gamma (FCH-F8-1) 1 for a 29 aa peptide EFLERGRISCORHSYTSYTSCSDEHNVTPFC. The whole 19 aa peptide was synthesized. This peptide (SP4) inhibited the ELISA reactivity against the parasite of chronically infected and F immunized mouse sera.(ABSTRACT TRUNCATED AT 250 WORDS)
[Show abstract][Hide abstract] ABSTRACT: Trypanosoma cruzi, the causative agent of Chagas' disease, is widely spread in Central and South America. The present report describes three monoclonal antibodies (mAbs) directed against the flagellar fraction of epimastigotes (Ffe) of the parasite, Tulahuén strain. The three mAbs were of IgG1 isotype. Indirect immunofluorescence assays revealed that the three mAbs bind to epimastigotes, the FCH-F8-1 and -3 bind to blood trypomastigotes (BT) and FCH-F8-1 is the only one that binds to amastigotes. Three different proteins of the parasite were recognized by the mAbs in immunoprecipitation assays: an 85 kDa of BT with the FCH-F8-1 mAb, a 40 kDa of Ffe with the FCH-F8-2 mAb, and a 90 kDa of Ffe and of BT with the FCH-F8-3 mAb. Positive complement mediated lysis (CML) of BT and metacyclic forms, obtained from the insect vector feces, were shown by the FCH-F8-1, while FCH-F8-3 only showed CML against the metacyclic trypomastigotes. FCH-F8-2 did not mediate any CML activity. Passive transference of the mAbs to BALB/c mice conferred protection, in terms of survival, ranging from 50 (FCH-F8-2 and -3) to 80% (FCH-F8-1) against a challenge with 1 X 10(3) BT. These results suggest that FCH-F8-1 may be a useful tool to purify proteins in order to investigate their role in immunoprotection experiments.
No preview · Article · Nov 1986 · Immunology Letters
[Show abstract][Hide abstract] ABSTRACT: Mice were immunized with subcellular fractions obtained by differential centrifugation from epimastigotes of Trypanosoma cruzi (Tulahuén strain). In a chronic model of Chagas' disease, they were challenged each with 25 bloodstream trypomastigotes. Non-immunized, non-challenged and non-immunized challenged animals were kept as controls. Among the challenged mice, those immunized with 105,000 g pellet (Mc) and 105,000 g supernatant (Cs) fractions presented positive xenodiagnosis, myocarditis and myositis similar to those shown by non-immunized challenged controls. The fractions enriched in flagella, the 5000 g pellet (P5) and the flagellar fraction (F) resulted in fewer animals with positive xenodiagnosis and in hosts partially protected from the development of myocarditis. In the absence of infection, Mc and Cs induced an intense myocarditis while F induced mild lesions similar to those found in the controls. P5 caused a myocarditis intermediate between that elicited by Mc and that in the controls. 50% of the animals immunized with Cs presented pathological electrocardiograms in the absence of challenge. The animals immunized with F and P5 and challenged were protected against the development of pathological electrocardiograms, whereas those immunized with Mc and Cs behaved like the non-immunized controls. The immunized, non-challenged animals presented anti-T. cruzi IgG antibodies, with titres which were lower than those shown by the immunized and challenged mice.--The results show the possibility of obtaining tissue lesions with antigenic preparations of T. cruzi in the absence of infection, and suggest that the mechanisms involved in the generation of myocarditis and electrocardiographic alterations are probably different, since these pathologies can be elicited by different subcellular fractions. Among the antigenic components of the parasite, the flagellar fraction gave the best immunoprotective properties, with little or no immunoaggressive effects.
[Show abstract][Hide abstract] ABSTRACT: Immunization of mice with subcellular fractions of C. fasciculata led to myocarditis and electrocardiographic alterations similar to those induced by immunization with T. cruzi, the etiological agent of Chagas' disease, suggesting the presence of similar cardiotoxic antigens in both trypanosomatid flagellates.