ABSTRACT
Crithidia fasciculata is an important trypanosomatid parasite commonly affecting insects and is used extensively as a model for the study of the biochemistry, ultrastructure and organization of the kDNA network of trypanosomatids. The present study describes the evolution of UV-induced morphological changes detectable by transmission electron microscopy in Crithidia fasciculata. Although only rare and minor changes in Kinetoplast DNA were demonstrable 7 h after UV irradiation, alterations of this orgtanelle were present in almost al flagellates observed 24 h and 48 h after irradiation. Other cell structures were apparently undamaged. Ultrastructural changes in kDNA did not correspond to changes in antigenicity of protein bands in western blotting against serum from Chagas' disease patients or in the presence of 3 different lectin receptors on the surface of the parasite
Subject(s)
Antigens/radiation effects , Crithidia/ultrastructure , DNA/radiation effects , Lectins/radiation effects , Ultraviolet Rays , DNA/ultrastructure , Microscopy, Electron , MutagensABSTRACT
The need for a reliable method for the immunological diagnosis of Kala-azar is imperative. Leishmania donovani donovani and L. donovani chagasi culture promastigotes were compared as antigens in a direct agglutination test (DAT) for the diagnosis of visceral leishmaniasis in Brazil. Both antigens were successfully employed for the DAT, Showing 100½ sensitivity and > 98% specificity when used to test sera from Brasilian and African Kala-azar, Chagas'disease, malaria, filaria and symphilis patients, and on sera from Brazilian controls. Cross-reactions were sometimes observed when cutaneous and cucocutaneous leishmaniasis patient sera were teste. The vross-reactions were completely abolished by the addition of 0.78% 2-mercaptoethanol to the serum diluent. These data show data that this improved Dat can be used for the diagnosis of visceral leishmaniasis in Brazil
Subject(s)
Humans , Antigens, Protozoan/analysis , Leishmania donovani/immunology , Leishmaniasis, Visceral/diagnosis , Agglutination TestsABSTRACT
The effects of short ultraviolet (253 mm) radiation on the growth of the trypanosomatid flagellate Crithidia fasciculada were studied in liquid medium and nutrient agar plates. Cell duplication was completely inhibited after exposure of the flagellates to doses equal to or higher than 50 J/m2. The UV-induced lag period was dose-dependent. Survival was reduced to 1% after exposure of the parasites to 200 J/m2. Ultrastructural changes after the lag period were studied by transmission electron microscopy. Changes of the kinetoplast network structure and sometimes of the mitochondrial matrix were observed. The existence of DNA repair mechanisms in this protozoan is discussed
Subject(s)
Animals , Cell Division/radiation effects , Crithidia/radiation effects , DNA Repair/radiation effects , Radiation Dosage , Ultraviolet Rays , Culture Media , Dose-Response Relationship, Radiation , Regression AnalysisABSTRACT
An enzyme-linked lectin assay (ELLA) based on the ELISA assay, using intact formalin-fixed promastigotes to coat poly-L-lysine-treated microtiter plates is described. The assay was used to study the lectin receptors of Leishmania donovani chagasi, L. donovani donovani and L. mexicana amazonensis. ConA, RCA, WGA, and PNA receptors were found in the three parasites. SBA receptors were found to be as frequent as the other receptors in L. donovani chagasi but not in the other two parasites which showed little SBA binding. Trypsin treatment of the two L. donovani subspecies did not remove any of the lectin receptors studied
Subject(s)
Animals , Enzyme-Linked Immunosorbent Assay , Leishmania donovani/immunology , Leishmania mexicana/immunology , Receptors, Mitogen/metabolism , Glycoproteins/metabolism , Trypsin/pharmacologyABSTRACT
L. donovani antigens were analyzed by a direct agglutination test (DAT), by ELISA using intact promastigotes and by the SDS-PAGE immunoperoxidase technique (SGIP). Sera of Chagas' disease patients cron-reacted in the ELISA and SGIP but not in the DAT. Trypsin treatment of the parasites removed concanavalin A-binding sites but not epitopes for antibodies present in Chagas' disease and in leishmaniasis sera, as seen in the SGIP. Eight bands were revealed after incubation of the gel sections with Kala-azar or Chagas' disease sera, three of which were common to both sera. The major antigenic component recognized by leishmaniasis sera was a glycoprotein of 57 KD, and the major cross-reacting protein recognized by Chagas' disease sera was a glycoprotein doublet of 71.5-68KD