ABSTRACT
Lachesis muta snake venom induced aggregation of bromelain sensitized human erythrocytes at a concentration of 1 mg/ml. The hemagglutinating protein was purified by DEAE-Sephadex A-50 column chromatography. Polyacrylamide gel electrophoresis revealed at least three bands, whereas SDS electrophoresis in the presence of 2-mercaptoethanol showed a single one. Isoelectric focusing revealed hemagglutinating activity in the range of pH 3-8. The maximum peak (mutina) at pH 5.5. This fraction was active in agglutinating human RBC of types A, B, O Rh (+) and B, O Rh (-). One mM EDTA and 1 mM Ca++ did not alter the agglutinating time significantly. Lactose and inositol inhibited the agglutination of A, B, O Rh (+) and B, O Rh (-) human RBC. The present study showed the non specificity of the hemagglutinating activity of mutina. It was also shown that mutina is a non-mitogenic protein.
Subject(s)
Crotalid Venoms/analysis , Lectins/isolation & purification , Agglutination , Animals , Chemical Fractionation , Erythrocytes/immunology , Hemagglutination Tests , Platelet AggregationABSTRACT
Four-days old epimastigote culture forms of different stocks of Trypanosoma cruzi from Colombia, Costa Rica and Mexico as well of Colombian stocks of T. rangeli were tested with 27 lectins and the complement lysis test. While the stocks of T. cruzi and T. rangeli showed common agglutination reactions with the lectins of Canavalia ensiformis and Pisum sativum, only the stocks of T. cruzi were additionally agglutinated by Ricinus communis-120, Glycine maxima, Helix pomatia, Axinella polypoides, Bandeiraea simplicifolia, Bauhinia purpurea, Wistaria floribunda, Abrus precatorius, Aaptos papillata II, Limax flavus and Arachis hypogaea. On the basis of lectin typing, the strains of T. cruzi belong to the PNA-type. While the epimastigote culture forms of T. cruzi stocks were lysed by normal fresh human, rat, chicken, rabbit and guinea pig serum, the culture forms of T. rangeli were only lysed by chicken serum. Mouse serum had no lytic effect on either trypanosome species. Incubation of T. cruzi with neuraminidase did not alter the lytic effect of the sera. The membrane-associated N-acetylneuraminic acid on T. cruzi does not protect the cell surface against the activation of the alternative complement pathway. After treatment of T. cruzi with neuraminidase and subsequent incubation in culture medium, the PNA-type was restored. The cells reacted again with the lectins of Arachis hypogaea, Limax flavus, Aaptos papillata II but not with Triticum vulgaris.
Subject(s)
Complement System Proteins/immunology , Lectins/immunology , Trypanosoma/immunology , Animals , Antigens, Protozoan/immunology , Carbohydrates/immunology , Chagas Disease/parasitology , Complement Pathway, Alternative , Humans , In Vitro Techniques , Neuraminidase/pharmacology , Species Specificity , Trypanosoma/classification , Trypanosoma cruzi/immunologyABSTRACT
La accion citolitica de sustancias simples y de agentes biologicos complejos puede ser descrita por ecuaciones tipo Hill. Se propone la medicion del poder hemolitico de venenos de serpientes en terminos de la disminucion de la accion litica en una dilucion 1:10. La ventaja de este procedimiento es que el poder citolitico es una funcion de las propriedades bioquimicas del sistema, mas que de la concentracion de los principios activos. Se muestran los poderes hemoliticos de algunos venenos de serpientes sobre eritrocitos humanos O Rh(+)
