RESUMEN
A representative collection was obtained containing 68 monoclonal antibodies (MAB) to Toxoplasma gondii antigens, which was characterized by the binding with the below fractions of tochizoites in the immune-enzyme assay (IEA) and immunoblotting (IB): membrane (MEM), somatic (water-soluble, SOM) and excretory-secretory (ES). Most of MABs were produced to MEM antigens (43), 6 MABs reacted with the somatic fraction, and 3 MABs reacted with both fractions. Two MABs to ES antigen were detected in the latter group. An analysis of MABs in concurrent IEA and IB revealed the immune-dominant proteins of the MEM and SOM fractions of antibodies to T. gondii tochizoites (p30 and p27, respectively). The presence of 2 non-overlapping antigenic determinants was shown for p30. Further research would detect MABs that could be used in the diagnosis of toxoplasmosis.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Antígenos de Protozoos/inmunología , Toxoplasma/inmunología , Animales , Membrana Celular/inmunología , Epítopos , Immunoblotting/métodos , Técnicas para Inmunoenzimas , Ratones , Ratones Endogámicos BALB CRESUMEN
The protein composition and immunochemical properties of Toxoplasma gondii tachyzoites cultured on Vero cells and on mice were studied. Despite the fact that the main components of both preparations were shown to be proteins with molecular weights of 47, 34, 24, and 22 kDa, Toxoplasma-infected human sera antibodies interact mainly with the antigens of 66, 62, 57, 42, 38, 37, 36, 31, and 24 kDa. Comparing efficiency of enzyme immunoassay using the antigens of the tachyzoites obtained in different culture systems showed that the preparation of cultured Vero cells is similar to those of peritoneal exudates from infected mice and may be successfully used for the detection of antitoxoplasma antibodies in the sera of infected subjects.
Asunto(s)
Antígenos de Protozoos/análisis , Toxoplasma/inmunología , Animales , Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/aislamiento & purificación , Líquido Ascítico/parasitología , Chlorocebus aethiops , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunoquímica , Ratones , Peso Molecular , Proteínas Protozoarias/análisis , Proteínas Protozoarias/aislamiento & purificación , Toxoplasma/crecimiento & desarrollo , Toxoplasmosis Animal/parasitología , Células VeroRESUMEN
Chemical-enzymatic synthesis and cloning of a gene for a human anaphylatoxin C5a analog were carried out. Recombinant plasmid pRC5a providing the expression of the synthetic gene in the Escherichia coli cells was obtained. The biological activity of the expression product was demonstrated by the chemotaxis activity test and by the release of myeloperoxidases from rat peritoneal cells that was induced by bacterial cell lysates containing the recombinant protein.