ABSTRACT
1. Louvain rats (IgK-1a) were immunized with horse IgG(T). To generate mAb to IgG(T), popliteal lymph node cells taken from the immunized animals were fused to a non-secreting LOU/C immunocytoma (IR983F). The hybridomas were cultured in HAT -containing medium and cloned under limiting dilution conditions. Supernatants from the growing hybrids were screened by ELISE using plates coated with horse IgG(T) or IgGa+b+c. 2. The anti-IgG(T) mAb obtained was named LO-HoGT-1 (LOU anti-horse IgG(T)). It is an IgG2a rat antibody whose light chain allotype is IgK-1a, and with an affinity constant of 2.9 x 1010 M-1. 3. Ascites was isnduced in LOU (IgK-1b) rats by injecting the hybridoma cells and incomplete Freund's adjuvant ip. To obtain purified mAb, ascitic fluid was applied to a Sepharose anti-rat LOU IgK-1 a chain column. 4. The purified mAb was then coupled to Sepharose. Immunoelectrophoretically pure IgG(T) was obtained by passage of horse serum through this column. The entire procedure took less than 30 min and resulted in a highly purified IgG(T)
Subject(s)
Animals , Female , Male , Rats , Antibodies, Monoclonal/isolation & purification , Horses/immunology , Immunization , Immunoglobulin G/isolation & purification , Antibodies, Monoclonal/immunology , Cell Line , Chromatography, Affinity , Chromatography, Agarose , Enzyme-Linked Immunosorbent Assay , Hybridomas/immunology , Immunoelectrophoresis , Immunoglobulin G/immunologyABSTRACT
Las ratas Fischer (F344) machos infectadas experimentalmente con diferentes cantidades de tripomastigotes sanguineos de la cepa Tehuantepec de Trypanosoma cruzi desarrollan un aumento progresivo de la parasitemia, seguida por la muerte de los animales en la fase aguda de la infeccion.Las curvas de parasitemia muestran un patron comun y la muerte de los animales ocurre en periodos definidos. En esta cepa de ratas fue posible inducir una proteccion contra la enfermedad aguda despues de inocularlas con epimastigotes vivos, de cultivo. No se produjeron parasitemias al examen directo, no obstante una infeccion subyacente fue detectada tras cultivar celulas del bazo de animales protegidos. En el suero de todos los animales se demostro la presencia de anticuerpos anti-T. cruzi y altos niveles de IgG2a