RESUMEN
A series of hybridomas producing monoclonal antibodies to Crimean hemorrhagic fever virus was obtained and their cultural properties were characterized. HEMA-12 and HEMA-24 secreted monoclonal IgG2b antibodies, HEMA-101 secreted monoclonal IgG1 antibodies, HEMA-31, HEMA-9 and HEMA-11 secreted monoclonal IgG2 antibodies. According to the results of the indirect immunofluorescence test, the titer of specific immunoglobulins in the culture fluid was 1:16-1:32, but sometimes reached 1:64-1:128. The titer of antibodies in ascitic fluid amounted to dilutions of 10(4)-10(5). Hybridomas were cloned by the method of ultimate dilutions. The injection of 5-15 million HEMA cells into the abdominal cavity of BALB/c mice induced the formation of ascitic tumors in the animals within 7-11 days and the accumulation of ascitic fluid in a volume of 1-4 ml. Hybridomas, found to be capable of passage from mouse to mouse, underwent 5-16 passages during the term of observation.
Asunto(s)
Anticuerpos Monoclonales/biosíntesis , Fiebre Hemorrágica de Crimea/inmunología , Hibridomas/inmunología , Animales , Inmunoglobulinas/biosíntesis , Ratones , Ratones Endogámicos BALB CRESUMEN
Twelve lines of hybridomas secreting monoclonal antibodies (MCA) have been generated by fusion of spleen lymphocytes of BALB/c mice immunized with crude Crimean hemorrhagic fever virus (CHF). The hybridomas multiply actively in vitro (over 50 passages) and as ascitic tumors in the abdominal cavity of BALB/c mice. MCA were characterized by indirect immunofluorescence (IF), complement fixation (CF), biological neutralization test (NT), agar gel diffuse precipitation tests, and by the type of immunoglobulins. In the indirect IF, all kinds of MCA reacted with CHF virus, in CF only GEMA 12 and GEMA 31 which also precipitated the virus antigen in AGDPT. The CF-positive MCA belonged to the IgG2a and IgG2b subclasses, but 2 more MCA species of the same immunoglobulin type did not react in CF. No neutralizing properties have been found with MCA. All MCA reacted similarly in indirect IF and CF both with CHF and Congo viruses. Among other members of the CHF-Congo antigenic group, only GEMA 4 MCA interacted with Hazara virus. MCA generated in this study are intended for use in identification of CHF and Congo viruses in diagnostic tests.