Monoclonal anti-parasite and anti-RBC antibodies produced by stable EBV-transformed B cell lines from malaria patients

Abstract

To produce human monoclonal antibodies associated with infectious disease, peripheral blood lymphocytes (PBL) from patients with Plasmodium falciparum malaria were transformed with EB-virus in vitro. To enrich for malaria-specific B cells, PBL were incubated for 3 days with unsoluble P. falciparum antigen before EBV-transformation. Furthermore, cyclosporin A was added during and after transformation to eliminate T cell suppression of B cell growth. Microcultures were screened for antibodies against blood stage antigens of P. falciparum or of noninfected erythrocytes by ELISA and indirect immunofluorescence. Cultures producing anti-P. falciparum and/or anti-erythrocyte antibodies were developed from the lymphocytes of eight patients, including some individuals with their first infection. Positive cultures were cloned and propagated for several weeks. Seven of 15 clones producing antibody at a stable rate have now been kept in cultures for more than 1 yr. Of six cultures analyzed in detail, all produced IgM antibodies of either K or lambda isotype. Although three clones were monoclonal after one cloning, three were oligoclonal. Of the former, two produced P. falciparum-specific antibodies directed to an antigen associated with the surface of merozoites. One of the oligoclonal cultures produced anti-erythrocyte antibodies, and it was probably reacting with spectrin.