ABSTRACT
Monoclonal antibodies against tetanus toxin were produced to obtain highly specific antisera. Ten hybridoma cell lines producing monoclonal antibodies were derived from the fusion of rat myeloma cells and spleen cells from rats immunized with tetanus toxoid. Eight produced monoclonal antibodies specific for determinants on toxin and toxoid, whereas two were specific only for determinants on the toxoid. The antibodies produced by hybridomas were characterized by determination of the class of light and heavy chain components, epitope specificity, toxin neutralization, and subunit specificity. All of the antibodies contained kappa light chain, eight contained the gamma 1 heavy chain, and the remaining two contained the gamma 2a heavy chain. Five distinct epitopes were indicated by competition assay of paired monoclonal antibodies, and 4 of the 10 monoclonal antibodies neutralized the in vivo activity of tetanus toxin. The four neutralizing monoclonal antibodies and one other were specific for the C fragment of the heavy chain of the toxin molecule.
Subject(s)
Antibodies, Monoclonal/immunology , Tetanus Toxin/immunology , Animals , Electrophoresis, Polyacrylamide Gel , Epitopes/analysis , Hybridomas/analysis , Molecular Weight , RatsABSTRACT
Four soluble antigens of Mycobacterium leprae have been identified using 12 murine monoclonal antibodies. Their specificity, taxonomic distribution and molecular nature were analysed by radioimmunoassays and by immunoblotting from polyacrylamide electrophoresis gels. A protein antigen MY1 (12K) reacted with one antibody (ML06) without demonstrable cross-reactivity for any of the other 20 tested species of mycobacteria. Another four antibodies which identified antigen MY2 revealed only a marginal degree of cross-reactivity with three other species of mycobacteria. Two antigens shared by several other mycobacteria species were: MY3 represented by five protein bands (35-70K) and a subtilisin resistant molecule MY4 (40-50K) with two distinct determinants and presumably of polysaccharide nature. The described monoclonal antibodies may represent novel valuable diagnostic reagents as well as tools for the purification of antigens which could be explored towards prophylactic or therapeutic immunization against leprosy.
Subject(s)
Antigens, Bacterial/analysis , Epitopes/analysis , Mycobacterium leprae/immunology , Antibodies, Monoclonal/immunology , Antigens, Bacterial/immunology , Binding, Competitive , Cross Reactions , Electrophoresis, Polyacrylamide Gel , Epitopes/immunology , Molecular Weight , Mycobacterium/immunology , Radioimmunoassay , Species SpecificityABSTRACT
Isomaltohexaose flavazole coupled to chicken gamma-globulin (IM6-CGG) induced T cell-dependent anti-alpha(1 leads to 6) dextran-specific IgM and IgG responses in CBA, BALB/c and A strain mice. The IgG responses were of restricted heterogeneity as judged by isoelectric focusing, and belonged mostly to the IgG1 subclass with a minor IgG3 component in the case of BALB/c and CBA mice. All four subclasses of IgG were produced in A strain mice. In contrast, native dextran B 512 induces exclusively T cell-independent IgM responses of the same specificity for all 3 strains. All BALB/c mice immunized with different doses of IM6-CGG either in Freund's adjuvant or with Al(OH)3 plus Bordetella pertussis showed the same spectrotypes by isoelectric focusing. Sera obtained at different times after immunization of BALB/c mice failed to show spectrotype variations. Late, but not early, bleedings from CBA mice showed a tendency towards more uniform isoelectric focusing patterns.
