title: Antibody response of rabbit blood lymphocytes in vitro. Kinetics, clone size, and clonotype analysis in response to streptococcal group polysaccharide antigens.
abstract: Peripheral blood lymphocytes (PBL) of rabbits previously hyperimmunized against streptococcal groups A and A-variant antigens were stimulated in vitro by the corresponding vaccines to produce group-specific antibody. This response was dependent on an optimal cell density (2 X 10(6) cells/ml), on the presence of antigen, it was specific and cross-reactive due to a shared rhamnose backbone of the two polysaccharide antigens, and it was highly selective, such that in a 42-55-day culture 1 out of 20 viable cells was a specific PFC. During the exponential increase of the antibody concentration at a constant number of PFC, antibodies were secreted at a rate of 2.4 X 10(4) molecules/s per cell until a plateau level of antibody (40 mug/culture) was reached. The microculture system was used to determine the minimal frequency of group polysaccharide-specific precursor cells in the blood. Independent of the time elapsed since the last immunization this frequency was 1-3 X 10(-5), i.e., in the range of 1-2.8 X 10(2) precursor cells per ml blood. This number was further used together with the clonotype analysis of the culture supernates to calculate the frequencies of precursors of major and minor clonotypes. A hierachy of persisting clonal memory precursor cells was found indicating that clonal dominance is determined by locked-in frequency patterns and therefore it is a phenomenon based on numbers of cells that respond to the antigen.
mesh_terms: D000818:Animals; D000917:Antibody Formation; D002999:Clone Cells; D007106:Immune Sera; D007117:Immunization, Secondary; D007074:Immunoglobulin G / Q000096:biosynthesis*; D007156:Immunologic Memory; D007700:Kinetics; D008214:Lymphocytes / Q000276:immunology*; D011135:Polysaccharides, Bacterial; D011817:Rabbits; D013297:Streptococcus pyogenes / Q000276:immunology*
chemical_list: D007106:Immune Sera; D007074:Immunoglobulin G; D011135:Polysaccharides, Bacterial