Functional differentiation of B lymphocytes in congenital agammaglobulinemia. II. Immunochemical analysis of the in vitro primary immune response.
Cultures of peripheral blood lymphocytes (PBL) in which specific hemolytic plaque-forming cells (HcPFC) had been induced were labeled with 14C-amino acids. Antigen-specific products in the culture supernatants were characterized by using indirect immune precipitation in conjunction with specific immunoabsorbents and/or gel filtration followed by SDS-polyacrylamide gel electrophoresis. After 5 days of culture with antigen (sheep red blood cells or ovalbumin) newly synthesized IgM and specific IgM antibody were demonstrated in culture supernatants from normal donors and from four out of five patients with congenital agammaglobulinemia (cAgamma). Secreted products bound specifically to antigen and pretreatment of labeled supernatants with anti-mu and anti-L chain antisera, but not with anti-gamma antiserum, prevented binding. Typical mu- and L chains constituted only a proportion of the anigen-binding peptides recognized by the anti-mu reagents. Induction of IgM antibody synthesis was dependent on the presence of antigen and was correlated with the generation of HcPFC. No major differences between the antigen-induced products of cAgamma and normal PBL were observed. These findings suggest that in the absence of terminal B cell differentiation in vivo, certain patients with cAgamma possess precursor cells that can respond to antigen in vitro with the synthesis of specific humoral products, including IgM antibody.
D000042:Absorption; D000361:Agammaglobulinemia / Q000151:congenital* / Q000276:immunology; D000917:Antibody Formation; D001402:B-Lymphocytes / Q000166:cytology*; D001665:Binding Sites; D002250:Carbon Radioisotopes; D002454:Cell Differentiation; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D004912:Erythrocytes / Q000276:immunology; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000032:analysis*; D007147:Immunoglobulin Light Chains / Q000032:analysis*; D007075:Immunoglobulin M / Q000032:analysis / Q000096:biosynthesis; D007148:Immunoglobulin mu-Chains / Q000032:analysis*
D002250:Carbon Radioisotopes; D000939:Epitopes; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007075:Immunoglobulin M; D007148:Immunoglobulin mu-Chains