FUNCTIONAL DIFFERENTIATION OF LYMPHOCYTE-B IN CONGENITAL AGAMMAGLOBULINEMIA .2. IMMUNOCHEMICAL ANALYSIS OF INVITRO PRIMARY IMMUNE-RESPONSE

  • 1 January 1977
    • journal article
    • research article
    • Vol. 119  (6) , 1965-1972
Abstract
Cultures of peripheral blood lymphocytes (PBL) in which specific hemolytic plaque-forming cells (HcPFC) were induced with 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[sodium dodecyl sulfate]-polyacrylamide gel electrophoresis. After 5 days of culture with antigen (sheep red blood cells or ovalbumin) newly synthesized Ig[immunoglobulin]M and specific IgM antibody were demonstrated in culture supernatants from normal donors and from 4 out of 5 patients with congenital agammaglobulinemia (cA.gamma.). 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 antigen-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 cA.gamma. and normal PBL were observed. In the absence of terminal B [bone marrow-derived] cell differentiation in vivo, certain patients with cA.gamma. possess precursor cells that can respond to antigen in vitro with the synthesis of specific humoral products, including IgM antibody.

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