No abstract

Peripheral blood leukocytes from individuals immunized with tetanus toxoid can be stimulated by pokeweed mitogen to produce IgG anti-tetanus toxoid antibody (IgG-Tet) in vitro. Previous studies have shown that treatment of these cells with tetanus toxoid or anti-human IgG reagents can inhibit this in vitro antibody synthesis. We have examined the four IgG subclasses on the surface of B cells for their relative contributions in the anti-IgG antibody-induced inhibition of IgG-Tet production. With all donors, the inclusion of anti-IgG1, but not anti-IgG2, -IgG3, or -IgG4, antiserum resulted in the in vitro inhibition of IgG-Tet synthesis. The magnitude of this inhibition was similar to that induced by treatment of the B cells with tetanus toxoid antigen. When the supernatants from normal in vitro cultures were assayed for IgG-Tet of the various IgG subclasses, it was observed that the IgG-Tet were almost exclusively IgG1. Similar results were obtained when serum IgG-Tet were measured. Thus, IgG1 appears to be the major subclass for (1) the in vivo-produced IgG-Tet, (2) the in vitro-produced IgG-Tet, and (3) the membrane receptor which can selectively convey an inhibitory signal to the IgG-Tet B cell.

SUMMARYWe investigated the role of apoptosis in the differentiation failure of B cells from a selected subpopulation of patients with CVID delineated by B cell surface marker analysis, in vitro IgE response, and molecular markers of B cell VH gene repertoire. These patients had altered display of B cell surface molecules that play a role in apoptosis. The patients' B cells had a 4 5-250-fold increase in CD95 (Apo-l, fas) expression and increased CD95 display on their T cells. CD38, a molecule important in preventing germinal centre B cell apoptosis, was reduced on the patients' B cells. The expression of this molecule was inducible on the CVID lymphocytes with retinoic acid. Increased spontaneous apoptosis in vitro was observed with the patients' B (23%) and T cells (10%) compared with normal cells (13% and 3%, respectively). Stimulation in vitro with IL-4 and CD40 rescued the B cells from apoptosis and allowed for their differentiation. However, IL-4 plus aCD40-driven immunoglobulin production was not quantitatively or qualitatively normal. Failure to overcome apoptosis, a normal step in germinal centre B cell development, may be involved in the lack of differentiation seen in this subset of CVID patients.Keywords common variable immunodeficiency human humoral immunodeficiency antibody deficiency B cell development immunoglobulin production

In this study we have defined the parameters needed for the optimum detection of anti-Fab antibodies in the serum of patients with rheumatoid arthritis. We have found that the majority of the anti-Fab antibodies are of the IgG3 and IgG4 subclasses which were not optimally detected using polyclonal heterologous anti-human IgG antisera; subclass-specific antibodies instead were needed. Additionally we determined that dissociation of circulating immune complexes by dialysis against urea for 3–7 days was also needed for the detection of these antibodies. Lastly we have shown that the dissociated complexes can recombine with their target Fab molecules, and therefore separation of the anti-Fab antibodies from the other immunoglobulins by chromatofocusing may enhance the detection of these antibodies. When the above conditions were fulfilled it was determined that IgG anti-Fab antibodies could be detected in rheumatoid arthritis and normal sera and that acidic IgG3 and IgG4 subclasses predominated. However, IgG3 levels were 10.5-fold higher in rheumatoid arthritis sera (p <0.05) and IgG4 levels 5-fold higher (p <0.01) than in normal sera.

Anti-Fab antibodies (aFABA) of restricted clonality and acidic spectrotypes were isolated from the sera of patients with rheumatoid arthritis (RA). These aFABA reacted with multiple populations of pooled human Fab molecules, which had been charge separated by chromatofocusing techniques (CF), indicating that the structures recognized by these aFABA were present on a polyclonal population of Fab molecules. The structures were also widely distributed among the Fab repertoires of normal individuals, as well as individual autologous and heterologous RA patients. Thus, the aFABA did not appear to recognize highly restricted epitope(s), i.e. a private idiotope, limited in its expression to RA individuals. The determinants of the Fab molecules recognized by affinity purified aFABA could be defined by linear and/or conformational structures, depending upon the individual from which the aFABA were isolated. Additionally, some of the affinity purified aFABA also reacted with Fc fragments, suggesting the presence of epibody-like autoantibodies in this population. Lastly, size analysis of the circulating IgG4 aFABA complexes indicated that these autoantibodies were not complexed with intact IgG, but rather with a molecule of 40-60 kDa, further suggesting the potential for these autoantibodies to react with multiple antigens.