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One percent of human circulating B lymphocytes are capable of producing the
natural anti-Gal antibody
U Galili, F Anaraki, A Thall, C Hill-Black and M Radic
Department of Microbiology and Immunology, Medical College of Pennsylvania,
Philadelphia 19129.
The natural anti-Gal antibody constitutes 1% of circulating IgG in humans
and interacts specifically with the carbohydrate epitope Gal alpha 1-3Gal
beta 1-4GlcNAc-R (the alpha-galactosyl epitope). In view of the unusually
large amounts of this antibody in the serum, it was of interest to
determine the proportion of circulating B lymphocytes capable of
synthesizing anti-Gal. For this purpose, blood B lymphocytes were incubated
with Epstein-Barr virus (EBV) and plated in microtiter wells. Proliferation
of the EBV transformed B lymphocytes was readily visible after 3 weeks of
incubation. The supernatants from wells containing proliferating B-lymphoid
clones were assayed for anti-Gal by an agglutination assay with rabbit red
blood cells and the specificity of the agglutinating antibodies was further
confirmed by their interaction with synthetic oligosaccharides and by
enzyme-linked immunosorbent assay with glycoproteins. Approximately 5% of
the wells contained anti-Gal antibodies. Limiting dilution studies and IgH
gene rearrangement patterns suggested that each well contained an average
of five proliferating B-lymphoid clones. Thus, it is concluded that
approximately 1% of circulating B lymphocytes are capable of producing
anti-Gal. The proportion of anti-Gal--producing lymphoid clones exceeds by
fourfold that of clones producing anti-blood group A or anti-blood group B
antibodies. Individual anti-Gal clones display fine variations in their
combining site, as indicated by their differential interaction with
alpha-galactosyl epitopes on glycolipids and on N-linked carbohydrate
chains of glycoproteins. The high frequency of precursor B lymphocytes
capable of producing anti-Gal, found in every individual and the restricted
specificity of this antibody to alpha-galactosyl epitopes, potentially
makes anti-Gal--producing lymphocytes an effective system for studying
human Ig genes involved in the natural immune response to structurally
defined haptens.
Volume 82,
Issue 8,
pp. 2485-2493,
10/15/1993
Copyright © 1993 by The American Society of Hematology
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