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, Marko Radic

Research output: Contribution to journalArticle

173 Citations (Scopus)

Abstract

The natural anti-Gal antibody constitutes 1 % of circulating IgG in humans and interacts specifically with the carbohydrate epitope Galα1-3Galβ1- 4GlcNAc-R (the α-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 α-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 α-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.

Original languageEnglish (US)
Pages (from-to)2485-2493
Number of pages9
JournalBlood
Volume82
Issue number8
StatePublished - 1993
Externally publishedYes

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Lymphocytes
Anti-Idiotypic Antibodies
B-Lymphocytes
Clone Cells
Epitopes
Antibodies
Antibody Specificity
Blood Group Antigens
Human Herpesvirus 4
Glycoproteins
Viruses
Carbohydrates
Assays
Blood
Genes
Immunoglobulin Genes
B-Lymphoid Precursor Cells
Gene Rearrangement
Haptens
Glycolipids

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Immunology
  • Medicine(all)
  • Hematology
  • Cell Biology

Cite this

Galili, U., Anaraki, F., Thall, A., Hill-Black, C., & Radic, M. (1993). One percent of human circulating B lymphocytes are capable of producing the natural anti-Gal antibody. Blood, 82(8), 2485-2493.

One percent of human circulating B lymphocytes are capable of producing the natural anti-Gal antibody. / Galili, U.; Anaraki, F.; Thall, A.; Hill-Black, C.; Radic, Marko.

In: Blood, Vol. 82, No. 8, 1993, p. 2485-2493.

Research output: Contribution to journalArticle

Galili, U, Anaraki, F, Thall, A, Hill-Black, C & Radic, M 1993, 'One percent of human circulating B lymphocytes are capable of producing the natural anti-Gal antibody', Blood, vol. 82, no. 8, pp. 2485-2493.
Galili, U. ; Anaraki, F. ; Thall, A. ; Hill-Black, C. ; Radic, Marko. / One percent of human circulating B lymphocytes are capable of producing the natural anti-Gal antibody. In: Blood. 1993 ; Vol. 82, No. 8. pp. 2485-2493.
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abstract = "The natural anti-Gal antibody constitutes 1 {\%} of circulating IgG in humans and interacts specifically with the carbohydrate epitope Galα1-3Galβ1- 4GlcNAc-R (the α-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 α-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 α-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.",
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