Different patterns of inositol polyphosphate production are seen in B lymphocytes after cross-linking of sIg by anti-Ig antibody or by a multivalent anti-Ig antibody dextran conjugate

J Immunol. 1989 Sep 1;143(5):1414-21.

Abstract

Anti-delta antibody conjugated to 2 x 10(6) m.w. dextran (dex) stimulates B lymphocyte proliferation at 10,000-fold lower concentrations than that required by the unconjugated antibody. Dex conjugated antibody also stimulates a greater and more sustained increase in intracellular ionized calcium [( Ca2+]i) than does the unconjugated anti-Ig antibody. Inasmuch as inositol phosphate metabolites have been linked to rises in [Ca2+]i, we analyzed by FPLC the relative amounts of the inositol polyphosphates (IP) in these cells. Anti-Ig-dextran induced a threefold greater increase in total IP than did the unconjugated anti-Ig. Furthermore, in cells stimulated by unconjugated anti-Ig there was a transient induction of I(1,4,5)P3 followed by a rapid accumulation of the I(1,3,4)P3 isomer with little accumulation of I(1,4)P2, whereas in anti-Ig-dex-stimulated cells there was prolonged elevation of I(1,4,5)P3 with more accumulation of I(1,4)P2. In addition, levels of I(1,3,4,5)P4 were maintained over a longer period of time in B cells stimulated by anti-Ig-dex than in those stimulated by unconjugated anti-Ig. The enhanced ratio of I(1,4,5)P3/I(1,3,4)P3 was also seen when suboptimal concentrations of anti-Ig-dex were used which stimulated a level of total inositol phosphate that was similar to that stimulated by the unconjugated anti-Ig. The possibility that the greater stimulation of increased [Ca2+] by anti-Ig-dex than by unconjugated anti-Ig was a predominant factor in influencing the metabolic pathway of I(1,4,5)P3 was excluded. These results show that 1) stimulation of increases in the various IP isomers occurs in anti-Ig stimulated normal B cells as has been shown in B cell lines and 2) that signal transduction and consequent PIP2 hydrolysis that is stimulated by Ag-mediated cross-linking of sIg is strongly influenced by the extent and type of cross-linking that is induced.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Anti-Idiotypic / pharmacology*
  • B-Lymphocytes / metabolism*
  • Calcium / metabolism
  • Cross-Linking Reagents / pharmacology*
  • Cytosol / metabolism
  • Dextrans / pharmacology*
  • Dose-Response Relationship, Immunologic
  • Female
  • Hydrolysis
  • Immunoglobulin D / immunology*
  • Immunoglobulin D / pharmacology
  • Inositol Phosphates / biosynthesis*
  • Mice
  • Mice, Inbred DBA
  • Mice, Nude
  • Phosphatidylinositol 4,5-Diphosphate
  • Phosphatidylinositols / metabolism
  • Receptors, Antigen, B-Cell / metabolism*
  • Receptors, Antigen, B-Cell / pharmacology
  • Sugar Phosphates / biosynthesis*

Substances

  • Antibodies, Anti-Idiotypic
  • Cross-Linking Reagents
  • Dextrans
  • Immunoglobulin D
  • Inositol Phosphates
  • Phosphatidylinositol 4,5-Diphosphate
  • Phosphatidylinositols
  • Receptors, Antigen, B-Cell
  • Sugar Phosphates
  • anti-IgD
  • Calcium