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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.

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.