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Mouse B lymphocyte activation of proliferation and cyclic nucleotides at early stages of activation
- A. V. Palladin Institute of Biochemistry, Academy of Sciences of the Ukrainian SSR
Splenocytes and isolated mouse spleen B lymphocytes were activated in culture by: antibodies to μ-chains of mouse immunoglobulins, Fab and F(ab')2 fragments of such antibodies, LPS from E. coli and interleukin 2(IL-2). Cell activation was monitored by [3H]-incorporation and by flow cytofluorimetry. The latter permitted detecting cell-cycle phases distribution of cells stained by acridine orange. It was shown that anti-μ antibodies were cytotoxic to mouse splenocytes and B lymphocytes. Isolated B cell could not be stimulated by Fab or F(ab')2 anti-μ, fragments but were activated by LPS and by F(ab')2 + IL-2, though F(ab')2 were stimulative to mouse splenocytes. In most cases activated cells passed G1 phase and were arrested at the GrS boundary. LPS or F(ab')2 anti-μ, addition to isolated B cells in culture caused a Ca2+-dependent increase in cGMP but not cAMP levels in these cells. Methylene bisphosphonic acid being administered to mice 24 hours before B cells isolation did not change cAMP or cGMP levels in LPS of F(ab')2 activated B lymphocytes. These data show that during the first minutes of B cell activation by LPS or F(ab')2 anti-μ, fragments there occurs a Ca2+-dependent increase in the cGMP level which is not sufficient for these cells proliferation and progression through the whole cell-cycle. The cells are usually arrested in G1 phase.
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