Biopolym. Cell. 1988; 4(6):303-309.
Cell Biology
Mouse B lymphocyte activation of proliferation and cyclic nucleotides at early stages of activation
1Komissarenko S. V.
  1. A. V. Palladin Institute of Biochemistry, Academy of Sciences of the Ukrainian SSR
    Kiev, USSR


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.


[1] Ashcroft FM. Ion channels in lymphocytes. Immunol Today. 1984; 5(8):232-233.
[2] Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19-25;308(5961):693-8.
[3] Bijsterbosch MK, Meade CJ, Turner GA, Klaus GG. B lymphocyte receptors and polyphosphoinositide degradation. Cell. 1985;41(3):999-1006.
[4] Gemsa D, Leser HG, Seitz M, Deimann W, B?rlin E. Membrane perturbation and stimulation of arachidonic acid metabolism. Mol Immunol. 1982;19(10):1287-96.
[5] Olashaw N., Pledger W. Mechanisms initiating cellular proliferation. Mediators in cell growth and differentiation. Eds R. Ford, A. Maizel. New York: Raven press, 1985:31-44.
[6] Campisi J, Gray HE, Pardee AB, Dean M, Sonenshein GE. Cell-cycle control of c-myc but not c-ras expression is lost following chemical transformation. Cell. 1984;36(2):241-7.
[7] Lukinov DA, Komissarenko SV, Tikhonova SN, Alekseeva TA, Danko IM, Poletaev AI, Gnuchev NV. The content of cyclic nucleotides in different cell cycle phases of plasmacytoma cells and influence of exogenous nucleotides on the cell distribution in the cell cycle. Doklady Akad Nauk Ukr SSR. Ser B. 1986; (2):69-71.
[8] Parker C W. Control of lymphocyte function. N Engl J Med. 1976;295(21):1180-6.
[9] Hadden JW, Coffey RG, Ananthakrishnan R, Hadden EM. Cyclic nucleotides and calcium in lymphocyte regulation and activation. Ann N Y Acad Sci. 1979;332:241-54.
[10] Nestler EJ, Greengard P. Protein phosphorylation in the brain. Nature. 1983 Oct 13-19;305(5935):583-8.
[11] Zak KP, Komissarenko SV, Butenko ZA, Gruzov MA, Karlova NP, Khomenko BM, Vinnitskaya ML. Ultrastructure of different populations immunoglobulin with lymphocytes labeled with PAP complex topography and immunoglobulins on the cell membrane surface. Molekulyarnaya biologiya. 1982; Is. 33:11-15.
[12] Coffey R. G. Assays for cyclic nucleotides including clinical applications. Immunopharmacology. Eds J. W. Hadden et al. New York: Plenum press, 1977:389-412.
[13] Komissarenko SV, Avrameas S. Properties of immunoadsorbents prepared by antigen coupling to glutaraldehydeactivated polyacrylamide gel, BrCN-activated Sepharose and by copolymerization of antigens by glutaraldehyde. Ukr Biokhim Zh. 1978;50(4):500-11.
[14] Darzynkiewicz Z. Acridine orange as a molecular probe in studies of nucleic acids in situ. Flow cytometry and sorting . Eds M. R. Melamed et al. New York: John Wiley and Sons, 1979. 285-316.
[15] Sieckmann DG, Asofsky R, Mosier DE, Zitron IM, Paul WE. Activation of mouse lymphocytes by anti-immunoglobulin. I. Parameters of the proliferative response. J Exp Med. 1978;147(3):814-29.
[16] Parker DC. Stimulation of mouse lymphocytes by insoluble anti-mouse immunoglobulin. Nature. 1975;258(5533):361-3.
[17] Melchers F, Andersson J. Factors controlling the B-cell cycle. Annu Rev Immunol. 1986;4:13-36.
[18] Komissarenko SV, Karlova NP, Kolesnikova IN et al. Application diphosphonates as immunomodulators. Immunoregulators Chemistry and Biology. Riga, Zinatne, 1985; 237-52.
[19] Komissarenko S. V., Hadden J. W. Cyclic nucleotides and cNMP dependent protein kinases in lymphocyte activation. Abstr. Vth Int. Congr. Immunol. Kvoto, 1983:25.