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. 1965 Oct 1;122(4):823–839. doi: 10.1084/jem.122.4.823

STUDIES ON RABBIT LYMPHOCYTES IN VITRO

III. PROTEIN, RNA, AND DNA SYNTHESIS BY LYMPHOCYTE CULTURES AFTER STIMULATION WITH PHYTOHAEMAGGLUTININ, WITH STAPHYLOCOCCAL FILTRATE, WITH ANTIALLOTYPE SERUM, AND WITH HETEROLOGOUS ANTISERUM TO RABBIT WHOLE SERUM

Stewart Sell 1, David S Rowe 1, P G H Gell 1
PMCID: PMC2138086  PMID: 4954762

Abstract

In vitro cultures of the peripheral blood lymphocytes of rabbits may be stimulated with phytohaemagglutinin, staphylococcal filtrate, antiallotype serum, or sheep anti-rabbit whole serum to synthesize protein, RNA and DNA as indicated by the incorporation of radiolabelled precursor substances into these products. A sequence of events found in all stimulated cultures characteristically shows protein synthesis followed by RNA synthesis, histologic blast transformation, DNA synthesis, and mitosis, with the complete sequence requiring 48 hours. All four stimulants induce essentially identical metabolic changes. Characterization of the proteins synthesized by lymphocytes in vitro has failed to demonstrate immunoglobulin synthesis by stimulated or non-stimulated cultures. It is concluded that the majority of proteins produced by peripheral lymphocytes stimulated in vitro are most likely cellular proteins related to the metabolic alterations necessary for mitosis. Absorption of sheep antisera to whole rabbit serum with rabbit IgG does not always remove the transforming capacity of the sheep antisera. Thus, it is likely that antibodies to proteins other than IgG present in the small lymphocyte may also be able to stimulate transformation. A possible common mechanism for the induction of lymphoblast transformation may be the ability of both specific and non-specific stimulants to react with protein constituents of the lymphocyte which may also be present in serum.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BACH F., HIRSCHHORN K. GAMMA-GLOBULIN PRODUCTION BY HUMAN LYMPHOCYTES IN VITRO. Exp Cell Res. 1963 Dec;32:592–595. doi: 10.1016/0014-4827(63)90197-6. [DOI] [PubMed] [Google Scholar]
  2. BECKMAN L. Effect of phytohaemagglutinin on human serum and cell proteins. Nature. 1962 Aug 11;195:582–583. doi: 10.1038/195582a0. [DOI] [PubMed] [Google Scholar]
  3. COULSON A. S., CHALMERS D. G. EFFECTS OF PHYTOHAEMAGGLUTININ ON LEUCOCYTES. Lancet. 1964 Oct 10;2(7363):819–820. doi: 10.1016/s0140-6736(64)90598-7. [DOI] [PubMed] [Google Scholar]
  4. HARRIS H., ROBSON E. B. Precipitin reactions between extracts of seeds of Canavalia ensiformis (Jack Bean) and normal and pathological serum proteins. Vox Sang. 1963 May-Jun;8:348–355. [PubMed] [Google Scholar]
  5. HAYHOE F. G., QUAGLINO D. AUTORADIOGRAPHIC INVESTIGATIONS OF RNA AND DNA METABOLISM OF HUMAN LEUCOCYTES CULTURED WITH PHYTOHAEMAGGLUTININ; URIDINE-5-3H AS A SPECIFIC PRECURSOR OF RNA. Nature. 1965 Jan 9;205:151–154. doi: 10.1038/205151a0. [DOI] [PubMed] [Google Scholar]
  6. HULLIGER L., SORKIN E. Synthesis of antibodies by blood leucocytes of the rabbit. Nature. 1963 Apr 20;198:299–299. doi: 10.1038/198299a0. [DOI] [PubMed] [Google Scholar]
  7. Kelus A. S., Gell P. G. An allotypic determinant specific to rabbit macroglobulin. Nature. 1965 Apr 17;206(981):313–314. doi: 10.1038/206313b0. [DOI] [PubMed] [Google Scholar]
  8. LANDY M., SANDERSON R. P., BERNSTEIN M. T., JACKSON A. L. ANTIBODY PRODUCTION BY LEUCOCYTES IN PERIPHERAL BLOOD. Nature. 1964 Dec 26;204:1320–1321. doi: 10.1038/2041320a0. [DOI] [PubMed] [Google Scholar]
  9. LING N. R., SPICER E., JAMES K., WILLIAMSON N. THE ACTIVATION OF HUMAN PERIPHERAL LYMPHOCYTES BY PRODUCTS OF STAPHYLOCOCCI. Br J Haematol. 1965 Jul;11:421–431. doi: 10.1111/j.1365-2141.1965.tb06604.x. [DOI] [PubMed] [Google Scholar]
  10. MACKINNEY A. A., Jr, STOHLMAN F., Jr, BRECHER G. The kinetics of cell proliferation in cultures of human peripheral blood. Blood. 1962 Mar;19:349–358. [PubMed] [Google Scholar]
  11. ROBBINS J. H. TISSUE CULTURE STUDIES OF THE HUMAN LYMPHOCYTE. Science. 1964 Dec 25;146(3652):1648–1654. doi: 10.1126/science.146.3652.1648. [DOI] [PubMed] [Google Scholar]
  12. SCHEIDEGGER J. J. Une micro-méthode de l'immuno-electrophorèse. Int Arch Allergy Appl Immunol. 1955;7(2):103–110. [PubMed] [Google Scholar]
  13. SELL S., GELL P. G. STUDIES ON RABBIT LYMPHOCYTES IN VITRO. I. STIMULATION OF BLAST TRANSFORMATION WITH AN ANTIALLOTYPE SERUM. J Exp Med. 1965 Aug 1;122:423–440. doi: 10.1084/jem.122.2.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. TAO T. W. PHYTOHEMAGGLUTININ ELICITATION OF SPECIFIC ANAMNESTIC IMMUNE RESPONSE IN VITRO. Science. 1964 Oct 9;146(3641):247–248. doi: 10.1126/science.146.3641.247. [DOI] [PubMed] [Google Scholar]

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