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. 1971 Nov;21(5):767–779.

Blastogenic response of lymphocytes separated from bone marrow to allogeneic lymphoid cells in vitro

Y Yoshida, D G Osmond
PMCID: PMC1408166  PMID: 5115610

Abstract

Fractions of cells were separated from the bone marrow and spleen of Lewis rats by brief centrifugation in linear sucrose-serum density gradients and were cultured in vitro either alone or mixed with F1 (Lewis × Brown Norway) hybrid rat lymphoid cells. After 4 days the incorporation of [3H]thymidine into DNA was measured by scintillation counting, and the morphology and incidence of [3H]thymidine-labelled cells were determined in radioautographs.

Lymphocyte-rich, slowly-sedimenting fractions of Lewis rat bone marrow cells showed increased incorporation of [3H]thymidine and the development of many large proliferating blast-like cells when cultured with F1 hybrid cells. This blastogenic response was greater than could be ascribed to contaminating intravascular blood lymphocytes. Slowly-sedimenting fractions of spleen cells, consisting mainly of small lymphocytes, showed a greater blastogenic responsiveness to F1 hybrid cells than that of whole spleen, while rapidly-sedimenting spleen cell fractions, containing many large cells, showed a reduced responsiveness. Paradoxically, rapidly-sedimenting marrow cell fractions containing few small lymphocytes, showed an increment in [3H]thymidine incorporation when cultured with F1 cells but this was due to active macrophage proliferation as well as to blastogenesis.

The results demonstrate that some parenchymal bone marrow lymphocytes undergo blastogenic transformation in response to allogeneic lymphocytes in vitro.

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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. LYMPHOCYTE INTERACTION: A POTENTIAL HISTOCOMPATIBILITY TEST IN VITRO. Science. 1964 Feb 21;143(3608):813–814. doi: 10.1126/science.143.3608.813. [DOI] [PubMed] [Google Scholar]
  2. BAIN B., VAS M. R., LOWENSTEIN L. THE DEVELOPMENT OF LARGE IMMATURE MONONUCLEAR CELLS IN MIXED LEUKOCYTE CULTURES. Blood. 1964 Jan;23:108–116. [PubMed] [Google Scholar]
  3. Brahim F., Osmond D. G. Migration of bone marrow lymphocytes demonstrated by selective bone marrow labeling with thymidine-H3. Anat Rec. 1970 Oct;168(2):139–159. doi: 10.1002/ar.1091680202. [DOI] [PubMed] [Google Scholar]
  4. Cantor H., Mandel M. A., Asofsky R. Studies of thoracic duct lymphocytes of mice. II. A quantitative comparison of the capacity of thoracic duct lymphocytes and other lymphoid cells to induce graft-versus-host reactions. J Immunol. 1970 Feb;104(2):409–413. [PubMed] [Google Scholar]
  5. Davis W. E., Jr, Cole L. J., Schaffer W. T. Studies on synergistic thymus-bone marrow cell interactions in immunological responses. Proc Soc Exp Biol Med. 1970 Jan;133(1):144–150. doi: 10.3181/00379727-133-34427. [DOI] [PubMed] [Google Scholar]
  6. Dutton R. W. Symposium on in vitro studies of the immune response. II. Significance of the reaction of lymphoid cells to homologous tissue. Bacteriol Rev. 1966 Jun;30(2):397–407. doi: 10.1128/br.30.2.397-407.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. EVERETT N. B., CAFFREY R. W., RIEKE W. O. RECIRCULATION OF LYMPHOCYTES. Ann N Y Acad Sci. 1964 Feb 28;113:887–897. doi: 10.1111/j.1749-6632.1964.tb40710.x. [DOI] [PubMed] [Google Scholar]
  8. Ford W. L., Burr W., Simonsen M. A lymph node weight assay for the graft-versus-host activity of rat lymphoid cells. Transplantation. 1970 Sep;10(3):258–266. doi: 10.1097/00007890-197009000-00007. [DOI] [PubMed] [Google Scholar]
  9. Gordon J., MacLean L. D. A lymphocyte-stimulating factor produced in vitro. Nature. 1965 Nov 20;208(5012):795–796. doi: 10.1038/208795a0. [DOI] [PubMed] [Google Scholar]
  10. Haas R. J., Bohne F., Fliedner T. M. On the development of slowly-turning-over cell types in neonatal rat bone marrow (studies utilizing the complete tritiated thymidine labelling method complemented by C-14 thymidine administration). Blood. 1969 Dec;34(6):791–805. [PubMed] [Google Scholar]
  11. Kasakura S., Lowenstein L. A factor stimulating DNA synthesis derived from the medium of leukocyte cultures. Nature. 1965 Nov 20;208(5012):794–795. doi: 10.1038/208794a0. [DOI] [PubMed] [Google Scholar]
  12. Levine S. Local and regional forms of graft-versus-host disease in lymph nodes. Transplantation. 1968 Sep;6(6):799–802. doi: 10.1097/00007890-196809000-00009. [DOI] [PubMed] [Google Scholar]
  13. Linna T. J., Lidén S. Cell migration from the bone marrow to the spleen in young guinea pigs. Int Arch Allergy Appl Immunol. 1969;35(1):35–46. doi: 10.1159/000230157. [DOI] [PubMed] [Google Scholar]
  14. McGregor D. D. Bone marrow origin of immunologically competent lymphocytes in the rat. J Exp Med. 1968 May 1;127(5):953–966. doi: 10.1084/jem.127.5.953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. OSMOND D. G., EVERETT N. B. BONE MARROW BLOOD VOLUME AND TOTAL RED CELL MASS OF THE GUINEA-PIG AS DETERMINED BY 59-FE-ERYTHROCYTE DILUTION AND LIQUID NITROGEN FREEZING. Q J Exp Physiol Cogn Med Sci. 1965 Jan;50:1–14. doi: 10.1113/expphysiol.1965.sp001758. [DOI] [PubMed] [Google Scholar]
  16. OSMOND D. G., EVERETT N. B. RADIOAUTOGRAPHIC STUDIES OF BONE MARROW LYMPHOCYTES IN VIVO AND IN DIFFUSION CHAMBER CULTURES. Blood. 1964 Jan;23:1–17. [PubMed] [Google Scholar]
  17. Oppenheim J. J., Leventhal B. G., Hersh E. M. The transformation of column-purified lymphocytes with nonspecific and specific antigenic stimuli. J Immunol. 1968 Aug;101(2):262–267. [PubMed] [Google Scholar]
  18. Osmond D. G. The non-thymic origin of lymphocytes. Anat Rec. 1969 Sep;165(1):109–112. doi: 10.1002/ar.1091650114. [DOI] [PubMed] [Google Scholar]
  19. SIMONSEN M. Graft versus host reactions. Their natural history, and applicability as tools of research. Prog Allergy. 1962;6:349–467. [PubMed] [Google Scholar]
  20. Schwarz M. R. The mixed lymphocyte reaction: an in vitro test for tolerance. J Exp Med. 1968 May 1;127(5):879–890. doi: 10.1084/jem.127.5.879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schwarz M. R. Transformation of rat small lymphocytes with allogeneic lymphoid cells. Am J Anat. 1967 Nov;121(3):559–570. doi: 10.1002/aja.1001210308. [DOI] [PubMed] [Google Scholar]
  22. Silvers W. K., Wilson D. B., Palm J. Mixed leukocyte reactions and histocompatibility in rats. Science. 1967 Feb 10;155(3763):703–704. doi: 10.1126/science.155.3763.703. [DOI] [PubMed] [Google Scholar]
  23. Singhal S. K., Richter M., Osmond D. G. Cells involved in the immune response. 3. Responsiveness in antigens of a lymphocyte-rich fraction of normal rabbit bone marrow. Int Arch Allergy Appl Immunol. 1968;34(3):224–232. [PubMed] [Google Scholar]
  24. VOLKMAN A., GOWANS J. L. THE ORIGIN OF MACROPHAGES FROM BONE MARROW IN THE RAT. Br J Exp Pathol. 1965 Feb;46:62–70. [PMC free article] [PubMed] [Google Scholar]
  25. Wilson D. B., Blyth JL NOWELL P. C. Quantitative studies on the mixed lymphocyte interaction in rats. 3. Kinetics of the response. J Exp Med. 1968 Nov 1;128(5):1157–1181. doi: 10.1084/jem.128.5.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wilson D. B. Quantitative studies on the mixed lymphocyte interaction in rats. I. Conditions and parameters of response. J Exp Med. 1967 Oct 1;126(4):625–654. doi: 10.1084/jem.126.4.625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yoffey J. M., Hudson G., Osmond D. G. The lymphocyte in guinea-pig bone marrow. J Anat. 1965 Oct;99(Pt 4):841–860. [PMC free article] [PubMed] [Google Scholar]
  28. Yoshida Y., Osmond D. G. Identity and proliferation of small lymphocyte precursors in cultures of lymphocyte-rich fractions of guinea pig bone marrow. Blood. 1971 Jan;37(1):73–86. [PubMed] [Google Scholar]

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