Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1972 Jun 1;135(6):1363–1374. doi: 10.1084/jem.135.6.1363

A QUANTITATIVE ASSAY FOR THE PROGENITORS OF BONE MARROW-ASSOCIATED LYMPHOCYTES

Louis Lafleur 1, R G Miller 1, R A Phillips 1
PMCID: PMC2139178  PMID: 4554453

Abstract

A cell transfer assay system was developed to study the precursors of bone marrow-associated (B) lymphocytes in the adult mouse. The rationale of the assay is to inject into irradiated mice a cell suspension depleted of B lymphocytes, to wait a period of time to let precursor cells differentiate to B lymphocytes, then to correlate the number of B cells present in the recipient mice with the number of precursor cells injected. The assay as described was shown to be linear in the range of 105–3 x 106 fractionated bone marrow cells. Kinetic studies indicated that precursor cells start producing detectable numbers of B cells within 3 days after transplantation; B cell activity then increases with a doubling time of 24 hr. Physical characterization of that precursor cell has shown that it is lighter and sediments faster than small lymphocytes. Precursor cells were found in bone marrow and spleen but could not be detected in peripheral lymph nodes. Results of physical analysis also indicate that the precursors of B lymphocytes described here may not be pluripotent stem cells for the immune system.

Full Text

The Full Text of this article is available as a PDF (651.1 KB).

Selected References

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

  1. Burleson R., Levey R. H. Demonstration of thymic function in vitro. Transplant Proc. 1971 Mar;3(1):918–922. [PubMed] [Google Scholar]
  2. Edwards G. E., Miller R. G., Phillips R. A. Differentiation of rosette-forming cells from myeloid stem cells. J Immunol. 1970 Sep;105(3):719–729. [PubMed] [Google Scholar]
  3. FORD C. E., HAMERTON J. L., BARNES D. W., LOUTIT J. F. Cytological identification of radiation-chimaeras. Nature. 1956 Mar 10;177(4506):452–454. doi: 10.1038/177452a0. [DOI] [PubMed] [Google Scholar]
  4. Gorczynski R. M., Miller R. G., Phillips R. A. Homogeneity of antibody-producing cells as analysed by their buoyant density in gradients of Ficoll. Immunology. 1970 Nov;19(5):817–829. [PMC free article] [PubMed] [Google Scholar]
  5. Gorczynski R. M., Miller R. G., Phillips R. A. Identification by density separation of antigen-specific surface receptors on the progenitors of antibody-producing cells. Immunology. 1971 May;20(5):693–705. [PMC free article] [PubMed] [Google Scholar]
  6. Gorczynski R. M., Miller R. G., Phillips R. A. In vivo requirement for a radiation-resistant cells in the immune response to sheep erythrocytes. J Exp Med. 1971 Nov 1;134(5):1201–1221. doi: 10.1084/jem.134.5.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gregory C. J., Lajtha L. G. Recovery of immune responsiveness in lethally-irradiated mice protected with syngeneic marrow cells. Int J Radiat Biol Relat Stud Phys Chem Med. 1970;17(2):117–126. doi: 10.1080/09553007014550161. [DOI] [PubMed] [Google Scholar]
  8. KENNEDY J. C., TILL J. E., SIMINOVITCH L., MCCULLOCH E. A. RADIOSENSITIVITY OF THE IMMUNE RESPONSE TO SHEEP RED CELLS IN THE MOUSE, AS MEASURED BY THE HEMOLYTIC PLAQUE METHOD. J Immunol. 1965 May;94:715–722. [PubMed] [Google Scholar]
  9. Kennedy J. C., Siminovitch L., Till J. E., McCulloch E. A. A transplantation assay for mouse cells responsive to antigenic stimulation by sheep erythrocytes. Proc Soc Exp Biol Med. 1965 Dec;120(3):868–873. doi: 10.3181/00379727-120-30678. [DOI] [PubMed] [Google Scholar]
  10. Miller J. F., Mitchell G. F. Cell to cell interaction in the immune response. V. Target cells for tolerance induction. J Exp Med. 1970 Apr 1;131(4):675–699. doi: 10.1084/jem.131.4.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Miller R. G., Phillips R. A. Sedimentation analysis of the cells in mice required to initiate an in vivo immune response to sheep erythrocytes. Proc Soc Exp Biol Med. 1970 Oct;135(1):63–67. doi: 10.3181/00379727-135-34988. [DOI] [PubMed] [Google Scholar]
  12. Miller R. G., Phillips R. A. Separation of cells by velocity sedimentation. J Cell Physiol. 1969 Jun;73(3):191–201. doi: 10.1002/jcp.1040730305. [DOI] [PubMed] [Google Scholar]
  13. Mitchell G. F., Miller J. F. Cell to cell interaction in the immune response. II. The source of hemolysin-forming cells in irradiated mice given bone marrow and thymus or thoracic duct lymphocytes. J Exp Med. 1968 Oct 1;128(4):821–837. doi: 10.1084/jem.128.4.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nossal G. J., Bussard A. E., Lewis H., Mazie J. C. In vitro stimulation of antibody formation by peritoneal cells. I. Plaque technique of high sensitivity enabling access to the cells. J Exp Med. 1970 May 1;131(5):894–916. doi: 10.1084/jem.131.5.894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nowell P. C., Hirsch B. E., Fox D. H., Wilson D. B. Evidence for the existence of multipotential lympho-hematopoietic stem cells in adult rat. J Cell Physiol. 1970 Apr;75(2):151–158. doi: 10.1002/jcp.1040750203. [DOI] [PubMed] [Google Scholar]
  16. Small M., Trainin N. Contribution of a thymic humoral factor to the development of an immunologically competent population from cells of mouse bone marrow. J Exp Med. 1971 Sep 1;134(3 Pt 1):786–800. doi: 10.1084/jem.134.3.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Talmage D. W., Radovich J., Hemmingsen H. Cell interaction in antibody synthesis. Adv Immunol. 1970;12:271–282. doi: 10.1016/s0065-2776(08)60171-6. [DOI] [PubMed] [Google Scholar]
  18. Trentin J., Wolf N., Cheng V., Fahlberg W., Weiss D., Bonhag R. Antibody production by mice repopulated with limited numbers of clones of lymphoid cell precursors. J Immunol. 1967 Jun;98(6):1326–1337. [PubMed] [Google Scholar]
  19. Unanue E. R., Grey H. M., Rabellino E., Campbell P., Schmidtke J. Immunoglobulins on the surface of lymphocytes. II. The bone marrow as the main source of lymphocytes with detectable surface-bound immunoglobulin. J Exp Med. 1971 Jun 1;133(6):1188–1198. doi: 10.1084/jem.133.6.1188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Worton R. G., McCulloch E. A., Till J. E. Physical separation of hemopoietic stem cells from cells forming colonies in culture. J Cell Physiol. 1969 Oct;74(2):171–182. doi: 10.1002/jcp.1040740209. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES