Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1970 Jan 1;131(1):57–76. doi: 10.1084/jem.131.1.57

IN VITRO AND IN VIVO STUDIES OF THE IMMUNE RESPONSE TO SHEEP ERYTHROCYTES USING PARTIALLY PURIFIED CELL PREPARATIONS

J S Haskill 1, Pauline Byrt 1, J Marbrook 1
PMCID: PMC2138767  PMID: 5409948

Abstract

The BSA density-gradient technique for separating mouse spleen cells into partially purified populations has been used to compare the responsiveness of such populations to SRBC using in vivo and in vitro techniques. Two major populations were distinguished, one of which responded very well in vivo with an exponential dose response and poorly in vitro (fraction 3), and another which responded in vivo and in vitro with a linear dose response (fraction 2). A light density, radiation-resistant component was identified which markedly stimulated the response of fraction 3 in vitro, and a density gradient profile was obtained for this cell which did not correspond with a macrophage profile. A high density, radiation-sensitive cell was identified which stimulated the response of PFC precursors in lighter regions of the gradient. The activity of this cell could be replaced using thymus cells. A density profile for the PFC precursor cell was obtained by assaying small numbers of spleen cell fractions in the presence of an excess of the two auxiliary cell types.

Full Text

The Full Text of this article is available as a PDF (896.9 KB).

Selected References

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

  1. Bussard A. E. Antibody formation in nonimmune mouse peritoneal cells after incubation in gum containing antigen. Science. 1966 Aug 19;153(3738):887–888. doi: 10.1126/science.153.3738.887. [DOI] [PubMed] [Google Scholar]
  2. Claman H. N., Chaperon E. A., Triplett R. F. Thymus-marrow cell combinations. Synergism in antibody production. Proc Soc Exp Biol Med. 1966 Aug-Sep;122(4):1167–1171. doi: 10.3181/00379727-122-31353. [DOI] [PubMed] [Google Scholar]
  3. Cunningham A. J., Szenberg A. Further improvements in the plaque technique for detecting single antibody-forming cells. Immunology. 1968 Apr;14(4):599–600. [PMC free article] [PubMed] [Google Scholar]
  4. Dutton R. W. Further studies of the stimulation of DNA synthesis in cultures of spleen cell suspensions by homologous cells in inbred strains of mice and rats. J Exp Med. 1965 Oct 1;122(4):759–770. doi: 10.1084/jem.122.4.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  6. FISHMAN M., ADLER F. L. Antibody formation initiated in vitro. II. Antibody synthesis in x-irradiated recipients of diffusion chambers containing nucleic acid derived from macrophages incubated with antigen. J Exp Med. 1963 Apr 1;117:595–602. doi: 10.1084/jem.117.4.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gallily R., Feldman M. The role of macrophages in the induction of antibody in x-irradiated animals. Immunology. 1967 Feb;12(2):197–206. [PMC free article] [PubMed] [Google Scholar]
  8. Globerson A., Auerbach R. Primary antibody response in organ cultures. J Exp Med. 1966 Nov 1;124(5):1001–1016. doi: 10.1084/jem.124.5.1001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Globerson A., Auerbach R. Primary immune reactions in organ cultures. Science. 1965 Aug 27;149(3687):991–993. doi: 10.1126/science.149.3687.991. [DOI] [PubMed] [Google Scholar]
  10. Globerson A. In vitro studies on radiation lymphoid recovery of mouse spleen. J Exp Med. 1966 Jan 1;123(1):25–32. doi: 10.1084/jem.123.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Haskill J. S. Density distribution analysis of antigen sensitive cells in the rat. Nature. 1967 Dec 23;216(5121):1229–1231. doi: 10.1038/2161229a0. [DOI] [PubMed] [Google Scholar]
  12. Haskill J. S. Density distribution analysis of antigen-sensitive cells in the rat. J Exp Med. 1969 Oct 1;130(4):877–893. doi: 10.1084/jem.130.4.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Marbrook J. Primary immune response in cultures of spleen cells. Lancet. 1967 Dec 16;2(7529):1279–1281. doi: 10.1016/s0140-6736(67)90393-5. [DOI] [PubMed] [Google Scholar]
  15. Miller J. F., Mitchell G. F. Cell to cell interaction in the immune response. I. Hemolysin-forming cells in neonatally thymectomized mice reconstituted with thymus or thoracic duct lymphocytes. J Exp Med. 1968 Oct 1;128(4):801–820. doi: 10.1084/jem.128.4.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mishell R. I., Dutton R. W. Immunization of normal mouse spleen cell suspensions in vitro. Science. 1966 Aug 26;153(3739):1004–1006. doi: 10.1126/science.153.3739.1004. [DOI] [PubMed] [Google Scholar]
  17. Mitchell G. F., Miller J. F. Immunological activity of thymus and thoracic-duct lymphocytes. Proc Natl Acad Sci U S A. 1968 Jan;59(1):296–303. doi: 10.1073/pnas.59.1.296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mosier D. E. A requirement for two cell types for antibody formation in vitro. Science. 1967 Dec 22;158(3808):1573–1575. doi: 10.1126/science.158.3808.1573. [DOI] [PubMed] [Google Scholar]
  19. Mosier D. E. Cell interactions in the primary immune response in vitro: a requirement for specific cell clusters. J Exp Med. 1969 Feb 1;129(2):351–362. doi: 10.1084/jem.129.2.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mosier D. E., Coppleson L. W. A THREE-CELL INTERACTION REQUIRED FOR THE INDUCTION OF THE PRIMARY IMMUNE RESPONSE in vitro. Proc Natl Acad Sci U S A. 1968 Oct;61(2):542–547. doi: 10.1073/pnas.61.2.542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Playfair J. H., Papermaster B. W., Cole L. J. Focal antibody production by transferred spleen cells in irradiated mice. Science. 1965 Aug 27;149(3687):998–1000. doi: 10.1126/science.149.3687.998. [DOI] [PubMed] [Google Scholar]
  22. Shortman K. The separation of different cell classes from lymphoid organs. II. The purification and analysis of lymphocyte populations by equilibrium density gradient centrifugation. Aust J Exp Biol Med Sci. 1968 Aug;46(4):375–396. doi: 10.1038/icb.1968.32. [DOI] [PubMed] [Google Scholar]
  23. Syeklocha D., Siminovitch L., Till J. E., McCulloch E. A. The proliferative state of antigen-sensitive precursors of hemolysin-producing cells, determined by the use of the inhibitor, vinblastine. J Immunol. 1966 Mar;96(3):472–477. [PubMed] [Google Scholar]
  24. Tao T. W., Uhr J. W. Primary-type antibody response in vitro. Science. 1966 Mar 4;151(3714):1096–1098. doi: 10.1126/science.151.3714.1096. [DOI] [PubMed] [Google Scholar]

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

RESOURCES