Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1979 Jun 1;149(6):1460–1476. doi: 10.1084/jem.149.6.1460

The in vitro generation and sustained culture of nude mouse cytolytic T- lymphocytes

PMCID: PMC2184891  PMID: 156241

Abstract

In addition to allowing for the long-term culture of both murine and human cytolytic T lymphocytes, T-cell growth factor (TCGF) functions as the key proliferation-inducing second signal in both T-cell antigen sensitization and mitogenesis. The observation that thymocytes responded normally to T-cell mitogens in the presence of TCGF, prompted the investigation of the effect of TCGF on nude mouse lymphocyte responses in vitro. We found that spleen, lymph node, and bone marrow cells, isolated from nude mice, were incapable of producing TCGF yet responded normally to T-cell mitogen sensitization provided stimulation was conducted in the presence of TCGF. Nude mouse spleen cells were also capable of responding to alloantigen sensitization in mixed lymphocyte cultures (NLMC) conducted in the presence of TCGF. Thy-1 antigen-positive cells harvested from TCGF-supplemented nude mouse MLC effectively mediated the cytolysis of alloantigen-specific target cells as tested in standard 51Cr-release assays. Cytolytic nude mouse effector cells have remained in TCGF-dependent culture for over 3 mo during which they have continued to mediate significant levels of alloantigen-specific cytolytic reactivity. These results suggest that prothymocytes present in nude mice are capable of responding to immunologic stimuli by differentiating, in vitro, into cytolytic T lymphocytes and that furthermore, a major function of the thymus may be to effect the maturation of TCGF-producing cells.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

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

  1. Armerding D., Katz D. H. Activation of T and B lymphocytes in vitro. IV. Regulatory influence on specific T cell functions by a thymus extract factor. J Immunol. 1975 Apr;114(4):1248–1254. [PubMed] [Google Scholar]
  2. Bach J. F., Dardenne M., Goldstein A. L., Guha A., White A. Appearance of T-cell markers in bone marrow rosette-forming cells after incubation with thymosin, a thymic hormone. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2734–2738. doi: 10.1073/pnas.68.11.2734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker P. E., Gillis S., Ferm M. M., Smith K. A. The effect of T cell growth factor on the generation of cytolytic T cells. J Immunol. 1978 Dec;121(6):2168–2173. [PubMed] [Google Scholar]
  4. Cerottini J. C., Nordin A. A., Brunner K. T. Specific in vitro cytotoxicity of thymus-derived lymphocytes sensitized to alloantigens. Nature. 1970 Dec 26;228(5278):1308–1309. doi: 10.1038/2281308a0. [DOI] [PubMed] [Google Scholar]
  5. Feldmann M., Wagner H., Basten A., Holmes M. Humoral and cell mediated responses in vitro of spleen cells from mice with thymic aplasia (nude mice). Aust J Exp Biol Med Sci. 1972 Oct;50(5):651–660. doi: 10.1038/icb.1972.57. [DOI] [PubMed] [Google Scholar]
  6. Gershwin M. E., Ikeda R. M., Kruse W. L., Wilson F., Shifrine M., Spangler W. Age-dependent loss in New Zealand mice of morphological and functional characteristics of thymic epithelial cells. J Immunol. 1978 Mar;120(3):971–979. [PubMed] [Google Scholar]
  7. Gillis S., Baker P. E., Ruscetti F. W., Smith K. A. Long-term culture of human antigen-specific cytotoxic T-cell lines. J Exp Med. 1978 Oct 1;148(4):1093–1098. doi: 10.1084/jem.148.4.1093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gillis S., Ferm M. M., Ou W., Smith K. A. T cell growth factor: parameters of production and a quantitative microassay for activity. J Immunol. 1978 Jun;120(6):2027–2032. [PubMed] [Google Scholar]
  9. Gillis S., Smith K. A. In vitro generation of tumor-specific cytotoxic lymphocytes. Secondary allogeneic mixed tumor lymphocyte culture of normal murine spleen cells. J Exp Med. 1977 Aug 1;146(2):468–482. doi: 10.1084/jem.146.2.468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gillis S., Smith K. A. Long term culture of tumour-specific cytotoxic T cells. Nature. 1977 Jul 14;268(5616):154–156. doi: 10.1038/268154a0. [DOI] [PubMed] [Google Scholar]
  11. Goldstein A. L., Low T. L., McAdoo M., McClure J., Thurman G. B., Rossio J., Lai C. Y., Chang D., Wang S. S., Harvey C. Thymosin alpha1: isolation and sequence analysis of an immunologically active thymic polypeptide. Proc Natl Acad Sci U S A. 1977 Feb;74(2):725–729. doi: 10.1073/pnas.74.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hensen E. J., Hoefsmit E. C., Van Den Tweel J. G. Augmentation of mitogen responsiveness in human lymphocytes by a humoral factor obtained from thymic epithelial cultures. Clin Exp Immunol. 1978 May;32(2):309–317. [PMC free article] [PubMed] [Google Scholar]
  13. Irlé C., Piguet P. F., Vassalli P. In vitro maturation of immature thymocytes into immunocompetent T cells in the absence of direct thymic influence. J Exp Med. 1978 Jul 1;148(1):32–45. doi: 10.1084/jem.148.1.32. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kindred B., Corley R. B. A T cell-replacing factor specific for histocompatibility antigens in mice. Nature. 1977 Aug 11;268(5620):531–532. doi: 10.1038/268531a0. [DOI] [PubMed] [Google Scholar]
  15. Komuro K., Boyse E. A. In-vitro demonstration of thymic hormone in the mouse by conversion of precursor cells into lymphocytes. Lancet. 1973 Apr 7;1(7806):740–743. doi: 10.1016/s0140-6736(73)92127-2. [DOI] [PubMed] [Google Scholar]
  16. Komuro K., Boyse E. A. Induction of T lymphocytes from precursor cells in vitro by a product of the thymus. J Exp Med. 1973 Aug 1;138(2):479–482. doi: 10.1084/jem.138.2.479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lonai P., Mogilner B., Rotter V., Trainin N. Studies on the effect of a thymic humoral factor on differentiation of thymus-derived lymphocytes. Eur J Immunol. 1973 Jan;3(1):21–26. doi: 10.1002/eji.1830030106. [DOI] [PubMed] [Google Scholar]
  18. Miller H. C., Schmiege S. K., Rule A. Production of functional T cells after treatment of bone marrow with thymic factor. J Immunol. 1973 Oct;111(4):1005–1009. [PubMed] [Google Scholar]
  19. Nabholz M., Engers H. D., Collavo D., North M. Cloned T-cell lines with specific cytolytic activity. Curr Top Microbiol Immunol. 1978;81:176–187. doi: 10.1007/978-3-642-67448-8_29. [DOI] [PubMed] [Google Scholar]
  20. Pantelouris E. M. Observations on the immunobiology of 'nude' mice. Immunology. 1971 Feb;20(2):247–252. [PMC free article] [PubMed] [Google Scholar]
  21. Piguet P. F., Vassalli P. Rejection of allo-or xenografts of lymphoid cells by nude mice: T cell suicide as a result of cooperation between histoincompatible T and B cells. J Immunol. 1978 Jan;120(1):79–85. [PubMed] [Google Scholar]
  22. Pritchard H., Micklem H. S. Haemopoietic stem cells and progenitors of functional T-lymphocytes in the bone marrow of 'nude' mice. Clin Exp Immunol. 1973 Aug;14(4):597–607. [PMC free article] [PubMed] [Google Scholar]
  23. Rosenberg S. A., Schwarz S., Spiess P. J. In vitro growth of murine T cells. II. Growth of in vitro sensitized cells cytotoxic for alloantigens. J Immunol. 1978 Nov;121(5):1951–1955. [PubMed] [Google Scholar]
  24. Scheid M. P., Hoffmann M. K., Komuro K., Hämmerling U., Abbott J., Boyse E. A., Cohen G. H., Hooper J. A., Schulof R. S., Goldstein A. L. Differentiation of T cells induced by preparations from thymus and by nonthymic agents. J Exp Med. 1973 Oct 1;138(4):1027–1032. doi: 10.1084/jem.138.4.1027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Small M., Trainin N. Increase in antibody-forming cells ofneonatally thymectomized mice receiving calf thymus extract. Nature. 1967 Oct 28;216(5113):377–379. doi: 10.1038/216377a0. [DOI] [PubMed] [Google Scholar]
  26. Strausser J. L., Rosenberg S. A. In vitro growth of cytotoxic human lymphocytes. I. Growth of cells sensitized in vitro to alloantigens. J Immunol. 1978 Oct;121(4):1491–1495. [PubMed] [Google Scholar]
  27. Trainin N., Small M. Studies on some physicochemical properties of a thymus humoral factor conferring immunocompetence on lymphoid cells. J Exp Med. 1970 Nov;132(5):885–897. doi: 10.1084/jem.132.5.885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wagner H. The correlation between the proliferative and the cytotoxic responses of mouse lymphocytes to allogeneic cells in vitro. J Immunol. 1972 Sep;109(3):630–637. [PubMed] [Google Scholar]
  29. Waksal S. D., Cohen I. R., Waksal H. W., Wekerle H., St Pierre R. L., Feldman M. Induction of T-cells differentiation in vitro by thymus epithelial cells. Ann N Y Acad Sci. 1975 Feb 28;249:492–498. doi: 10.1111/j.1749-6632.1975.tb29098.x. [DOI] [PubMed] [Google Scholar]
  30. Willis-Carr J. I., Ochs H. D., Wedgwood R. J. Induction of T-lymphocyte differentiation by thymic epithelial cell monolayers. Clin Immunol Immunopathol. 1978 Jul;10(3):315–324. doi: 10.1016/0090-1229(78)90187-3. [DOI] [PubMed] [Google Scholar]
  31. Wortis H. H., Nehlsen S., Owen J. J. Abnormal development of the thymus in "nude" mice. J Exp Med. 1971 Sep 1;134(3 Pt 1):681–692. doi: 10.1084/jem.134.3.681. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES