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. 1981 Aug 1;154(2):362–373. doi: 10.1084/jem.154.2.362

Quantitation and clonal isolation of cytolytic T lymphocyte precursors selectively infiltrating murine sarcoma virus-induced tumors

PMCID: PMC2186418  PMID: 6973602

Abstract

A limiting dilution mixed leukocyte-tumor cell microculture system was used to quantitate cytolytic T lymphocytes and their precursors (CTL- P), which infiltrate tumors induced by injection of Moloney sarcoma- leukemia virus (MSV-MoLV) complex into C57BL/6 mice. Leukocyte populations obtained from tumors collected on day 10 after virus injection were found to contain significantly higher frequencies of operationally defined (tumor-specific) CTL-P than either peripheral blood leukocytes (PBL) or spleen cells from the same animals. When these frequencies were normalized according to the content of Lyt-2+ T cells in each tissue, average CTL-P frequencies were found to be 1/9 in tumor-infiltrating cells vs. 1/41 in PBL. These results directly demonstrate selective accumulation of CTL-P in the tumor mass. A number of clonal isolates obtained from tumor-infiltrating leukocyte populations were expanded and studied for cytolytic activity and specificity. Of 11 isolates, 10 were found to have high cytolytic activity, leading to 50% lysis of the syngeneic MoLV-derived tumor target cells in 3.5 h at lymphocyte:target cell ratios ranging from 0.5:1 to 3.2:1. Furthermore, five randomly selected clones showed H-2 restriction by their selective lytic activity against MoLV-derived syngeneic MBL-2 target cells and their lack of activity against either MoLV-derived allogeneic (LSTRA) tumor cells or against syngeneic (EL4) or allogeneic (P815) target cells unrelated to MoLV.

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Selected References

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

  1. Brunner K. T., MacDonald H. R., Cerottini J. C. Antigenic specificity of the cytolytic T lymphocyte (CTL) response to murine sarcoma virus-induced tumors. II. Analysis of the clonal progeny of CTL precursors stimulated in vitro with syngeneic tumor cells. J Immunol. 1980 Apr;124(4):1627–1634. [PubMed] [Google Scholar]
  2. Cerottini J. C., Brunner K. T. Localization of mouse isoantigens on the cell surface as revealed by immunofluorescence. Immunology. 1967 Oct;13(4):395–403. [PMC free article] [PubMed] [Google Scholar]
  3. Cerottini J. C., Engers H. D., Macdonald H. R., Brunner T. Generation of cytotoxic T lymphocytes in vitro. I. Response of normal and immune mouse spleen cells in mixed leukocyte cultures. J Exp Med. 1974 Sep 1;140(3):703–717. doi: 10.1084/jem.140.3.703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cerottini J. C., MacDonald H. R. Limiting dilution analysis of alloantigen-reactive T lymphocytes. V. Lyt phenotype of cytolytic T lymphocyte precursors reactive against normal and mutant H-2 antigens. J Immunol. 1981 Feb;126(2):490–496. [PubMed] [Google Scholar]
  5. Chapdelaine J. M., Plata F., Lilly F. Tumors induced by murine sarcoma virus contain precursor cells capable of generating tumor-specific cytolytic T lymphocytes. J Exp Med. 1979 Jun 1;149(6):1531–1536. doi: 10.1084/jem.149.6.1531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Engers H. D., MacDonald H. R. Generation of cytolytic T lymphocytes in vitro. Contemp Top Immunobiol. 1976;5:145–190. [PubMed] [Google Scholar]
  7. Gillespie G. Y., Hansen C. B., Hoskins R. G., Russell S. W. Inflammatory cells in solid murine neoplasms. IV. Cytolytic T lymphocytes isolated from regressing or progressing Moloney sarcomas. J Immunol. 1977 Aug;119(2):564–570. [PubMed] [Google Scholar]
  8. Holden H. T., Haskill J. S., Kirchner H., Herberman R. B. Two functionally distinct anti-tumor effector cells isolated from primary murine sarcoma virus-induced tumors. J Immunol. 1976 Aug;117(2):440–446. [PubMed] [Google Scholar]
  9. Jondal M., Svedmyr E., Klein E., Singh S. Killer T cells in a Burkitt's lymphoma biopsy. Nature. 1975 May 29;255(5507):405–407. doi: 10.1038/255405a0. [DOI] [PubMed] [Google Scholar]
  10. Klein E., Becker S., Svedmyr E., Jondal M., Vánky F. Tumor infiltrating lymphocytes. Ann N Y Acad Sci. 1976;276:207–216. doi: 10.1111/j.1749-6632.1976.tb41647.x. [DOI] [PubMed] [Google Scholar]
  11. Levy J. P., Leclerc J. C. The murine sarcoma virus-induced tumor: exception or general model in tumor immunology? Adv Cancer Res. 1977;24:1–66. doi: 10.1016/s0065-230x(08)61012-x. [DOI] [PubMed] [Google Scholar]
  12. Maryanski J. L., MacDonald H. R., Cerottini J. C. Limiting dilution analysis of alloantigen-reactive T lymphocytes. IV. High frequency of cytolytic T lymphocyte precursor cells in MLC blasts separated by velocity sedimentation. J Immunol. 1980 Jan;124(1):42–47. [PubMed] [Google Scholar]
  13. Nakayama E., Shiku H., Stockert E., Oettgen H. F., Old L. J. Cytotoxic T cells: Lyt phenotype and blocking of killing activity by Lyt antisera. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1977–1981. doi: 10.1073/pnas.76.4.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Plata F., Cerottini J. C., Brunner K. T. Primary and secondary in vitro generation of cytolytic T lymphocytes in the murine sarcoma virus system. Eur J Immunol. 1975 Apr;5(4):227–233. doi: 10.1002/eji.1830050402. [DOI] [PubMed] [Google Scholar]
  15. Plata F., MacDonald H. R., Sordat B. Studies on the distribution and origin of cytolytic T lymphocytes present in mice bearing Moloney murine sarcoma virus (MSV)-induced tumors. Bibl Haematol. 1975 Oct;(43):274–277. doi: 10.1159/000399147. [DOI] [PubMed] [Google Scholar]
  16. Plata F., Sordat B. Murine sarcoma virus (MSV)-induced tumors in mice. I. Distribution of MSV-immune cytolytic T lymphocytes in vivo. Int J Cancer. 1977 Feb 15;19(2):205–211. doi: 10.1002/ijc.2910190210. [DOI] [PubMed] [Google Scholar]
  17. Ryser J. E., Cerottini J. C., Brunner K. T. Generation of cytolytic T lymphocytes in vitro. IX. induction of secondary CTL responses in primary long-term MLC by supernatants from secondary MLC. J Immunol. 1978 Feb;120(2):370–377. [PubMed] [Google Scholar]
  18. Smith K. A. T-cell growth factor. Immunol Rev. 1980;51:337–357. doi: 10.1111/j.1600-065x.1980.tb00327.x. [DOI] [PubMed] [Google Scholar]
  19. Taswell C. Limiting dilution assays for the determination of immunocompetent cell frequencies. I. Data analysis. J Immunol. 1981 Apr;126(4):1614–1619. [PubMed] [Google Scholar]
  20. Weiss A., Brunner K. T., MacDonald H. R., Cerottini J. C. Antigenic specificity of the cytolytic T lymphocyte response to murine sarcoma virus-induced tumors. III. Characterization of cytolytic T lymphocyte clones specific for Moloney leukemia virus-associated cell surface antigens. J Exp Med. 1980 Nov 1;152(5):1210–1225. doi: 10.1084/jem.152.5.1210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Weiss A., MacDonald H. R., Cerottini J. C., Brunner K. T. Inhibition of cytolytic T lymphocyte clones reactive with Moloney leukemia virus-associated antigens by monoclonal antibodies: a direct approach to the study of H-2 restriction. J Immunol. 1981 Feb;126(2):482–485. [PubMed] [Google Scholar]
  22. Yron I., Wood T. A., Jr, Spiess P. J., Rosenberg S. A. In vitro growth of murine T cells. V. The isolation and growth of lymphoid cells infiltrating syngeneic solid tumors. J Immunol. 1980 Jul;125(1):238–245. [PubMed] [Google Scholar]

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