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. 1982 Jun;36(3):1028–1035. doi: 10.1128/iai.36.3.1028-1035.1982

Production of mouse lymphotoxin by phytohemagglutinin-stimulated spleen cells requires two cell fractions.

R R Aksamit, E J Leonard
PMCID: PMC551435  PMID: 6980190

Abstract

The appearance of lymphotoxin in the culture fluid of phytohemagglutinin (PHA)-stimulated mouse spleen cells required two cell fractions that were separated by adherence to plastic. Upon stimulation with PHA, neither cell fraction alone produced significant amounts of lymphotoxin; however, when the cell fractions were combined and then stimulated with PHA, full activity was produced. Cytotoxic activity was not fully restored by combining PHA-stimulated cultured fluids from adherent and nonadherent cell fractions. This indicated that the cytotoxic activity was not the result of two factors, one produced by each cell fraction, that acted on the target cells, but rather, two cells interacted to produce lymphotoxin. Treatment of the unfractionated spleen cells with monoclonal anti-Thy1.2 and complement before PHA stimulation greatly reduced the production of lymphotoxin and indicated that at least one of the cells was a T cell. Lymphotoxin production was partially restored by the addition of nonadherent cells to the anti-Thy1.2-treated cells, suggesting that the T cell was nonadherent. Treatment of unfractionated cells with either silica or carrageenan had no effect on the subsequent production of lymphotoxin by PHA, suggesting that the adherent cell was not actively phagocytic.

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Selected References

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

  1. Adams D. O., Kao K. J., Farb R., Pizzo S. V. Effector mechanisms of cytolytically activated macrophages. II. Secretion of a cytolytic factor by activated macrophages and its relationship to secreted neutral proteases. J Immunol. 1980 Jan;124(1):293–300. [PubMed] [Google Scholar]
  2. Aksamit R. R., Kim K. J. Macrophage cell lines produce a cytotoxin. J Immunol. 1979 May;122(5):1785–1790. [PubMed] [Google Scholar]
  3. Aksamit R. R., Leonard E. J. Effect of mouse lymphotoxin on radiation-damaged cells. Cell Immunol. 1980 Jun;52(1):163–175. doi: 10.1016/0008-8749(80)90409-8. [DOI] [PubMed] [Google Scholar]
  4. Amino N., Linn E. S., Pysher T. J., Mier R., Moore G. E., DeGroot L. J. Human lymphotoxin obtained from established lymphoid lines: purification characteristics and inhibition by anti-immunoglobulin. J Immunol. 1974 Oct;113(4):1334–1345. [PubMed] [Google Scholar]
  5. Bloom B. R., Stoner G., Gaffney J., Shevach E., Green I. Production of migration inhibitory factor and lymphotoxin by non-T cells. Eur J Immunol. 1975 Mar;5(3):218–220. doi: 10.1002/eji.1830050314. [DOI] [PubMed] [Google Scholar]
  6. Boraschi D., Meltzer M. S. Defective tumoricidal capacity of macrophages from A/J mice. II. Comparison of the macrophage cytotoxic defect of A/J mice with that of lipid A-unresponsive C3H/HeJ mice. J Immunol. 1979 Apr;122(4):1592–1597. [PubMed] [Google Scholar]
  7. Durkin H. G., Bash J. A., Waksman B. H. Separation of T cell subpopulations capable of DNA synthesis, lymphotoxin release, and regulation of antigen and phytohemagglutinin responses on the basis of density and adherence properties. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5090–5094. doi: 10.1073/pnas.72.12.5090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eardley D. D., Shen F. W., Gershon R. K., Ruddle N. H. Lymphotoxin production by subsets of T cells. J Immunol. 1980 Mar;124(3):1199–1202. [PubMed] [Google Scholar]
  9. Evans C. H., Rabin E. S., DiPaolo J. A. The susceptibility of guinea pig cells to the colony-inhibitory activity of lymphotoxin during carcinogenesis. Cancer Res. 1977 Mar;37(3):898–903. [PubMed] [Google Scholar]
  10. Gately M. K., Gately C. L., Henney C. S., Mayer M. M. Studies on lymphokines: the production of antibody to guinea pig lymphotoxin and its use to distinguish lymphotoxin from migration inhibitory factor and mitogenic factor. J Immunol. 1975 Sep;115(3):817–826. [PubMed] [Google Scholar]
  11. Granger G. A., Kolb W. P. Lymphocyte in vitro cytotoxicity: mechanisms of immune and non-immune small lymphocyte mediated target L cell destruction. J Immunol. 1968 Jul;101(1):111–120. [PubMed] [Google Scholar]
  12. Hiserodt J. C., Fair D. S., Granger G. A. Identification of multiple cytolytic components associated with the beta-LT class of lymphotoxins released by mitogen-activity human lymphocytes in vitro. J Immunol. 1976 Nov;117(5 Pt 1):1503–1506. [PubMed] [Google Scholar]
  13. Hiserodt J. C., Prieur A. M., Granger G. A. In vitro lymphocyte cytotoxicity. I. Evidence of multiple cytotoxic molecules secreted by mitogen activated human lymphoid cells in vitro. Cell Immunol. 1976 Jun 15;24(2):277–288. doi: 10.1016/0008-8749(76)90212-4. [DOI] [PubMed] [Google Scholar]
  14. Hiserodt J. C., Tiangco G. J., Granger G. A. The LT system in experimental animals. I. Rapid release of high levels of lymphotoxin (LT) activity from murine lymphocytes during the interaction with lectin-treated allogeneic or xenogeneic target cells in vitro. J Immunol. 1979 Jul;123(1):311–316. [PubMed] [Google Scholar]
  15. Hiserodt J. C., Tiangco G. J., Granger G. A. The LT system in experimental animals. II. Physical and immunologic characteristics of molecules with LT activity rapidly released by murine lymphoid cells activated on lectin-coated allogeneic monolayers in vitro. J Immunol. 1979 Jul;123(1):317–324. [PubMed] [Google Scholar]
  16. Hiserodt J. C., Tiangco G. J., Granger G. A. The LT system in experimental animals. IV. Rapid specific lysis of 51CR-labeled allogeneic target cells by highly unstable high m.w. lymphotoxin-receptor complex(es) released in vitro by activated alloimmune murine T lymphocytes. J Immunol. 1979 Jul;123(1):332–341. [PubMed] [Google Scholar]
  17. Hiserodt J. C., Yamamoto R. S., Granger G. A. The human LT system. III. Characterization of a high molecular weight LT class (complex) composed of the various smaller MW LT classes and subclasses in association with Ig-like molecules. Cell Immunol. 1978 Jul;38(2):417–433. doi: 10.1016/0008-8749(78)90071-0. [DOI] [PubMed] [Google Scholar]
  18. Hiserodt J. C., Yamamoto R. S., Granger G. A. The human LT system. IV. Studies on the large MW LT complex class: association of these molecules with specific antigen binding receptor(s) in vitro. Cell Immunol. 1978 Dec;41(2):380–396. doi: 10.1016/0008-8749(78)90235-6. [DOI] [PubMed] [Google Scholar]
  19. Karnovsky M. L., Lazdins J. K. Biochemical criteria for activated macrophages. J Immunol. 1978 Sep;121(3):809–813. [PubMed] [Google Scholar]
  20. Klimpel G. R., Dean J. H., Day K. D., Chen P. B., Lucas D. O. Lymphotoxin and interferon production by rossette-separated human peripheral blood leukocytes. Cell Immunol. 1977 Aug;32(2):293–301. doi: 10.1016/0008-8749(77)90206-4. [DOI] [PubMed] [Google Scholar]
  21. Kramer J. J., Granger G. A. The in vitro induction and release of a cell toxin by immune C57B1-6 mouse peritoneal macrophages. Cell Immunol. 1972 Jan;3(1):88–100. doi: 10.1016/0008-8749(72)90229-8. [DOI] [PubMed] [Google Scholar]
  22. Lee S. C., Lucas Z. J. Regulatory factors produced by lymphocytes. I. The occurrence of multiple alpha-lymphotoxins associated with ribonuclease activity. J Immunol. 1976 Jul;117(1):283–291. [PubMed] [Google Scholar]
  23. Macfarlan R. I., White D. O. A soluble cytotoxic factor from macrophages. Aust J Exp Biol Med Sci. 1980 Oct;58(5):489–500. doi: 10.1038/icb.1980.50. [DOI] [PubMed] [Google Scholar]
  24. McMyne P. S., Strejan G. H. Relationships between cell-mediated immunity and the IgE antibody response. I. Lymphotoxin production to DNP-Ascaris conjugates. Cell Immunol. 1980 Aug 15;54(1):140–154. doi: 10.1016/0008-8749(80)90197-5. [DOI] [PubMed] [Google Scholar]
  25. Meltzer M. S., Bartlett G. L. Cytotoxicity in vitro by products of specifically stimulated spleen cells: susceptibility of tumor cells and normal cells. J Natl Cancer Inst. 1972 Nov;49(5):1439–1443. [PubMed] [Google Scholar]
  26. Männel D. N., Moore R. N., Mergenhagen S. E. Macrophages as a source of tumoricidal activity (tumor-necrotizing factor). Infect Immun. 1980 Nov;30(2):523–530. doi: 10.1128/iai.30.2.523-530.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Reed W. P., Lucas Z. J. Cytotoxic activity of lymphocytes. V. Role of soluble toxin in macrophage-inhibited cultures of tumor cells. J Immunol. 1975 Aug;115(2):395–404. [PubMed] [Google Scholar]
  28. Rosenberg S. A., Henrichon M., Coyne J. A., David J. R. Guinea pig lymphotoxin (LT). I. In vitro studies of LT produced in response to antigen stimulation of lymphocytes. J Immunol. 1973 Jun;110(6):1623–1629. [PubMed] [Google Scholar]
  29. Ross M. W., Tiangco G. J., Horn P., Hiserodt J. C., Granger G. A. The LT system in experimental animals. III. Physicochemical characteristics and relationships of lymphotoxin (LT) molecules released in vitro by activated lymphoid cells from several animal species. J Immunol. 1979 Jul;123(1):325–331. [PubMed] [Google Scholar]
  30. Ruddle N. H., Waksman B. H. Cytotoxicity mediated by soluble antigen and lymphocytes in delayed hypersensitivity. 3. Analysis of mechanism. J Exp Med. 1968 Dec 1;128(6):1267–1279. doi: 10.1084/jem.128.6.1267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rundell J. O., Evans C. H. Species specificity of guinea pig and human lymphotoxin colony inhibitory activity. Immunopharmacology. 1981 Feb;3(1):9–18. doi: 10.1016/0162-3109(81)90034-5. [DOI] [PubMed] [Google Scholar]
  32. Shacks S. J., Chiller J., Granger G. A. Studies on in vitro models of cellular immunity: the role of T and B cells in the secretion of lymphotoxin. Cell Immunol. 1973 May;7(2):313–321. doi: 10.1016/0008-8749(73)90253-0. [DOI] [PubMed] [Google Scholar]
  33. Trivers G., Braungart D., Leonard E. J. Mouse lymphotoxin. J Immunol. 1976 Jul;117(1):130–135. [PubMed] [Google Scholar]
  34. Walker S. M., Lee S. C., Lucas Z. J. Cytotoxic activity of lymphocytes. VI. Heterogeneity of cytotoxins in supernatants of mitogen-activated lymphocytes. J Immunol. 1976 Mar;116(3):807–815. [PubMed] [Google Scholar]
  35. Woody J. N., Ahmed A., Knudsen R. C., Strong D. M., Sell K. W. Human T-cell heterogeneity as delineated with a specific human thymus lymphocyte antiserum. In vitro effects on mitogen response mixed leukocyte culture, cell-mediated lymphocytotoxicity, and lymphokine production. J Clin Invest. 1975 May;55(5):956–966. doi: 10.1172/JCI108025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yano K., Lucas Z. J. Cytotoxic activity of lymphocytes. VII. cellular origin of alpha-lymphotoxin. J Immunol. 1978 Feb;120(2):385–394. [PubMed] [Google Scholar]

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