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. 1968 Nov 30;128(6):1237–1254. doi: 10.1084/jem.128.6.1237

CYTOTOXICITY MEDIATED BY SOLUBLE ANTIGEN AND LYMPHOCYTES IN DELAYED HYPERSENSITIVITY

I. CHARACTERIZATION OF THE PHENOMENON

Nancy H Ruddle 1, Byron H Waksman 1
PMCID: PMC2138572  PMID: 4176937

Abstract

In the presence of specific antigen, lymph node cells from inbred rats with delayed hypersensitivity to tuberculoprotein, bovine gammaglobulin, and egg albumin produced progressive destruction of monolayers of rat embryo fibroblasts in tissue culture, first apparent at 48 hr and maximal at 72 hr. The effect was specific and did not depend on a genetic difference between the lymph node cells and target cells. It required antigen concentrations equal to or greater than 1.25 µg/ml and lymphocyte: target cell ratios of approximately 10 or 20:1. It could be evaluated both by a plaquing technique and by cell enumeration with an electronic particle counter.

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Selected References

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

  1. AL ASKARI S., DAVID J. R., LAWRENCE H. S., THOMAS L. IN VITRO STUDIES ON HOMOGRAFT SENSITIVITY. Nature. 1965 Feb 27;205:916–917. doi: 10.1038/205916a0. [DOI] [PubMed] [Google Scholar]
  2. Atkins E., Bodel P., Francis L. Release of an endogenous pyrogen in vitro from rabbit mononuclear cells. J Exp Med. 1967 Aug 1;126(2):357–384. doi: 10.1084/jem.126.2.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett B., Bloom B. R. Reactions in vivo and in vitro produced by a soluble substance associated with delayed-type hypersensitivity. Proc Natl Acad Sci U S A. 1968 Mar;59(3):756–762. doi: 10.1073/pnas.59.3.756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bennett B., Bloom B. R. Studies on the migration inhibitory factor associated with delayed-type hypersensitivity: cytodynamics and specificity. Transplantation. 1967 Jul;5(4 Suppl):996–1000. [PubMed] [Google Scholar]
  5. Bloom B. R., Bennett B. Mechanism of a reaction in vitro associated with delayed-type hypersensitivity. Science. 1966 Jul 1;153(3731):80–82. doi: 10.1126/science.153.3731.80. [DOI] [PubMed] [Google Scholar]
  6. Brunner K. T., Mauel J., Schindler R. In vitro studies of cell-bound immunity; cloning assay of the cytotoxic action of sensitized lymphoid cells on allogeneic target cells. Immunology. 1966 Nov;11(5):499–506. [PMC free article] [PubMed] [Google Scholar]
  7. CARPENTER R. R., BRANDRISS M. W. IN VITRO STUDIES OF CELLULAR HYPERSENSITIVITY. II. RELATIONSHIP OF DELAYED HYPERSENSITIVITY AND INHIBITION OF CELL MIGRATION BY PICRYLATED PROTEINS. J Exp Med. 1964 Dec 1;120:1231–1243. doi: 10.1084/jem.120.6.1231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. CARPENTER R. R. IN VITRO STUDIES OF CELLULAR HYPERSENSITIVITY. I. SPECIFIC INHIBITION OF MIGRATION OF CELLS FROM ADJUVANT-IMMUNIZED ANIMALS BY PURIFIED PROTEIN DERIVATIVE AND OTHER PROTEIN ANTIGENS. J Immunol. 1963 Dec;91:803–818. [PubMed] [Google Scholar]
  9. Caron G. A. Duration of exposure of lymphocytes to antigen in vitro needed to induce blast transformation. Int Arch Allergy Appl Immunol. 1967;32(1):98–103. doi: 10.1159/000229919. [DOI] [PubMed] [Google Scholar]
  10. DAVID J. R., AL-ASKARI S., LAWRENCE H. S., THOMAS L. DELAYED HYPERSENSITIVITY IN VITRO. I. THE SPECIFICITY OF INHIBITION OF CELL MIGRATION BY ANTIGENS. J Immunol. 1964 Aug;93:264–273. [PubMed] [Google Scholar]
  11. DAVID J. R., LAWRENCE H. S., THOMAL L. DELAYED HYPERSENSITIVITY IN VITRO. II. EFFECT OF SENSITIVE CELLS ON NORMAL CELLS IN THE PRESENCE OF ANTIGEN. J Immunol. 1964 Aug;93:274–278. [PubMed] [Google Scholar]
  12. DAVID J. R., LAWRENCE H. S., THOMAS L. DELAYED HYPERSENSITIVITY IN VITRO. 3. THE SPECIFICITY OF HAPTEN-PROTEIN CONJUGATES IN THE INHIBITION OF CELL MIGRATION. J Immunol. 1964 Aug;93:279–282. [PubMed] [Google Scholar]
  13. GANGAROSA E. J., INGLEFIELD J. T., THOMAS C. G., MORGAN H. R. Studies on hypersensitivity of human tissues in vitro. I. Tuberculin hypersensitivity. J Exp Med. 1955 Oct 1;102(4):425–433. doi: 10.1084/jem.102.4.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. GEORGE M., VAUGHAN J. H. In vitro cell migration as a model for delayed hypersensitivity. Proc Soc Exp Biol Med. 1962 Nov;111:514–521. doi: 10.3181/00379727-111-27841. [DOI] [PubMed] [Google Scholar]
  15. Glasgow L. A. Leukocytes and interferon in the host response to viral infections. II. Enhanced interferon response of leukocytes from immune animals. J Bacteriol. 1966 Jun;91(6):2185–2191. doi: 10.1128/jb.91.6.2185-2191.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Holm G., Perlmann P. Cytotoxic potential of stimulated human lymphocytes. J Exp Med. 1967 Apr 1;125(4):721–736. doi: 10.1084/jem.125.4.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. KAPRAL F. A., STEINEBRING W. R. The effect of glucose on the tuberculin reaction in tissue culture. Am Rev Tuberc. 1958 Nov;78(5):712–724. doi: 10.1164/artpd.1958.78.5.712. [DOI] [PubMed] [Google Scholar]
  18. Kasakura S., Lowenstein L. DNA and RNA synthesis and the formation of blastogenic factor in mixed leucocyte cultures. Nature. 1967 Jul 1;215(5096):80–81. doi: 10.1038/215080a0. [DOI] [PubMed] [Google Scholar]
  19. Kniazeff A. J., Rimer V., Gaeta L. Gamma-globulin foetal bovine sera: significance in virology. Nature. 1967 May 20;214(5090):805–806. doi: 10.1038/214805b0. [DOI] [PubMed] [Google Scholar]
  20. MADIN S. H., DARBY N. B., Jr Established kidney cell lines of normal adult bovine and ovine origin. Proc Soc Exp Biol Med. 1958 Jul;98(3):574–576. doi: 10.3181/00379727-98-24111. [DOI] [PubMed] [Google Scholar]
  21. Mackaness G. B., Blanden R. V. Cellular immunity. Prog Allergy. 1967;11:89–140. [PubMed] [Google Scholar]
  22. Mills J. A. The immunologic significance of antigen induced lymphocyte transformation in vitro. J Immunol. 1966 Aug;97(2):239–247. [PubMed] [Google Scholar]
  23. Moen J. K., Swift H. F. TISSUE CULTURE STUDIES ON BACTERIAL HYPERSENSITIVITY : I. TUBERCULIN SENSITIVE TISSUES. J Exp Med. 1936 Aug 31;64(3):339–353. doi: 10.1084/jem.64.3.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Möller G., Möller E. Plaque-formation by non-immune and x-irradiated lymphoid cells on monolayers of mouse embryo cells. Nature. 1965 Oct 16;208(5007):260–263. doi: 10.1038/208260a0. [DOI] [PubMed] [Google Scholar]
  25. PALM J. SEROLOGICAL DETECTION OF HISTOCOMPATIBILITY ANTIGENS IN TWO STRAINS OF RATS. Transplantation. 1964 Sep;2:603–612. doi: 10.1097/00007890-196409000-00001. [DOI] [PubMed] [Google Scholar]
  26. Pincus W. B. Cell-free cytotoxic fluids from tuberculin-treated guinea pigs. J Reticuloendothel Soc. 1967 May;4(2):140–150. [PubMed] [Google Scholar]
  27. ROSENAU W., MOON H. D. Lysis of homologous cells by sensitized lymphocytes in tissue culture. J Natl Cancer Inst. 1961 Aug;27:471–483. [PubMed] [Google Scholar]
  28. Ruddle N. H., Waksman B. H. Cytotoxic effect of lymphocyte-antigen interaction in delayed hypersensitivity. Science. 1967 Sep 1;157(3792):1060–1062. doi: 10.1126/science.157.3792.1060. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Ruddle N. H., Waksman B. H. Cytotoxicity mediated by soluble antigen and lymphocytes in delayed hypersensitivity. II. Correlation of the in vitro response with skin reactivity. J Exp Med. 1968 Dec 1;128(6):1255–1265. doi: 10.1084/jem.128.6.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. SHEA J. D., MORGAN H. R. Delayed hypersensitivity in vitro. I. Effects of tuberculoproteins on tissue from sensitive guinea pigs. Proc Soc Exp Biol Med. 1957 Mar;94(3):436–438. doi: 10.3181/00379727-94-22969. [DOI] [PubMed] [Google Scholar]
  32. STEFFEN C., ROSAK M. IN VITRO DEMONSTRATION OF ANTI-OVALBUMIN SPECIFICITY OF LYMPH NODE CELLS IN DELAYED TYPE HYPERSENSITIVITY. J Immunol. 1963 Mar;90:337–346. [PubMed] [Google Scholar]
  33. WAKSMAN B. H., MATOLTSY M. The effect of tuberculin on peritoneal exudate cells of sensitized guinea pigs in surviving cell culture. J Immunol. 1958 Sep;81(3):220–234. [PubMed] [Google Scholar]
  34. WINKLER G. F. IN VITRO DEMYELINATION OF PERIPHERAL NERVE INDUCED WITH SENSITIZED CELLS. Ann N Y Acad Sci. 1965 Mar 31;122:287–296. doi: 10.1111/j.1749-6632.1965.tb20213.x. [DOI] [PubMed] [Google Scholar]
  35. Wilson D. B. Lymphocytes as mediators of cellular immunity: destruction of homologous target cells in culture. Transplantation. 1967 Jul;5(4 Suppl):986–988. [PubMed] [Google Scholar]
  36. Wilson D. B. QUANTITATIVE STUDIES ON THE BEHAVIOR OF SENSITIZED LYMPHOCYTES IN VITRO : I. RELATIONSHIP OF THE DEGREE OF DESTRUCTION OF HOMOLOGOUS TARGET CELLS TO THE NUMBER OF LYMPHOCYTES AND TO THE TIME OF CONTACT IN CULTURE AND CONSIDERATION OF THE EFFECTS OF ISOIMMUNE SERUM. J Exp Med. 1965 Jul 1;122(1):143–166. doi: 10.1084/jem.122.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]

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