Skip to main content
NCBI home page
Immunology logoLink to Immunology
. 1980 Aug;40(4):571–577.

The in vitro discrimination of rat immune and natural cytotoxicity by serum choice.

V J Lindsay, R A Allardyce
PMCID: PMC1458097  PMID: 7429543

Abstract

Natural in vitro cytotoxicity of non-immune lymphoid cells against a variety of transformed and normal iso- and allogeneic target cells has been described in a number of species (for review see Herberman, Djeu, Kay, Ortaldo, Riccardi, Bonnard, Holden, Fagnani, Santoni & Puccetti, 1979). This background cytotoxicity is a hindrance to the estimation of immune lymphocyte cytolysis (Herberman & Oldham, 1975). Other workers have demonstrated that the choice of serum in which effector (EC) or target cells (TC) are prepared or tested may greatly influence the degree of killing (Irie, Irie & Morton, 1974; Zielske & Golub, 1976; Akira & Takasugi, 1977; Ortaldo, Bonnard & Herberman, 1977). It was found that high levels of rat normal effector cell (NEC) cytotoxicity against normal rat fibroblasts observed in the presence of foetal calf serum (FCS) were markedly reduced by the inclusion of normal rat serum (NRS). This effect on NEC activity was consistently noted in eleven batches of FCS and eight batches of NRS, whereas, normal human serum (NHuS) gave variable results. Decreased TC lysis was proportional to the concentration of NRS, was independent of heat inactivation and was primarily due to an effect on the EC population. The effects of FCS and NRS were not affected by prior absorption on TC or EC nor was NEC activity related to a proliferative response of EC during the 18-48 h microcytotoxicity assay employed in these studies. Since the inclusion of NRS did not markedly diminish the levels of immune effector cell cytotoxicity we have been able consistently to demonstrate that the use of 10% NRS in long-term rat cytotoxicity assays provides a means of dissociating the measurement of immune and normal lymphoid cell cytotoxicity.

Full text

PDF
571

Selected References

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

  1. Akira D., Takasugi M. Loss of specific natural cell-mediated cytotoxicity with absorption of natural antibodies from serum. Int J Cancer. 1977 Jun 15;19(6):747–755. doi: 10.1002/ijc.2910190603. [DOI] [PubMed] [Google Scholar]
  2. Herberman R. B., Djeu J., Kay H. D., Ortaldo J. R., Riccardi C., Bonnard G. D., Holden H. T., Fagnani R., Santoni A., Puccetti P. Natural killer cells: characteristics and regulation of activity. Immunol Rev. 1979;44:43–70. doi: 10.1111/j.1600-065x.1979.tb00267.x. [DOI] [PubMed] [Google Scholar]
  3. Herberman R. B., Oldham R. K. Problems associated with study of cell-mediated immunity to human tumors by microcytotoxicity assays. J Natl Cancer Inst. 1975 Oct;55(4):749–753. doi: 10.1093/jnci/55.4.749. [DOI] [PubMed] [Google Scholar]
  4. Heslop B. F. Histocompatibility antigens in the rat: the AS2 strain in relation to the AS, BS and HS strains. Aust J Exp Biol Med Sci. 1968 Aug;46(4):479–491. doi: 10.1038/icb.1968.41. [DOI] [PubMed] [Google Scholar]
  5. Irie R. F., Irie K., Morton D. L. Characteristics of heterologous membrane antigen on cultured human cells. J Natl Cancer Inst. 1974 Dec;53(6):1545–1551. [PubMed] [Google Scholar]
  6. Koide Y., Takasugi M. Determination of specificity in natural cell-mediated cytotoxicity by natural antibodies. J Natl Cancer Inst. 1977 Oct;59(4):1099–1105. doi: 10.1093/jnci/59.4.1099. [DOI] [PubMed] [Google Scholar]
  7. Le Mevel B. P., Oldham R. K., Wells S. A., Herberman R. B. An evaluation of 125I-iododeoxyuridine as a cellular label for in vitro assays: kinetics of incorporation and toxicity. J Natl Cancer Inst. 1973 Nov;51(5):1551–1558. doi: 10.1093/jnci/51.5.1551. [DOI] [PubMed] [Google Scholar]
  8. Nunn M. E., Djeu J. Y., Glaser M., Lavrin D. H., Herberman R. B. Natural cytotoxic reactivity of rat lymphocytes against syngeneic Gross virus-induced lymphoma. J Natl Cancer Inst. 1976 Feb;56(2):393–399. doi: 10.1093/jnci/56.2.393. [DOI] [PubMed] [Google Scholar]
  9. Nunn M. E., Herberman R. B. Natural cytotoxicity of mouse, rat, and human lymphocytes against heterologous target cells. J Natl Cancer Inst. 1979 Apr;62(4):765–771. [PubMed] [Google Scholar]
  10. Oehler J. R., Herberman R. B. Natural cell-mediated cytotoxicity in rats. III. Effects of immunopharmacologic treatments on natural reactivity and on reactivity augmented by polyinosinic-polycytidylic acid. Int J Cancer. 1978 Feb 15;21(2):221–229. doi: 10.1002/ijc.2910210214. [DOI] [PubMed] [Google Scholar]
  11. Oehler J. R., Lindsay L. R., Nunn M. E., Herberman R. B. Natural cell-mediated cytotoxicity in rats. I. Tissue and strain distribution, and demonstration of a membrance receptor for the Fc portion of IgG. Int J Cancer. 1978 Feb 15;21(2):204–209. doi: 10.1002/ijc.2910210212. [DOI] [PubMed] [Google Scholar]
  12. Oehler J. R., Lindsay L. R., Nunn M. E., Holden H. T., Herberman R. B. Natural cell-mediated cytotoxicity in rats. II. In vivo augmentation of NK-cell activity. Int J Cancer. 1978 Feb 15;21(2):210–220. doi: 10.1002/ijc.2910210213. [DOI] [PubMed] [Google Scholar]
  13. Ortaldo J. R., Bonnard G. D., Herberman R. B. Cytotoxic reactivity of human lymphocytes cultured in vitro. J Immunol. 1977 Oct;119(4):1351–1357. [PubMed] [Google Scholar]
  14. Ortaldo J. R., Bonnard G. D., Kind P. D., Herberman R. B. Cytotoxicity by cultured human lymphocytes: characteristics of effector cells and specificity of cytotoxicity. J Immunol. 1979 Apr;122(4):1489–1494. [PubMed] [Google Scholar]
  15. Parker G. A., Hyatt C., Rosenberg S. A. Normal adult murine cells in tissue culture express fetal antigens. Transplantation. 1977 Feb;23(2):161–163. doi: 10.1097/00007890-197702000-00010. [DOI] [PubMed] [Google Scholar]
  16. Pierce C. W., Kapp J. A. Regulation of immune responses by suppressor T cells. Tohoku J Exp Med. 1976;5:91–143. [PubMed] [Google Scholar]
  17. Saksela E., Timonen T., Ranki A., Häyry P. Morphological and functional characterization of isolated effector cells responsible for human natural killer activity to fetal fibroblasts and to cultured cell line targets. Immunol Rev. 1979;44:71–123. doi: 10.1111/j.1600-065x.1979.tb00268.x. [DOI] [PubMed] [Google Scholar]
  18. Seeger R. C., Owen J. J. Measurement of tumor immunity in vitro with 125 I-iododeoxyuridine-labeled target cells. Transplantation. 1973 Apr;15(4):404–408. doi: 10.1097/00007890-197304000-00010. [DOI] [PubMed] [Google Scholar]
  19. Stark O., Kren V., Günther E. RtH-1 antigens in 39 rat strains and six congenic lines. Transplant Proc. 1971 Mar;3(1):165–168. [PubMed] [Google Scholar]
  20. Takasugi M., Klein E. A microassay for cell-mediated immunity. Transplantation. 1970 Mar;9(3):219–227. doi: 10.1097/00007890-197003000-00005. [DOI] [PubMed] [Google Scholar]
  21. Zielske J. V., Golub S. H. Fetal calf serum-induced blastogenic and cytotoxic responses of human lymphocytes. Cancer Res. 1976 Oct;36(10):3842–3846. [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES