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. 1998 Jan;93(1):122–128. doi: 10.1046/j.1365-2567.1998.00394.x

Monoclonal antibody anti-NC-2 identifies a second receptor on cells mediating natural cytotoxicity in mice.

H Shirzadeh 1, R C Burton 1, J H Brien 1, Y C Smart 1
PMCID: PMC1364115  PMID: 9536128

Abstract

We have previously reported the identification by the murine monoclonal antibody (mAb) 1C4 of the first leucocyte receptor which is involved in natural cytotoxicity (NC) against WEHI-164; the NC-1.1 receptor. We report herein the identification and characterization of a second leucocyte receptor which is involved in NC, NC-2 (MW 50,000), identified by a rat anti-mouse mAb D9 (immunoglobulin G2a; IgG2a). Flow cytometric analysis showed that NC-2 was expressed on < 6% of splenic leucocytes of different inbred mouse strains and 96% of the cells of a mast-cell line which has high NC activity. In vitro treatment of splenic leucocytes with the D9 mAb blocked effector cell-WEHI-164 target cell conjugation and NC by approximately 50% without affecting natural killing (NK). Western blot analysis of affinity purified NC-2 and NC-1.1 using the D9 and 1C4 mAbs showed specific reactivity of the proteins with D9 and 1C4, respectively. Pretreatment of splenic leucocytes with both mAbs blocked NC 84%, a result which almost doubled that caused by either mAb alone. Flow cytometric screening of 16 different mouse cell lines showed that 19% of the cell lines expressed both receptors, 6% expressed only NC-2, 44% expressed mainly or only NC-1.1 and the remaining cells expressed neither receptor. These data indicate that D9 identifies a xeno-antigen, NC-2, which is expressed on cells mediating NC and not NK, and that it is not the previously described NC-1.1 allo-antigen. We conclude that NC-2 is likely to be one of a number of receptor molecules on cells mediating NC against tumour cells.

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Selected References

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

  1. Brien J. H., Smart Y. C., Farrelly M. L., Burton R. C. Phenotype and morphology of murine NC-1.1+ natural cytotoxic cells. Immunol Cell Biol. 1994 Apr;72(2):161–168. doi: 10.1038/icb.1994.24. [DOI] [PubMed] [Google Scholar]
  2. Clarke G. R., Shirzadeh H., Pang G., Beagley K. W., Burton R. C., Smart Y. C. TNF-alpha is not the sole mediator of WEHI-164 tumour cell killing in natural cytotoxicity. Cytokine. 1997 Apr;9(4):254–262. doi: 10.1006/cyto.1996.0162. [DOI] [PubMed] [Google Scholar]
  3. DUNN T. B., POTTER M. A transplantable mast-cell neoplasm in the mouse. J Natl Cancer Inst. 1957 Apr;18(4):587–601. [PubMed] [Google Scholar]
  4. Feldmann M., Boylston A., Hogg N. M. Immunological effects of IgT synthesized by theta-positive cell lines. Eur J Immunol. 1976 Jun;5(6):429–431. doi: 10.1002/eji.1830050615. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Herberman R. B., Holden H. T. Natural cell-mediated immunity. Adv Cancer Res. 1978;27:305–377. doi: 10.1016/s0065-230x(08)60936-7. [DOI] [PubMed] [Google Scholar]
  7. Holmgreen S. P., Wang X., Clarke G. R., Noltorp R. S., Roberts T. K., Burton R. C., Robinson P. J., Smart Y. C. Phosphorylation of the NC-1.1 receptor and regulation of natural cytotoxicity by protein kinase C and cyclic GMP-dependent protein kinase. J Immunol. 1997 Mar 1;158(5):2035–2041. [PubMed] [Google Scholar]
  8. Hoskin D. W., Roder J. C. Production and characterization of a novel monoclonal antibody inhibitory for murine natural killer cell activity. Immunol Invest. 1992 Feb;21(1):11–23. doi: 10.3109/08820139209069359. [DOI] [PubMed] [Google Scholar]
  9. Jutila M. A., Kroese F. G., Jutila K. L., Stall A. M., Fiering S., Herzenberg L. A., Berg E. L., Butcher E. C. Ly-6C is a monocyte/macrophage and endothelial cell differentiation antigen regulated by interferon-gamma. Eur J Immunol. 1988 Nov;18(11):1819–1826. doi: 10.1002/eji.1830181125. [DOI] [PubMed] [Google Scholar]
  10. Kiessling R., Haller O. Natural killer cells in the mouse: an alternative immune surveillance mechanism? Contemp Top Immunobiol. 1978;8:171–201. doi: 10.1007/978-1-4684-0922-2_6. [DOI] [PubMed] [Google Scholar]
  11. Kiessling R., Wigzell H. An analysis of the murine NK cell as to structure, function and biological relevance. Immunol Rev. 1979;44:165–208. doi: 10.1111/j.1600-065x.1979.tb00270.x. [DOI] [PubMed] [Google Scholar]
  12. Martiniello R., Smart Y. C., Burton R. C. Monoclonal antibody 2B6-F2 identifies a subset of murine natural killer cells with receptors for YAC-1 and WEHI-164 targets. Cell Immunol. 1994 Jun;156(1):155–169. doi: 10.1006/cimm.1994.1161. [DOI] [PubMed] [Google Scholar]
  13. Rouse B. T., Wagner H., Harris A. W. In vivo activity of in vitro immunized lymphocytes. I. Tumor allograft rejection mediated by in vitro activated mouse thymocytes. J Immunol. 1972 May;108(5):1353–1361. [PubMed] [Google Scholar]
  14. Shirzadeh H., Clarke G. R., McNeil H. P., Wang H., Burton R. C., Smart Y. C. An IL-3-induced splenic NC-1.1+ mast cell line mediates natural cytotoxicity independent of TNF-alpha. Cell Immunol. 1996 Dec 15;174(2):147–154. doi: 10.1006/cimm.1996.0304. [DOI] [PubMed] [Google Scholar]
  15. Smart Y. C., Farrelly M. L., Burton R. C. Correlation of growth of tumours in NC-cell-depleted mice with NC- and NK-cell-mediated lysis in vitro. Int J Cancer. 1992 Mar 12;50(5):817–821. doi: 10.1002/ijc.2910500526. [DOI] [PubMed] [Google Scholar]
  16. Smart Y. C., Stevenson K. L., Farrelly M. L., Brien J. H., Burton R. C. Production of a monoclonal allo-antibody to murine natural cytotoxic cells. Immunol Cell Biol. 1990 Aug;68(Pt 4):277–284. doi: 10.1038/icb.1990.38. [DOI] [PubMed] [Google Scholar]
  17. Smart Y. C., Stevenson K. L., Thorne R. F., Thomas W. D., Hsu L. H., Burton R. C. Expression of natural killer (NK) cell-specific alloantigens on a mouse NK-like cell line. Immunol Cell Biol. 1989 Aug;67(Pt 4):239–242. doi: 10.1038/icb.1989.36. [DOI] [PubMed] [Google Scholar]
  18. Stutman O., Paige C. J., Figarella E. F. Natural cytotoxic cells against solid tumors in mice. I. Strain and age distribution and target cell susceptibility. J Immunol. 1978 Nov;121(5):1819–1826. [PubMed] [Google Scholar]
  19. Unkeless J. C. Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors. J Exp Med. 1979 Sep 19;150(3):580–596. doi: 10.1084/jem.150.3.580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Yokoyama W. M. Recognition structures on natural killer cells. Curr Opin Immunol. 1993 Feb;5(1):67–73. doi: 10.1016/0952-7915(93)90083-5. [DOI] [PubMed] [Google Scholar]

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