Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Ligands for the murine NKG2D receptor: expression by tumor cells and activation of NK cells and macrophages

Nature Immunology volume 1, pages 119–126 (2000)Cite this article

Abstract

Natural killer (NK) cells attack tumor and infected cells, but the receptors and ligands that stimulate them are poorly understood. Here we report the expression cloning of two murine ligands for the lectin-like receptor NKG2D. The two ligands, H-60 and Rae1β, are distant relatives of major histocompatibility complex class I molecules. NKG2D ligands are not expressed by most normal cells but are up-regulated on numerous tumor cells. We show that mouse NKG2D is expressed by NK cells, activated CD8+ T cells and activated macrophages. Expression of either NKG2D ligand by target cells triggers NK cell cytotoxicity and interferon-γ secretion by NK cells, as well as nitric oxide release and tumor necrosis factor α transcription by macrophages. Thus, through their interaction with NKG2D, H-60 and Rae1β are newly identified potent stimulators of innate immunity.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Numerous tumor cells express ligands for mNKG2D.
Figure 2: BALB/c thymocytes and stimulated splenocytes express ligands for mNKG2D.
Figure 3: The MHC I–related molecules Rae1β and H-60 are ligands for mNKG2D.
Figure 4: The NKG2D receptor is expressed on freshly isolated and IL-2–activated NK cells, CD8+ T cells and LPS-stimulated macrophages.
Figure 5: NKG2D mRNA is expressed in naïve NK cells, activated CD8+ T cells and LPS-stimulated macrophages.
Figure 6: Expression of Rae1 or H-60 induces target cell killing and IFN-γ production by NK cells.
Figure 7: Expression of Rae1 or H-60 induces nitric oxide production and TNF-α transcription in stimulated peritoneal macrophages.

Similar content being viewed by others

References

  1. Trinchieri, G. Biology of natural killer cells. Adv. Immunol. 47, 187–376 (1989).

    Article  CAS  Google Scholar 

  2. Ljunggren, H.G. & Karre, K. In search of the ‘missing self’: MHC molecules and NK cell recognition. Immunol. Today 11, 237–244 (1990).

    Article  CAS  Google Scholar 

  3. Lanier, L.L. NK cell receptors. Annu. Rev. Immunol. 16, 359–393 (1998).

    Article  CAS  Google Scholar 

  4. Vitale, M. et al. NKp44, a novel triggering surface molecule specifically expressed by activated natural killer cells, is involved in non-major histocompatibility complex-restricted tumor cell lysis. J. Exp. Med. 187 , 2065–2072 (1998).

    Article  CAS  Google Scholar 

  5. Sivori, S. et al. NKp46 is the major triggering receptor involved in the natural cytotoxicity of fresh or cultured human NK cells. Correlation between surface density of NKp46 and natural cytotoxicity against autologous, allogeneic or xenogeneic target cells. Eur. J. Immunol. 29, 1656–66 (1999).

    Article  CAS  Google Scholar 

  6. Pende, D. et al. Identification and molecular characterization of NKp30, a novel triggering receptor involved in natural cytotoxicity mediated by human natural killer cells. J. Exp. Med. 190, 1505– 16 (1999).

    Article  CAS  Google Scholar 

  7. Miyazaki, T., Dierich, A., Benoist, C. & Mathis, D. Independent modes of natural killing distinguished in mice lacking Lag3. Science 272, 405–8 ( 1996).

    Article  CAS  Google Scholar 

  8. Ryan, J., Niemi, E., Nakamura, M. & Seaman, W. NKR-P1A is a target-specific receptor that activates natural killer cell cytotoxicity. J. Exp. Med. 181, 1911–1915 ( 1995).

    Article  CAS  Google Scholar 

  9. Brown, M.H. et al. 2B4, the natural killer and T cell immunoglobulin superfamily surface protein, is a ligand for CD48. J. Exp. Med. 188, 2083–90 (1998).

    Article  CAS  Google Scholar 

  10. Houchins, J.P., Yabe, T., McSherry, C., Miyokawa, N. & Bach, F.H. Isolation and characterization of NK cell or NK/T cell-specific cDNA clones. J. Mol. Cell. Immunol. 4 , 295–304, 305–6 (1990).

    CAS  PubMed  Google Scholar 

  11. Houchins, J.P., Yabe, T., McSherry, C. & Bach, F.H. DNA sequence analysis of NKG2, a family of related cDNA clones encoding type II integral membrane proteins on human natural killer cells. J. Exp. Med. 173, 1017–20 (1991).

    Article  CAS  Google Scholar 

  12. Vance, R.E., Jamieson, A.M. & Raulet, D.H. Recognition of the class Ib molecule Qa-1(b) by putative activating receptors CD94/NKG2C and CD94/NKG2E on mouse natural killer cells . J. Exp. Med. 190, 1801– 12 (1999).

    Article  CAS  Google Scholar 

  13. Lazetic, S., Chang, C., Houchins, J.P., Lanier, L.L. & Phillips, J.H. Human natural killer cell receptors involved in MHC class I recognition are disulfide-linked heterodimers of CD94 and NKG2 subunits. J. Immunol. 157, 4741 –4745 (1996).

    CAS  PubMed  Google Scholar 

  14. Wu, J. et al. An activating immunoreceptor complex formed by NKG2D and DAP10 . Science 285, 730–2 (1999).

    Article  CAS  Google Scholar 

  15. Bauer, S. et al. Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science 285, 727– 9 (1999).

    Article  CAS  Google Scholar 

  16. Groh, V. et al. Cell stress-regulated human major histocompatibility complex class I gene expressed in gastrointestinal epithelium. Proc. Natl Acad. Sci. USA 93, 12445–50 (1996).

    Article  CAS  Google Scholar 

  17. Groh, V. et al. Broad tumor-associated expression and recognition by tumor-derived γδ T cells of MICA and MICB. Proc. Natl Acad. Sci. USA 96, 6879–84 (1999).

    Article  CAS  Google Scholar 

  18. Crowley, M.P. et al. A population of murine γδ T cells that recognize an inducible MHC class Ib molecule. Science 287, 314–6 (2000).

    Article  CAS  Google Scholar 

  19. Crowley, M.P., Reich, Z., Mavaddat, N., Altman, J.D. & Chien, Y. The recognition of the nonclassical major histocompatibility complex (MHC) class I molecule, T 10, by the γδ T cell, G8. J. Exp. Med. 185, 1223–30 (1997).

    Article  CAS  Google Scholar 

  20. Bahram, S., Bresnahan, M., Geraghty, D.E. & Spies, T. A second lineage of mammalian major histocompatibility complex class I genes . Proc. Natl Acad. Sci. USA 91, 6259– 63 (1994).

    Article  CAS  Google Scholar 

  21. Ho, A.S. et al. A receptor for interleukin 10 is related to interferon receptors . Proc. Natl Acad. Sci. USA 90, 11267– 71 (1993).

    Article  CAS  Google Scholar 

  22. Malarkannan, S. et al. The molecular and functional characterization of a dominant minor H antigen, H60. J. Immunol. 161, 3501 –9 (1998).

    CAS  PubMed  Google Scholar 

  23. Nomura, M., Takihara, Y. & Shimada, K. Isolation and characterization of retinoic acid-inducible cDNA clones in F9 cells: one of the early inducible clones encodes a novel protein sharing several highly homologous regions with a Drosophila polyhomeotic protein. Differentiation 57, 39– 50 (1994).

    Article  CAS  Google Scholar 

  24. Nomura, M. et al. Genomic structures and characterization of Rae1 family members encoding GPI-anchored cell surface proteins and expressed predominantly in embryonic mouse brain. J. Biochem. 120, 987–95 (1996).

    Article  CAS  Google Scholar 

  25. Zou, Z., Nomura, M., Takihara, Y., Yasunaga, T. & Shimada, K. Isolation and characterization of retinoic acid-inducible cDNA clones in F9 cells: a novel cDNA family encodes cell surface proteins sharing partial homology with MHC class I molecules. J. Biochem. 119, 319–28 ( 1996).

    Article  CAS  Google Scholar 

  26. Vance, R.E., Tanamachi, D.M., Hanke, T. & Raulet, D.H. Cloning of a mouse homolog of CD94 extends the family of C-type lectins on murine natural killer cells. Eur. J. Immunol. 27, 3236–3241 (1997).

    Article  CAS  Google Scholar 

  27. Ho, E.L. et al. Murine Nkg2d and Cd94 are clustered within the natural killer complex and are expressed independently in natural killer cells. Proc. Natl Acad. Sci. USA 95, 6320– 6325 (1998).

    Article  CAS  Google Scholar 

  28. Hong, W.K. & Sporn, M.B. Recent advances in chemoprevention of cancer. Science 278, 1073– 7 (1997).

    Article  CAS  Google Scholar 

  29. Amadou, C. et al. The mouse major histocompatibility complex: some assembly required. Immunol. Rev. 167, 211– 21 (1999).

    Article  CAS  Google Scholar 

  30. Cosman, D. et al. The human cytomegalovirus (HCMV) glycoprotein, UL16, binds to the MHC class I-related protein, MICB/PERB11, and to two novel, MHC class I-related molecules, ULBP1 and ULBP2. FASEB J. 14, A1018 (2000).

  31. Chalupny, J. et al. Soluble forms of the novel MHC class I–related molecules, ULBP1 and ULBP2, bind to, and functionally activate NK cells. FASEB J. 14, A1018 (2000).

  32. Vance, R.E., Kraft, J.R., Altman, J.D., Jensen, P.E. & Raulet, D.H. Mouse CD94/NKG2A is a natural killer cell receptor for the nonclassical MHC class I molecule Qa-1b. J. Exp. Med. 188, 1841–1848 (1998).

    Article  CAS  Google Scholar 

  33. Altman, J.D. et al. Phenotypic analysis of antigen-specific T lymphocytes. Science 274, 94–6 ( 1996).

    Article  CAS  Google Scholar 

  34. Elliott, J.F. et al. Genes for Plasmodium falciparum surface antigens cloned by expression in COS cells. Proc. Natl Acad. Sci. USA 87, 6363–7 (1990).

    Article  CAS  Google Scholar 

  35. Hirt, B. Selective extraction of polyoma DNA from infected mouse cell cultures. J. Molec. Biol. 26, 365–369 (1967).

    Article  CAS  Google Scholar 

  36. Kang, J. et al. T cell receptor γ gene regulatory sequences prevent the function of a novel TCRγ/p Tα pre-T cell receptor. Immunity 8, 713–721 ( 1998).

    Article  CAS  Google Scholar 

  37. Diefenbach, A. et al. Type 1 interferon (IFNαβ) and type 2 nitric oxide synthase regulate the innate immune response to a protozoan parasite. Immunity 8, 77–87 ( 1998).

    Article  CAS  Google Scholar 

  38. Förster, E. An improved general method to generate internal standards for competitive PCR. Biotechniques 16, 18– 20 (1994).

    PubMed  Google Scholar 

  39. Hanke, T. et al. Direct assessment of MHC class I binding by seven Ly49 inhibitory NK cell receptors. Immunity 11, 67– 77 (1999).

    Article  CAS  Google Scholar 

  40. Ranganath, S. et al. GATA-3–dependent enhancer activity in IL-4 gene regulation . J. Immunol. 161, 3822– 6 (1998).

    CAS  PubMed  Google Scholar 

  41. Pear, W.S., Nolan, G.P., Scott, M.L. & Baltimore, D. Production of high-titer helper-free retroviruses by transient transfection. Proc. Natl Acad. Sci. USA 90, 8392– 6 (1993).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank R.E.Vance and C.W. McMahon for advice and comments on the manuscript, J. Egen for help with the immunofluorescence staining, H. Nolla for cell sorting, C.White for expert technical assistance, J.F. Elliott, M. Fasso, L. Mendoza for their gifts of reagents and cell lines. Supported by grants from the National Institute of Health (D.H.R.) and the Deutsche Forschungsgemeinschaft (A.D.). A.D. is a Physician Postdoctoral fellow of the Howard Hughes Medical Institute.

Author information

Authors and Affiliations

  1. Department of Molecular and Cell Biology and Cancer Research Laboratory, University of California, Berkeley, USA

    Andreas Diefenbach, Amanda M. Jamieson, Scot D. Liu, Nilabh Shastri & David H. Raulet

Authors
  1. Andreas Diefenbach
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amanda M. Jamieson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Scot D. Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nilabh Shastri
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. David H. Raulet
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to David H. Raulet.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Diefenbach, A., Jamieson, A., Liu, S. et al. Ligands for the murine NKG2D receptor: expression by tumor cells and activation of NK cells and macrophages. Nat Immunol 1, 119–126 (2000). https://doi.org/10.1038/77793

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/77793

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing