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Sustained localized expression of ligand for the activating NKG2D receptor impairs natural cytotoxicity in vivo and reduces tumor immunosurveillance

Abstract

Upregulation of the inducible gene products MICA (human) and Rae-1 (mouse) may promote tumor surveillance and autoimmunity by engaging the activating receptor NKG2D on natural killer (NK) cells and T cells. Nevertheless, sustained expression of MICA by tumors can also elicit NKG2D downregulation, perhaps indicating 'immunoevasion'. Investigating this paradox, we report here that constitutive Rae-1ε transgene expression in normal epithelium elicited local and systemic NKG2D downregulation, generalized but reversible defects in NK cell–mediated cytotoxicity and mild CD8+ T cell defects. The extent of NKG2D downregulation correlated well with the incidence and progression of cutaneous carcinogenesis, emphasizing the utility of NKG2D as a marker of tumor resistance. Thus, NKG2D engagement is a natural mediator of immunosurveillance, which can be compromised by locally sustained ligand expression but potentially restored by innate immune activation.

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Figure 1: Transgenic Rae-1ε protein expression from an epithelium-specific or globally active promotor.
Figure 2: NKG2D is downregulated in vivo in Rae-1-transgenic mice to amounts comparable to those of Rae-1+ tumor-bearing mice.
Figure 3: Defects in Rae-1-targeted natural cytotoxicity in vivo.
Figure 4: In vivo stimulation of NK cells by poly(I:C) restores impaired natural cytotoxicity responses to Rae-1+ targets and MHClo targets.
Figure 5: Effective generation of antigen-specific memory cytotoxic T cells in Rae-1-transgenic mice despite downregulated NKG2D.
Figure 6: Increased tumor susceptibility in Rae-1-transgenic mice.

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Acknowledgements

We thank C. Brooks (University of Newcastle), J. Dyson and E. Simpson (Medical Research Council Unit, Hammersmith Hospital), and members of the laboratory for advice and assistance; M. Allen for help with immunohistology and inducible transgenics; R. Ahmed (Emory), J. Shires and D. Barber for lymphocytic choriomeningitis virus infection; F. Watt for an involucrin promoter; G. Bates for the chicken β-actin promoter; and A. Steinle for sharing data before publication. This paper is dedicated to the memory of J. Scheckner. Supported by the Wellcome Trust (D.E.O. and A.C.H.) and the National Institutes of Health (R.E.T. and M.G.).

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Correspondence to Adrian C Hayday.

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Supplementary information

Supplementary Fig. 1

Generating Rae-1 transgenic mice. (PDF 2723 kb)

Supplementary Fig. 2

Rae-1 transgene expression in lymphocyte populations (PDF 4487 kb)

Supplementary Fig. 3

Specific loss of NKG2D staining in Rae-1 transgenic mice. (PDF 5029 kb)

Supplementary Fig. 4

Rae-1 is sufficient to drive NKG2D downregulation in vitro. (PDF 6952 kb)

Supplementary Fig. 5

In vivo natural cytotoxicity to Rae-1 targets is dependent on NK cells but not αβ T cells. (PDF 2120 kb)

Supplementary Fig. 6

HY-specific CD8 memory T cell cytotoxicity in Rae-1 transgenic mice. (PDF 1727 kb)

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Oppenheim, D., Roberts, S., Clarke, S. et al. Sustained localized expression of ligand for the activating NKG2D receptor impairs natural cytotoxicity in vivo and reduces tumor immunosurveillance. Nat Immunol 6, 928–937 (2005). https://doi.org/10.1038/ni1239

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