Article | Published:

Blockade of the checkpoint receptor TIGIT prevents NK cell exhaustion and elicits potent anti-tumor immunity


Checkpoint blockade enhances effector T cell function and has elicited long-term remission in a subset of patients with a broad spectrum of cancers. TIGIT is a checkpoint receptor thought to be involved in mediating T cell exhaustion in tumors; however, the relevance of TIGIT to the dysfunction of natural killer (NK) cells remains poorly understood. Here we found that TIGIT, but not the other checkpoint molecules CTLA-4 and PD-1, was associated with NK cell exhaustion in tumor-bearing mice and patients with colon cancer. Blockade of TIGIT prevented NK cell exhaustion and promoted NK cell–dependent tumor immunity in several tumor-bearing mouse models. Furthermore, blockade of TIGIT resulted in potent tumor-specific T cell immunity in an NK cell–dependent manner, enhanced therapy with antibody to the PD-1 ligand PD-L1 and sustained memory immunity in tumor re-challenge models. This work demonstrates that TIGIT constitutes a previously unappreciated checkpoint in NK cells and that targeting TIGIT alone or in combination with other checkpoint receptors is a promising anti-cancer therapeutic strategy.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Additional information

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


  1. 1.

    Lanier, L. L. NK cell recognition. Annu. Rev. Immunol. 23, 225–274 (2005).

  2. 2.

    Zhang, Q. F. et al. Liver-infiltrating CD11bCD27 NK subsets account for NK-cell dysfunction in patients with hepatocellular carcinoma and are associated with tumor progression. Cell. Mol. Immunol. 14, 819–829 (2017).

  3. 3.

    Long, E. O., Kim, H. S., Liu, D., Peterson, M. E. & Rajagopalan, S. Controlling natural killer cell responses: integration of signals for activation and inhibition. Annu. Rev. Immunol. 31, 227–258 (2013).

  4. 4.

    Chen, L. & Flies, D. B. Molecular mechanisms of T cell co-stimulation and co-inhibition. Nat. Rev. Immunol. 13, 227–242 (2013).

  5. 5.

    Pardoll, D. M. The blockade of immune checkpoints in cancer immunotherapy. Nat. Rev. Cancer 12, 252–264 (2012).

  6. 6.

    Zou, W., Wolchok, J. D. & Chen, L. PD-L1 (B7-H1) and PD-1 pathway blockade for cancer therapy: mechanisms, response biomarkers, and combinations. Sci. Transl. Med. 8, 328rv4 (2016).

  7. 7.

    Sharma, P. & Allison, J. P. Immune checkpoint targeting in cancer therapy: toward combination strategies with curative potential. Cell 161, 205–214 (2015).

  8. 8.

    Yu, X. et al. The surface protein TIGIT suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. Nat. Immunol. 10, 48–57 (2009).

  9. 9.

    Stanietsky, N. et al. The interaction of TIGIT with PVR and PVRL2 inhibits human NK cell cytotoxicity. Proc. Natl. Acad. Sci. USA 106, 17858–17863 (2009).

  10. 10.

    Bi, J. et al. T-cell Ig and ITIM domain regulates natural killer cell activation in murine acute viral hepatitis. Hepatology 59, 1715–1725 (2014).

  11. 11.

    Bi, J. et al. TIGIT safeguards liver regeneration through regulating natural killer cell-hepatocyte crosstalk. Hepatology 60, 1389–1398 (2014).

  12. 12.

    Joller, N. et al. Treg cells expressing the coinhibitory molecule TIGIT selectively inhibit proinflammatory Th1 and Th17 cell responses. Immunity 40, 569–581 (2014).

  13. 13.

    Johnston, R. J. et al. The immunoreceptor TIGIT regulates antitumor and antiviral CD8+ T cell effector function. Cancer Cell 26, 923–937 (2014).

  14. 14.

    Chauvin, J. M. et al. TIGIT and PD-1 impair tumor antigen-specific CD8+ T cells in melanoma patients. J. Clin. Invest. 125, 2046–2058 (2015).

  15. 15.

    Kong, Y. et al. T-cell immunoglobulin and ITIM domain (TIGIT) associates with CD8+ T cell exhaustion and poor clinical outcome in AML patients. Clin. Cancer Res. 22, 3057–3066 (2016).

  16. 16.

    Shibuya, A. et al. DNAM-1, a novel adhesion molecule involved in the cytolytic function of T lymphocytes. Immunity 4, 573–581 (1996).

  17. 17.

    Chan, C. J. et al. The receptors CD96 and CD226 oppose each other in the regulation of natural killer cell functions. Nat. Immunol. 15, 431–438 (2014).

  18. 18.

    Kurtulus, S. et al. TIGIT predominantly regulates the immune response via regulatory T cells. J. Clin. Invest. 125, 4053–4062 (2015).

  19. 19.

    van Elsas, A., Hurwitz, A. A. & Allison, J. P. Combination immunotherapy of B16 melanoma using anti-cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and granulocyte/macrophage colony-stimulating factor (GM-CSF)-producing vaccines induces rejection of subcutaneous and metastatic tumors accompanied by autoimmune depigmentation. J. Exp. Med. 190, 355–366 (1999).

  20. 20.

    Yang, Y. F. et al. Enhanced induction of antitumor T-cell responses by cytotoxic T lymphocyte-associated molecule-4 blockade: the effect is manifested only at the restricted tumor-bearing stages. Cancer Res. 57, 4036–4041 (1997).

  21. 21.

    Quezada, S. A., Peggs, K. S., Curran, M. A. & Allison, J. P. CTLA4 blockade and GM-CSF combination immunotherapy alters the intratumor balance of effector and regulatory T cells. J. Clin. Invest. 116, 1935–1945 (2006).

  22. 22.

    Li, B. et al. Anti-programmed death-1 synergizes with granulocyte macrophage colony-stimulating factor–secreting tumor cell immunotherapy providing therapeutic benefit to mice with established tumors. Clin. Cancer Res. 15, 1623–1634 (2009).

  23. 23.

    Curran, M. A. & Allison, J. P. Tumor vaccines expressing flt3 ligand synergize with ctla-4 blockade to reject preimplanted tumors. Cancer Res. 69, 7747–7755 (2009).

  24. 24.

    Curran, M. A., Montalvo, W., Yagita, H. & Allison, J. P. PD-1 and CTLA-4 combination blockade expands infiltrating T cells and reduces regulatory T and myeloid cells within B16 melanoma tumors. Proc. Natl. Acad. Sci. USA 107, 4275–4280 (2010).

  25. 25.

    Ngiow, S. F. et al. Anti-TIM3 antibody promotes T cell IFN-γ-mediated antitumor immunity and suppresses established tumors. Cancer Res. 71, 3540–3551 (2011).

  26. 26.

    Dong, J., McPherson, C. M. & Stambrook, P. J. Flt-3 ligand: a potent dendritic cell stimulator and novel antitumor agent. Cancer Biol. Ther. 1, 486–489 (2002).

  27. 27.

    Hou, S. et al. Eradication of hepatoma and colon cancer in mice with Flt3L gene therapy in combination with 5-FU. Cancer Immunol. Immunother. 56, 1605–1613 (2007).

  28. 28.

    Salmon, H. et al. Expansion and activation of CD103+ dendritic cell progenitors at the tumor site enhances tumor responses to therapeutic PD-L1 and BRAF inhibition. Immunity 44, 924–938 (2016).

  29. 29.

    Zou, W. Immunosuppressive networks in the tumour environment and their therapeutic relevance. Nat. Rev. Cancer 5, 263–274 (2005).

  30. 30.

    Nagamura-Inoue, T., Mori, Y., Yizhou, Z., Watanabe, N. & Takahashi, T. A. Differential expansion of umbilical cord blood mononuclear cell-derived natural killer cells dependent on the dose of interleukin-15 with Flt3L. Exp. Hematol. 32, 202–209 (2004).

  31. 31.

    Lin, S. J., Yang, M. H., Chao, H. C., Kuo, M. L. & Huang, J. L. Effect of interleukin-15 and Flt3-ligand on natural killer cell expansion and activation: umbilical cord vs. adult peripheral blood mononuclear cells. Pediatr. Allergy Immunol. 11, 168–174 (2000).

  32. 32.

    Stanietsky, N. et al. Mouse TIGIT inhibits NK-cell cytotoxicity upon interaction with PVR. Eur. J. Immunol. 43, 2138–2150 (2013).

  33. 33.

    Gur, C. et al. Binding of the Fap2 protein of Fusobacterium nucleatum to human inhibitory receptor TIGIT protects tumors from immune cell attack. Immunity 42, 344–355 (2015).

  34. 34.

    Segal, N. H. et al. A phase I dose escalation and cohort expansion study of lirilumab (anti-KIR; BMS-986015) in combination with nivolumab (anti-PD-1; BMS-936558, ONO-4538) in advanced solid tumors. ASCO Annual Meeting Proc. 2014, TPS3115 (2014).

  35. 35.

    Zheng, M., Sun, R., Wei, H. & Tian, Z. NK cells help induce anti-hepatitis B virus CD8+ T cell immunity in mice. J. Immunol. 196, 4122–4131 (2016).

  36. 36.

    Liu, Y. et al. Uncompromised NK cell activation is essential for virus-specific CTL activity during acute influenza virus infection. Cell. Mol. Immunol. (2017).

  37. 37.

    Twyman-Saint Victor, C. et al. Radiation and dual checkpoint blockade activate non-redundant immune mechanisms in cancer. Nature 520, 373–377 (2015).

  38. 38.

    Yang, X. et al. Targeting the tumor microenvironment with interferon-β bridges innate and adaptive immune responses. Cancer Cell 25, 37–48 (2014).

  39. 39.

    Hou, X. et al. CD205-TLR9-IL-12 axis contributes to CpG-induced oversensitive liver injury in HBsAg transgenic mice by promoting the interaction of NKT cells with Kupffer cells. Cell. Mol. Immunol. 14, 675–684 (2017).

Download references


We acknowledged funding support from the following sources: Natural Science Foundation of China (#81788101 to Z.T., #31570893 to R.S., and #81501355 to J.B.); Chinese Academy of Science (XDPB030301 to Z.T.); the Ministry of Science & Technology of China (2017ZX10203206003 to R.S., 2017ZX10202203-002-001 to Y.C.); the Science and Technology Innovation Fund of Shenzhen (JCYJ20150521094519472 and JCYJ20150630114942288 to J.B.). We thank Bristol-Myers Squibb for Tigit–/–mice; T. W. Mak (University of Toronto) for the Nfil3+/– mice; S. Su (Shantou University) for GKO mice; L. Bai and Z. Lian (University of Science and Technology of China) for C57BL/6 J Cd4–/– mice; X. Wang (Inner Mongolia University) for Rag2–/– mice; E. Vivier (INSERM) for Ncr1iCre/+ mice; and Z. Fan (Institute of Biophysics, Chinese Academy of Sciences) for Tigitfl/fl mice.

Author information

Q.Z., R.S. and Z.T. initiated and designed the research; Q.Z. performed all the experiments and analyzed and interpreted results; Q.Z., J.B. and Z.T. wrote the manuscript; X.Z., Y.C., H.P. and H. Wei contributed to discussions of results; and H. Wang, W.W., Z.W. and Q.W. provided clinical specimens, and clinical and pathological information.

Competing interests

The authors declare no competing interests.

Correspondence to Rui Sun or Zhigang Tian.

Integrated supplementary information

  1. Supplementary Figure 1 Phenotypes of tumor-infiltrating lymphocytes.

    Supplementary Figure 1. Phenotypes of tumor-infiltrating lymphocytes. (a) Frequency and MFI of TIGIT+ cells in CD8+ T cells (CD45+CD3+CD56) from peritumoral regions (PT) and intratumoral regions (IT) in patients with CRC (n = 16). (b) Representative flow cytometry plot of TIGIT expression in NK cells from the spleen, liver and lung as described in Fig. 1b. (c) Frequencies of TIGIT+ in CD8+ T cells in B16-pulmonary metastasis model (n = 4 mice per group). (d) Representative flow cytometry plots of tumor-infiltrating NK cells and TIGIT expression on NK cells in four subcutaneous tumor models. (e) Frequencies of TIGIT+ cells in CD8+ T cells as described in Fig. 1c (n = 5 mice per group). (f, g) Frequencies of PD-1+, or CTLA-4+ cells in CD8+ or CD4+ T cells as described in Fig. 1d (n = 5 mice per group). (h) Frequencies of TIGIT+CTLA-4, TIGIT+CTLA-4+, and TIGITCTLA-4+ cells in tumor-infiltrating CD4+ T cells (n = 5 mice per group). (i) Frequencies of CD226, CD96, LAG3, TIM3, Ly49A, Ly49C/I, NKG2A and NKG2D (n = 16, 16, 15, 23, 5, 5, 5, 5 mice) in TIGIT and TIGIT+ tumor-infiltrating NK cells in B16-pulmonary metastasis model. (b-i) Data are representative of at least three independent experiments. ns, not significant (P > 0.05), *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; Error bars are mean ± s. e. m. (c, e, f, g); paired two-tailed t-test (a, i); one-way ANOVA followed by Tukey's (c, h) or Dunnett's (e, f, g) multiple comparisons test.

  2. Supplementary Figure 2 The TIGIT ligand CD155 is abundant in human and murine tumors.

    (a) Representative images of immunolabeling for CD155 and CD112 in human tumor tissues (paraffin sections from 20 tumor patients with colorectal cancer or lung cancer). Scale bars are 100 μm. (b) The expression of CD155 and CD112 on tumor cells we used in the murine tumor models. (a, b) The experiment was repeated independently for four times with similar results.

  3. Supplementary Figure 3 Conditional knockout of TIGIT on NK cells prevents the exhaustion of TILs.

    (a) Representative histograms of TIGIT expression on NK and T cells from Tigitfl/fl- Ncr1iCre/+, Tigitfl/fl- Ncr1+/+, Tigit–/– and WT mice. Grey shadow represent isotype staining. The experiment was repeated independently for three times with similar results. (b, c) Tigitfl/fl- Ncr1iCre/+ and Tigitfl/fl- Ncr1+/+ mice were intravenously injected with 1.5 × 105 B16/F10 cells (n = 4). TILs were analyzed 15-d post challenge. (b) Expression of CD226 and CD96 in tumor-infiltrating NK cells. (c) Expression of PD-1 and Tim3 in tumor-infiltrating CD8+ T cells. (b-c) Data are representative of three independent experiments. ns, not significant (P > 0.05), *P < 0.05, **P < 0.01; Error bars are mean ± s. e. m. (b, c); unpaired two-tailed t-test (b, c).

  4. Supplementary Figure 4 TIGIT blockade alone inhibits tumor growth and reverses the exhaustion of tumor-infiltrating CD8+ T cells.

    (a) Expression of CD107a, TNF, and IFN-γ in tumor-infiltrating CD8+ T cells in CT26-tumor bearing mice (n = 5 mice per group). (b, c) B16 melanoma model. Mice were subcutaneously injected with 5 × 104 B16/F10 cells, and treated with anti-TIGIT mAb and control IgG for 3 times (once every 3 d). (b) Tumor size over time (anti-TIGIT, IgG, PBS, n = 6, 5, 4 mice). (c) Survival of mice (anti-TIGIT, IgG, PBS, n = 7, 7, 6 mice). Data are representative of three (a) or two (b, c) independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001; Error bars are mean ± s. e. m. (a, b); unpaired two-tailed t-test (a); multiple t tests (b); Mantel-Cox test (c).

  5. Supplementary Figure 5 pLIVE-mFlt3L injection accumulates DCs and NK cells, and promotes the activation of NK cells.

    (a-c), Mice were hydro-dynamically injected with 10 μg pLIVE-mFlt3L or pLIVE-NULL. Serum was collected at indicated time post injection. The peripheral blood was collected from the tail vein 2 weeks after the injection. Splenic and Hepatic cells were isolated 4 weeks after the injection. (a) Soluble Flt3L level in serum (n = 4 mice per group) and the frequency of DCs in PBMC (pLIVE-NULL, pLIVE-Flt3L, n = 4, 7 mice). (b) The frequency and absolute numbers of DCs cells in spleen and liver (n = 3 mice per group). (c) The frequency and absolute numbers of NK cells and CD69+ NK cells in spleen and liver (n = 5 mice per group). (d) The frequency of NK cells, CD69+ NK cells and absolute number of CD69+ NK cells in lung from B16 pulmonary metastasis model. Mice were treated as described in Fig. 3n (n = 4 mice per group). (a-d) Data are representative of three independent experiments. ns, not significant (P > 0.05), *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; Error bars are mean ± s. e. m. (a-d); Two-way ANOVA followed by Sidak's multiple comparisons test (a left); unpaired two-tailed t-test (a right, b, c); one-way ANOVA followed by Tukey's multiple comparisons test (d).

  6. Supplementary Figure 6 TIGIT blockade inhibits tumor growth in Rag2–/–and NOD-SCID mice.

    (a, c) Tumor size over time (Rag2–/–, IgG, anti-TIGIT, n = 6, 6; NOD-SCID, IgG, anti-TIGIT, n = 16, 18). (b, d) Expression of IFN-γ (n = 5 mice per group), TNF (n = 5 mice per group) and CD226 (n = 6 mice per group) in tumor-infiltrating NK cells. (a-d) Data are representative of two independent experiments. ns, not significant (P > 0.05), *P < 0.05, **P < 0.01; Error bars are mean ±s. e. m. (a-d); multiple t tests (a, c); unpaired two-tailed t-test (b, d).

Supplementary information

  1. Supplementary Figures and Text

    Supplementary Figures 1-6 and Supplementary Table 1

  2. Reporting Summary

Rights and permissions

To obtain permission to re-use content from this article visit RightsLink.

About this article

Further reading

Fig. 1: TIGIT acts as a newly identified exhaustion marker for tumor infiltrating NK cells.
Fig. 2: TIGIT deficiency inhibits tumor growth and prevents the exhaustion of TILs.
Fig. 3: Blockade of TIGIT inhibits tumor growth and prevents exhaustion of tumor-infiltrating NK cells.
Fig. 4: Blockade of TIGIT has a protective role in mice with adaptive immunodeficiency.
Fig. 5: NK cells are critical for the anti-tumor efficacy of blockade of TIGIT.
Fig. 6: Blockade of TIGIT improves memory response to tumor re-challenge.
Supplementary Figure 1: Phenotypes of tumor-infiltrating lymphocytes.
Supplementary Figure 2: The TIGIT ligand CD155 is abundant in human and murine tumors.
Supplementary Figure 3: Conditional knockout of TIGIT on NK cells prevents the exhaustion of TILs.
Supplementary Figure 4: TIGIT blockade alone inhibits tumor growth and reverses the exhaustion of tumor-infiltrating CD8+ T cells.
Supplementary Figure 5: pLIVE-mFlt3L injection accumulates DCs and NK cells, and promotes the activation of NK cells.
Supplementary Figure 6: TIGIT blockade inhibits tumor growth in Rag2–/–and NOD-SCID mice.