Skip to main content

Thank you for visiting 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.

Inhibition of the dipeptidyl peptidase DPP4 (CD26) reveals IL-33-dependent eosinophil-mediated control of tumor growth


Post-translational modification of chemokines mediated by the dipeptidyl peptidase DPP4 (CD26) has been shown to negatively regulate lymphocyte trafficking, and its inhibition enhances T cell migration and tumor immunity by preserving functional chemokine CXCL10. By extending those initial findings to pre-clinical models of hepatocellular carcinoma and breast cancer, we discovered a distinct mechanism by which inhibition of DPP4 improves anti-tumor responses. Administration of the DPP4 inhibitor sitagliptin resulted in higher concentrations of the chemokine CCL11 and increased migration of eosinophils into solid tumors. Enhanced tumor control was preserved in mice lacking lymphocytes and was ablated after depletion of eosinophils or treatment with degranulation inhibitors. We further demonstrated that tumor-cell expression of the alarmin IL-33 was necessary and sufficient for eosinophil-mediated anti-tumor responses and that this mechanism contributed to the efficacy of checkpoint-inhibitor therapy. These findings provide insight into IL-33- and eosinophil-mediated tumor control, revealed when endogenous mechanisms of DPP4 immunoregulation are inhibited.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: T cells are not required for the DPP4i-mediated anti-tumor effect in HCC and breast cancer syngeneic models.
Fig. 2: DPP4i enhances eosinophilia in solid tumors.
Fig. 3: Eosinophils are required for DPP4i-mediated anti-tumor response.
Fig. 4: DPP4i enhances anti-tumor responses via a CCL11-mediated mechanism.
Fig. 5: IL-33 expression promotes eosinophil-mediated anti-tumor responses in the presence of DPP4i.
Fig. 6: Type 1 immunity and type 2 immunity collaborate to achieve tumor immunotherapy.

Data availability

The raw data that support the findings of this study are available from the corresponding author upon request.


  1. 1.

    Speiser, D. E., Ho, P.-C. & Verdeil, G. Regulatory circuits of T cell function in cancer. Nat. Rev. Immunol. 16, 599–611 (2016).

    CAS  Article  Google Scholar 

  2. 2.

    Proost, P. et al. Amino-terminal truncation of CXCR3 agonists impairs receptor signaling and lymphocyte chemotaxis, while preserving antiangiogenic properties. Blood 98, 3554–3561 (2001).

    CAS  Article  Google Scholar 

  3. 3.

    Vanheule, V., Metzemaekers, M., Janssens, R., Struyf, S. & Proost, P. How post-translational modifications influence the biological activity of chemokines. Cytokine 109, 29–51 (2018).

    CAS  Article  Google Scholar 

  4. 4.

    Casrouge, A. et al. Evidence for an antagonist form of the chemokine CXCL10 in patients chronically infected with HCV. J. Clin. Invest. 121, 308–317 (2011).

    CAS  Article  Google Scholar 

  5. 5.

    Cho, S. Y. et al. The effect of CXCL12 processing on CD34+ cell migration in myeloproliferative neoplasms. Cancer Res. 70, 3402–3410 (2010).

    CAS  Article  Google Scholar 

  6. 6.

    Barreira da Silva, R. et al. Dipeptidylpeptidase 4 inhibition enhances lymphocyte trafficking, improving both naturally occurring tumor immunity and immunotherapy. Nat. Immunol. 16, 850–858 (2015).

    CAS  Article  Google Scholar 

  7. 7.

    Lam, C. S.-C. et al. Prognostic significance of CD26 in patients with colorectal cancer. PLoS ONE 9, e98582 (2014).

    Article  Google Scholar 

  8. 8.

    Liang, P.-I. et al. DPP4/CD26 overexpression in urothelial carcinoma confers an independent prognostic impact and correlates with intrinsic biological aggressiveness. Oncotarget 8, 2995–3008 (2017).

    PubMed  Google Scholar 

  9. 9.

    Yamaguchi, U. et al. Distinct gene expression-defined classes of gastrointestinal stromal tumor. J. Clin. Oncol. 26, 4100–4108 (2008).

    CAS  Article  Google Scholar 

  10. 10.

    Carretero, R. et al. Eosinophils orchestrate cancer rejection by normalizing tumor vessels and enhancing infiltration of CD8+ T cells. Nat. Immunol. 16, 609–617 (2015).

    CAS  Article  Google Scholar 

  11. 11.

    Munitz, A. & Levi-Schaffer, F. Eosinophils: ‘new’ roles for ‘old’ cells. Allergy. 59, 268–275 (2004).

    CAS  Article  Google Scholar 

  12. 12.

    Kubo, H., Loegering, D. A., Adolphson, C. R. & Gleich, G. J. Cytotoxic properties of eosinophil granule major basic protein for tumor cells. Int. Arch. Allergy Immunol. 118, 426–428 (1999).

    CAS  Article  Google Scholar 

  13. 13.

    Zhang, H. & Verkman, A. S. Eosinophil pathogenicity mechanisms and therapeutics in neuromyelitis optica. J. Clin. Invest. 123, 2306–2316 (2013).

    CAS  Article  Google Scholar 

  14. 14.

    Krishna, M. C. et al. Do nitroxide antioxidants act as scavengers of O2 . or as SOD mimics? J. Biol. Chem. 271, 26026–26031 (1996).

    CAS  Article  Google Scholar 

  15. 15.

    Rothenberg, M. E. & Hogan, S. P. The eosinophil. Annu. Rev. Immunol. 24, 147–174 (2006).

    CAS  Article  Google Scholar 

  16. 16.

    Gatault, S., Legrand, F., Delbeke, M., Loiseau, S. & Capron, M. Involvement of eosinophils in the anti-tumor response. Cancer Immunol. Immunother. 61, 1527–1534 (2012).

    Article  Google Scholar 

  17. 17.

    Struyf, S. et al. CD26/dipeptidyl-peptidase IV down-regulates the eosinophil chemotactic potency, but not the anti-HIV activity of human eotaxin by affecting its interaction with CC chemokine receptor 3. J. Immunol. 162, 4903–4909 (1999).

    CAS  PubMed  Google Scholar 

  18. 18.

    Proost, P. et al. Amino-terminal truncation of chemokines by CD26/dipeptidyl-peptidase IV. Conversion of RANTES into a potent inhibitor of monocyte chemotaxis and HIV-1-infection. J. Biol. Chem. 273, 7222–7227 (1998).

    CAS  Article  Google Scholar 

  19. 19.

    Forssmann, U. et al. Inhibition of CD26/dipeptidyl peptidase IV enhances CCL11/eotaxin-mediated recruitment of eosinophils in vivo. J. Immunol. 181, 1120–1127 (2008).

    CAS  Article  Google Scholar 

  20. 20.

    Wasmer, M.-H. & Krebs, P. The role of IL-33-dependent inflammation in the tumor microenvironment. Front. Immunol. 7, 682 (2016).

    PubMed  Google Scholar 

  21. 21.

    Johnston, L. K. et al. IL-33 precedes IL-5 in regulating eosinophil commitment and is required for eosinophil homeostasis. J. Immunol. 197, 3445–3453 (2016).

    CAS  Article  Google Scholar 

  22. 22.

    Gao, X. et al. Tumoral expression of IL-33 inhibits tumor growth and modifies the tumor microenvironment through CD8+ T and NK cells. J. Immunol. 194, 438–445 (2015).

    CAS  Article  Google Scholar 

  23. 23.

    Tepper, R. I., Coffman, R. L. & Leder, P. An eosinophil-dependent mechanism for the antitumor effect of interleukin-4. Science 257, 548–551 (1992).

    CAS  Article  Google Scholar 

  24. 24.

    Mattes, J. et al. Immunotherapy of cytotoxic T cell-resistant tumors by T helper 2 cells: an eotaxin and STAT6-dependent process. J. Exp. Med. 197, 387–393 (2003).

    CAS  Article  Google Scholar 

  25. 25.

    Lucarini, V. et al. IL-33 restricts tumor growth and inhibits pulmonary metastasis in melanoma-bearing mice through eosinophils. Oncoimmunology 6, e1317420 (2017).

    Article  Google Scholar 

  26. 26.

    Bousquet, J. et al. Eosinophilic inflammation in asthma. N. Engl. J. Med. 323, 1033–1039 (1990).

    CAS  Article  Google Scholar 

  27. 27.

    Ma, W. et al. CCR3 is essential for skin eosinophilia and airway hyperresponsiveness in a murine model of allergic skin inflammation. J. Clin. Invest. 109, 621–628 (2002).

    CAS  Article  Google Scholar 

  28. 28.

    Crapster-Pregont, M., Yeo, J., Sanchez, R. L. & Kuperman, D. A. Dendritic cells and alveolar macrophages mediate IL-13-induced airway inflammation and chemokine production. J. Allergy Clin. Immunol. 129, 1621–7.e3 (2012).

    CAS  Article  Google Scholar 

  29. 29.

    Wills-Karp, M. et al. Interleukin-13: central mediator of allergic asthma. Science 282, 2258–2261 (1998).

    CAS  Article  Google Scholar 

  30. 30.

    Corren, J. et al. Lebrikizumab treatment in adults with asthma. N. Engl. J. Med. 365, 1088–1098 (2011).

    CAS  Article  Google Scholar 

  31. 31.

    Wenzel, S., Wilbraham, D., Fuller, R., Getz, E. B. & Longphre, M. Effect of an interleukin-4 variant on late phase asthmatic response to allergen challenge in asthmatic patients: results of two phase 2a studies. Lancet 370, 1422–1431 (2007).

    CAS  Article  Google Scholar 

  32. 32.

    Shiobara, T. et al. Dipeptidyl peptidase-4 is highly expressed in bronchial epithelial cells of untreated asthma and it increases cell proliferation along with fibronectin production in airway constitutive cells. Respir. Res. 17, 28 (2016).

    Article  Google Scholar 

  33. 33.

    Zhen, G. et al. IL-13 and epidermal growth factor receptor have critical but distinct roles in epithelial cell mucin production. Am. J. Respir. Cell Mol. Biol. 36, 244–253 (2007).

    CAS  Article  Google Scholar 

  34. 34.

    Brightling, C. E. et al. Efficacy and safety of tralokinumab in patients with severe uncontrolled asthma: a randomised, double-blind, placebo-controlled, phase 2b trial. Lancet Respir. Med. 3, 692–701 (2015).

    CAS  Article  Google Scholar 

  35. 35.

    Samoszuk, M. et al. Increased blood clotting, microvascular density, and inflammation in eotaxin-secreting tumors implanted into mice. Am. J. Pathol. 165, 449–456 (2004).

    Article  Google Scholar 

  36. 36.

    Reichman, H., Karo-Atar, D. & Munitz, A. Emerging roles for eosinophils in the tumor microenvironment. Trends Cancer 2, 664–675 (2016).

    Article  Google Scholar 

  37. 37.

    Dorta, R. G. et al. Tumour-associated tissue eosinophilia as a prognostic factor in oral squamous cell carcinomas. Histopathology 41, 152–157 (2002).

    CAS  Article  Google Scholar 

  38. 38.

    Fernández-Aceñero, M. J., Galindo-Gallego, M., Sanz, J. & Aljama, A. Prognostic influence of tumor-associated eosinophilic infiltrate in colorectal carcinoma. Cancer 88, 1544–1548 (2000).

    Article  Google Scholar 

  39. 39.

    Costello, R., O’Callaghan, T. & Sébahoun, G. Eosinophils and antitumour response. Rev. Med. Interne. 26, 479–484 (2005).

    CAS  Article  Google Scholar 

  40. 40.

    Caruso, R. A. et al. Ultrastructural descriptions of heterotypic aggregation between eosinophils and tumor cells in human gastric carcinomas. Ultrastruct. Pathol. 35, 145–149 (2011).

    Article  Google Scholar 

  41. 41.

    Stenfeldt, A.-L. & Wennerås, C. Danger signals derived from stressed and necrotic epithelial cells activate human eosinophils. Immunology 112, 605–614 (2004).

    CAS  Article  Google Scholar 

  42. 42.

    Griesenauer, B. & Paczesny, S. The ST2/IL-33 axis in immune cells during inflammatory diseases. Front. Immunol. 8, 475 (2017).

    Article  Google Scholar 

  43. 43.

    Bourgeois, E. et al. The pro-Th2 cytokine IL-33 directly interacts with invariant NKT and NK cells to induce IFN-γ production. Eur. J. Immunol. 39, 1046–1055 (2009).

    CAS  Article  Google Scholar 

  44. 44.

    Bonilla, W. V. et al. The alarmin interleukin-33 drives protective antiviral CD8+ T cell responses. Science 335, 984–989 (2012).

    CAS  Article  Google Scholar 

  45. 45.

    Oboki, K., Nakae, S., Matsumoto, K. & Saito, H. IL-33 and airway inflammation. Allergy Asthma Immunol. Res. 3, 81–88 (2011).

    CAS  Article  Google Scholar 

  46. 46.

    Vasanthakumar, A. et al. The transcriptional regulators IRF4, BATF and IL-33 orchestrate development and maintenance of adipose tissue-resident regulatory T cells. Nat. Immunol. 16, 276–285 (2015).

    CAS  Article  Google Scholar 

  47. 47.

    Lu, B., Yang, M. & Wang, Q. Interleukin-33 in tumorigenesis, tumor immune evasion, and cancer immunotherapy. J. Mol. Med. 94, 535–543 (2016).

    CAS  Article  Google Scholar 

  48. 48.

    Kim, J. et al. Intratumorally establishing type 2 innate lymphoid cells blocks tumor growth. J. Immunol. 196, 2410–2423 (2016).

    CAS  Article  Google Scholar 

  49. 49.

    Jovanovic, I. et al. ST2 deletion enhances innate and acquired immunity to murine mammary carcinoma. Eur. J. Immunol. 41, 1902–1912 (2011).

    CAS  Article  Google Scholar 

  50. 50.

    Xiao, P. et al. Interleukin 33 in tumor microenvironment is crucial for the accumulation and function of myeloid-derived suppressor cells. Oncoimmunology 5, e1063772 (2016).

    Article  Google Scholar 

Download references


Funding for the work was provided by Fondation ARC pour la recherche sur le cancer, Institut national de la santé et de la recherche médicale (Inserm), Fondation pour la recherche médicale (FRM, FDM 40432) and LabEx Immuno-Onco (ANR). Thanks to H. Saklani, M. A. Ingersoll and T. Canton for their help with mouse experimental work and ethical statement.

Author information




C.H., S.P., R.B.d.S. and M.L.A. designed the study. C.H. and R.B.d.S. designed, carried out and analyzed experiments. J.B., T.N., D.D., V.M., J.M.S. and G.E. provided technical and intellectual assistance. V.B., W.P. and W.S. conducted mass spectrometry experiments. B.L. provided tumor cell lines. V.P. and J.Z. conducted histological experiments. C.H., R.B.d.S. and M.L.A. wrote the manuscript. M.L.A., R.B.d.S. and S.P. supervised the study.

Corresponding author

Correspondence to Matthew L. Albert.

Ethics declarations

Competing interests

R.B.d.S., M.L.A., J.Z., T.N., W.P., W.S. and J.M.S. are current employees of Genentech, a member of the Roche group.

Additional information

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

Integrated supplementary information

Supplementary Figure 1 DPP4i diminishes tumor volume and DPP enzymatic activity in Hepa1-6 and EMT6 subcutaneous tumors.

(a, b) WT mice were fed with DPP4i or with Ctrl chow and subcutaneously injected with Hepa 1-6 (a) or EMT6 (b) cells. Tumors were collected at day 10 post-inoculation and tumor mass was determined. Bars represent median values (n = 17 (Ctrl, panel a), 18 (DPP4i, panel a) or 6 per group (b)). (c, d) Eight days after Hepa 1-6 (c) or EMT6 (d) tumor cell inoculation, tumors were collected and homogenates were prepared. DPP4 activity per tumor mass was determined and normalized to the amount of DPP4 protein (RLU, relative luminescence units). Bars represent median values (n = 4 per group (c), 6 (Ctrl, d) or 7 (DPP4i, d) mice). (e) Hepa 1-6 cells were cultured in the presence of 5 (C1), 0.05 (C2) or 0.005 (C3) μg/ml of DPP4i, or left untreated (). Cell confluence was measured over time using intra-incubator microscopy (mean ± SEM, n = 3 technical replicates). Each dot corresponds to one mouse. Data are representative of two independent experiments (be) or pooled from two (a) independent experiments. NS, not significant; *P < 0.05, ****P < 0.0001. Significance was determined using two-sided Mann–Whitney test (ad). (Allergy 63, 1156–1163, 2008)

Supplementary Figure 2 DPP4i enhances infiltration of eosinophils in the EMT6 breast model and enrichment for T cells in the parenchyma of Hepa1-6 subcutaneous tumors.

(a, b) Gating strategy for identification of tumor infiltrating leukocytes. (c, d) WT mice were fed with DPP4i or with Ctrl chow and subcutaneously injected with EMT6 cells. Tumors were collected at day 10 post-inoculation and tumor-associated leukocytes were analyzed by flow cytometry. Number of tumor infiltrating T cells and eosinophils is shown. Each dot corresponds to one mouse (n = 11 (Ctrl, c) or 12 (DPP4i, c) or 12 per group (d)). Bars represent median values. (e, f) WT mice were treated as in c and inoculated with Hepa1-6 cells. Representative images (scale, 40 μm) and quantification of CD3 expressing cells in Hepa 1-6 tumors collected 10 d after inoculation. Each dot corresponds to one mouse (n = 8 (Ctrl) or 7 (DPP4i) mice). Bars represent median values. (g) Rag2–/–γc–/– mice fed with Ctrl or DPP4i were inoculated with Hepa 1-6 cells. Tumor volumes were measured over time (mean ± SEM, n = 4 mice (Ctrl) or 5 mice (DPP4i). Data are pooled from two (c, d) independent experiments. Histological analysis on T cell distribution and tumor growth on Rag2–/–γc–/– was done once. NS, not significant; *P < 0.05, **P < 0.01. Significance was determined using or two-sided Mann–Whitney test (c, d, f) or two-way analysis of variance (g).

Supplementary Figure 3 DPP4 does not truncate mouse CCL24 but reduces CCL11-mediated internalization of CCR3 in eosinophils.

(a) Recombinant mCCL24 was incubated in the presence or absence of mDPP4 and analyzed by mass spectrometry. Numbers indicate the molecular weight (in Daltons). (b) WT mice fed with Ctrl or with DPP4i were injected intravenously with 1 μg of CCL11 and blood was collected 1 h after injection. Representative histogram of CCR3 expression on eosinophils is shown. Quantification was determined by flow cytometry. Each dot represents one mouse (n = 7 (Ctrl) or 8 (DPP4i) mice). Data are representative of two independent experiments yielding similar results. Bars represent median values. NS, not significant; **P < 0.01. Significance was determined using or two-sided Mann–Whitney test.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Hollande, C., Boussier, J., Ziai, J. et al. Inhibition of the dipeptidyl peptidase DPP4 (CD26) reveals IL-33-dependent eosinophil-mediated control of tumor growth. Nat Immunol 20, 257–264 (2019).

Download citation

Further reading


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