Abstract
Macrophage phagocytosis of tumor cells mediated by CD47-specific blocking antibodies has been proposed to be the major effector mechanism in xenograft models. Here, using syngeneic immunocompetent mouse tumor models, we reveal that the therapeutic effects of CD47 blockade depend on dendritic cell but not macrophage cross-priming of T cell responses. The therapeutic effects of anti-CD47 antibody therapy were abrogated in T cell–deficient mice. In addition, the antitumor effects of CD47 blockade required expression of the cytosolic DNA sensor STING, but neither MyD88 nor TRIF, in CD11c+ cells, suggesting that cytosolic sensing of DNA from tumor cells is enhanced by anti-CD47 treatment, further bridging the innate and adaptive responses. Notably, the timing of administration of standard chemotherapy markedly impacted the induction of antitumor T cell responses by CD47 blockade. Together, our findings indicate that CD47 blockade drives T cell–mediated elimination of immunogenic tumors.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Gardai, S.J. et al. Cell-surface calreticulin initiates clearance of viable or apoptotic cells through trans-activation of LRP on the phagocyte. Cell 123, 321–334 (2005).
Chao, M.P. et al. Calreticulin is the dominant pro-phagocytic signal on multiple human cancers and is counterbalanced by CD47. Sci. Transl. Med. 2, 63ra94 (2010).
Chao, M.P., Majeti, R. & Weissman, I.L. Programmed cell removal: a new obstacle in the road to developing cancer. Nat. Rev. Cancer 12, 58–67 (2012).
Oldenborg, P.A. et al. Role of CD47 as a marker of self on red blood cells. Science 288, 2051–2054 (2000).
Blazar, B.R. et al. CD47 (integrin-associated protein) engagement of dendritic cell and macrophage counterreceptors is required to prevent the clearance of donor lymphohematopoietic cells. J. Exp. Med. 194, 541–549 (2001).
Barclay, A.N. & Van den Berg, T.K. The interaction between signal regulatory protein alpha (SIRPα) and CD47: structure, function, and therapeutic target. Annu. Rev. Immunol. 32, 25–50 (2014).
Yamao, T. et al. Negative regulation of platelet clearance and of the macrophage phagocytic response by the transmembrane glycoprotein SHPS-1. J. Biol. Chem. 277, 39833–39839 (2002).
Olsson, M., Bruhns, P., Frazier, W.A., Ravetch, J.V. & Oldenborg, P.A. Platelet homeostasis is regulated by platelet expression of CD47 under normal conditions and in passive immune thrombocytopenia. Blood 105, 3577–3582 (2005).
Tsai, R.K. & Discher, D.E. Inhibition of “self” engulfment through deactivation of myosin-II at the phagocytic synapse between human cells. J. Cell Biol. 180, 989–1003 (2008).
Poels, L.G. et al. Monoclonal antibody against human ovarian tumor-associated antigens. J. Natl. Cancer Inst. 76, 781–791 (1986).
Jaiswal, S. et al. CD47 is upregulated on circulating hematopoietic stem cells and leukemia cells to avoid phagocytosis. Cell 138, 271–285 (2009).
Majeti, R. et al. CD47 is an adverse prognostic factor and therapeutic antibody target on human acute myeloid leukemia stem cells. Cell 138, 286–299 (2009).
Rendtlew Danielsen, J.M., Knudsen, L.M., Dahl, I.M., Lodahl, M. & Rasmussen, T. Dysregulation of CD47 and the ligands thrombospondin 1 and 2 in multiple myeloma. Br. J. Haematol. 138, 756–760 (2007).
Chan, K.S. et al. Identification, molecular characterization, clinical prognosis, and therapeutic targeting of human bladder tumor-initiating cells. Proc. Natl. Acad. Sci. USA 106, 14016–14021 (2009).
Chan, K.S., Volkmer, J.P. & Weissman, I. Cancer stem cells in bladder cancer: a revisited and evolving concept. Curr. Opin. Urol. 20, 393–397 (2010).
Chao, M.P. et al. Anti-CD47 antibody synergizes with rituximab to promote phagocytosis and eradicate non-Hodgkin lymphoma. Cell 142, 699–713 (2010).
Willingham, S.B. et al. The CD47-signal regulatory protein α (SIRPa) interaction is a therapeutic target for human solid tumors. Proc. Natl. Acad. Sci. USA 109, 6662–6667 (2012).
Chao, M.P. et al. Therapeutic antibody targeting of CD47 eliminates human acute lymphoblastic leukemia. Cancer Res. 71, 1374–1384 (2011).
Chao, M.P. et al. Extranodal dissemination of non-Hodgkin lymphoma requires CD47 and is inhibited by anti-CD47 antibody therapy. Blood 118, 4890–4901 (2011).
Soto-Pantoja, D.R. et al. CD47 in the tumor microenvironment limits cooperation between antitumor T-cell immunity and radiotherapy. Cancer Res. 74, 6771–6783 (2014).
Lee, Y. et al. Therapeutic effects of ablative radiation on local tumor require CD8+ T cells: changing strategies for cancer treatment. Blood 114, 589–595 (2009).
Brown, E.J. & Frazier, W.A. Integrin-associated protein (CD47) and its ligands. Trends Cell Biol. 11, 130–135 (2001).
Weiskopf, K. et al. Engineered SIRPα variants as immunotherapeutic adjuvants to anticancer antibodies. Science 341, 88–91 (2013).
Tseng, D. et al. Anti-CD47 antibody-mediated phagocytosis of cancer by macrophages primes an effective antitumor T-cell response. Proc. Natl. Acad. Sci. USA 110, 11103–11108 (2013).
Burnette, B.C. et al. The efficacy of radiotherapy relies upon induction of type I interferon-dependent innate and adaptive immunity. Cancer Res. 71, 2488–2496 (2011).
Diamond, M.S. et al. Type I interferon is selectively required by dendritic cells for immune rejection of tumors. J. Exp. Med. 208, 1989–2003 (2011).
Fuertes, M.B. et al. Host type I IFN signals are required for antitumor CD8+ T cell responses through CD8α+ dendritic cells. J. Exp. Med. 208, 2005–2016 (2011).
Stagg, J. et al. Anti-ErbB-2 mAb therapy requires type I and II interferons and synergizes with anti-PD-1 or anti-CD137 mAb therapy. Proc. Natl. Acad. Sci. USA 108, 7142–7147 (2011).
Yang, X. et al. Targeting the tumor microenvironment with interferon-β bridges innate and adaptive immune responses. Cancer Cell 25, 37–48 (2014).
Chen, G.Y. & Nunez, G. Sterile inflammation: sensing and reacting to damage. Nat. Rev. Immunol. 10, 826–837 (2010).
Desmet, C.J. & Ishii, K.J. Nucleic acid sensing at the interface between innate and adaptive immunity in vaccination. Nat. Rev. Immunol. 12, 479–491 (2012).
Kono, H. & Rock, K.L. How dying cells alert the immune system to danger. Nat. Rev. Immunol. 8, 279–289 (2008).
O'Neill, L.A., Golenbock, D. & Bowie, A.G. The history of Toll-like receptors—redefining innate immunity. Nat. Rev. Immunol. 13, 453–460 (2013).
Ishikawa, H. & Barber, G.N. STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling. Nature 455, 674–678 (2008).
Ishikawa, H., Ma, Z. & Barber, G.N. STING regulates intracellular DNA-mediated, type I interferon-dependent innate immunity. Nature 461, 788–792 (2009).
Li, X.D. et al. Pivotal roles of cGAS-cGAMP signaling in antiviral defense and immune adjuvant effects. Science 341, 1390–1394 (2013).
Sun, L., Wu, J., Du, F., Chen, X. & Chen, Z.J. Cyclic GMP-AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway. Science 339, 786–791 (2013).
Wu, J. et al. Cyclic GMP-AMP is an endogenous second messenger in innate immune signaling by cytosolic DNA. Science 339, 826–830 (2013).
Woo, S.R. et al. STING-dependent cytosolic DNA sensing mediates innate immune recognition of immunogenic tumors. Immunity 41, 830–842 (2014).
Deng, L. et al. STING-dependent cytosolic DNA sensing promotes radiation-induced type I interferon-dependent antitumor immunity in immunogenic tumors. Immunity 41, 843–852 (2014).
Obeid, M. et al. Ecto-calreticulin in immunogenic chemotherapy. Immunol. Rev. 220, 22–34 (2007).
Obeid, M. et al. Calreticulin exposure dictates the immunogenicity of cancer cell death. Nat. Med. 13, 54–61 (2007).
Takenaka, K. et al. Polymorphism in Sirpa modulates engraftment of human hematopoietic stem cells. Nat. Immunol. 8, 1313–1323 (2007).
Yamauchi, T. et al. Polymorphic Sirpa is the genetic determinant for NOD-based mouse lines to achieve efficient human cell engraftment. Blood 121, 1316–1325 (2013).
Matsushita, H. et al. Cancer exome analysis reveals a T-cell-dependent mechanism of cancer immunoediting. Nature 482, 400–404 (2012).
Snyder, A. et al. Genetic basis for clinical response to CTLA-4 blockade in melanoma. N. Engl. J. Med. 371, 2189–2199 (2014).
Ma, Y. et al. Anticancer chemotherapy-induced intratumoral recruitment and differentiation of antigen-presenting cells. Immunity 38, 729–741 (2013).
Ma, Y. et al. CCL2/CCR2-dependent recruitment of functional antigen-presenting cells into tumors upon chemotherapy. Cancer Res. 74, 436–445 (2014).
Rovero, S. et al. DNA vaccination against rat her-2/Neu p185 more effectively inhibits carcinogenesis than transplantable carcinomas in transgenic BALB/c mice. J. Immunol. 165, 5133–5142 (2000).
Oldenborg, P.A., Gresham, H.D. & Lindberg, F.P. CD47-signal regulatory protein α (SIRPα) regulates Fcγ and complement receptor-mediated phagocytosis. J. Exp. Med. 193, 855–862 (2001).
Acknowledgements
We thank R. Schreiber (Washington University, St. Louis) for providing us with anti-IFNAR antibody. Ifnar1fl/fl mice were kindly provided by U. Kalinke from the Institute for Experimental Infection Research. Anti-IFN-γ neutralizing mAb (clone R46A2) was provided by Z. Qin (the Institute of Biophysics, CAS). Some anti-CD47 antibody (clone MIAP301) was provided by W. Frazier (Washington University, St. Louis). This research was in part supported by US National Institutes of Health grants CA141975 and C134563 to Y.-X.F., the National 12.5 major project of China (No. 2012ZX10001006002004) to Y.-X.F. and H.P., Chinese Academy of Sciences grant XDA09030303 and 2012CB910203 to Y.-X.F. and a Dean's Award from Washington University to W.A.F.
Author information
Authors and Affiliations
Contributions
X.L., Y.P., H.X. and M.M.X. performed experiments. L.D., J.K., W.A.F. and H.P. provided reagents. X.L., M.M.X. and Y.-X.F. designed and organized experiments. K.C., J.K. and H.P. edited the manuscript. X.L., M.M.X. and Y.-X.F. wrote the paper. Y.-X.F. guided the work.
Corresponding author
Ethics declarations
Competing interests
W.A.F. is founder and a stockholder of Vasculox, Inc.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–6 and Supplementary Table 1 (PDF 572 kb)
Rights and permissions
About this article
Cite this article
Liu, X., Pu, Y., Cron, K. et al. CD47 blockade triggers T cell–mediated destruction of immunogenic tumors. Nat Med 21, 1209–1215 (2015). https://doi.org/10.1038/nm.3931
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nm.3931
This article is cited by
-
The enhanced antitumor activity of bispecific antibody targeting PD-1/PD-L1 signaling
Cell Communication and Signaling (2024)
-
Targeting HDAC6 improves anti-CD47 immunotherapy
Journal of Experimental & Clinical Cancer Research (2024)
-
A pH-dependent anti-CD47 antibody that selectively targets solid tumors and improves therapeutic efficacy and safety
Journal of Hematology & Oncology (2023)
-
Targeting cGAS/STING signaling-mediated myeloid immune cell dysfunction in TIME
Journal of Biomedical Science (2023)
-
The CAR macrophage cells, a novel generation of chimeric antigen-based approach against solid tumors
Biomarker Research (2023)