Adoptive cell therapy (ACT) with antigen-specific T cells has shown remarkable clinical success; however, approaches to safely and effectively augment T cell function, especially in solid tumors, remain of great interest. Here we describe a strategy to 'backpack' large quantities of supporting protein drugs on T cells by using protein nanogels (NGs) that selectively release these cargos in response to T cell receptor activation. We designed cell surface–conjugated NGs that responded to an increase in T cell surface reduction potential after antigen recognition and limited drug release to sites of antigen encounter, such as the tumor microenvironment. By using NGs that carried an interleukin-15 super-agonist complex, we demonstrated that, relative to systemic administration of free cytokines, NG delivery selectively expanded T cells 16-fold in tumors and allowed at least eightfold higher doses of cytokine to be administered without toxicity. The improved therapeutic window enabled substantially increased tumor clearance by mouse T cell and human chimeric antigen receptor (CAR)-T cell therapy in vivo.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from $8.99

All prices are NET prices.


  1. 1.

    & Adoptive cell transfer as personalized immunotherapy for human cancer. Science 348, 62–68 (2015).

  2. 2.

    & Going viral: chimeric antigen receptor T cell therapy for hematological malignancies. Immunol. Rev. 263, 68–89 (2015).

  3. 3.

    et al. T cell immunotherapy: looking forward. Mol. Ther. 22, 1564–1574 (2014).

  4. 4.

    et al. Rational development and characterization of humanized anti–EGFR variant III chimeric antigen receptor T cells for glioblastoma. Sci. Transl. Med. 7, 275ra22 (2015).

  5. 5.

    et al. IL-15 enhances the in vivo antitumor activity of tumor-reactive CD8+ T cells. Proc. Natl. Acad. Sci. USA 101, 1969–1974 (2004).

  6. 6.

    et al. Transforming growth factor–β receptor blockade augments the effectiveness of adoptive T cell therapy of established solid cancers. Clin. Cancer Res. 14, 3966–3974 (2008).

  7. 7.

    et al. Redistribution, hyperproliferation, activation of natural killer cells and CD8 T cells, and cytokine production during first-in-human clinical trial of recombinant human interleukin-15 in patients with cancer. J. Clin. Oncol. 33, 74–82 (2015).

  8. 8.

    et al. Effects of single-dose interleukin-12 exposure on interleukin-12-associated toxicity and interferon-γ production. Blood 90, 2541–2548 (1997).

  9. 9.

    et al. Tumor-infiltrating lymphocytes genetically engineered with an inducible gene encoding interleukin-12 for the immunotherapy of metastatic melanoma. Clin. Cancer Res. 21, 2278–2288 (2015).

  10. 10.

    et al. Active targeting of chemotherapy to disseminated tumors using nanoparticle-carrying T cells. Sci. Transl. Med. 7, 291ra94 (2015).

  11. 11.

    , , , & Therapeutic cell engineering with surface-conjugated synthetic nanoparticles. Nat. Med. 16, 1035–1041 (2010).

  12. 12.

    , , , & Synapse-directed delivery of immunomodulators using T cell–conjugated nanoparticles. Biomaterials 33, 5776–5787 (2012).

  13. 13.

    , & Thiol-disulfide balance: from the concept of oxidative stress to that of redox regulation. Antioxid. Redox Signal. 7, 964–972 (2005).

  14. 14.

    , & Surface thiols of human lymphocytes and their changes after in vitro and in vivo activation. J. Leukoc. Biol. 60, 611–618 (1996).

  15. 15.

    & Trans-plasma membrane electron transport: a cellular assay for NADH- and NADPH-oxidase based on extracellular, superoxide-mediated reduction of the sulfonated tetrazolium salt WST-1. Protoplasma 205, 74–82 (1998).

  16. 16.

    & Cell-surface NAD(P)H-oxidase: relationship to trans-plasma membrane NADH-oxidoreductase and a potential source of circulating NADH-oxidase. Antioxid. Redox Signal. 2, 277–288 (2000).

  17. 17.

    , & Self-immolative linkers literally bridge disulfide chemistry and the realm of thiol-free drugs. Adv. Healthc. Mater. 4, 1887–1890 (2015).

  18. 18.

    et al. Releasable luciferin–transporter conjugates: tools for the real-time analysis of cellular uptake and release. J. Am. Chem. Soc. 128, 6526–6527 (2006).

  19. 19.

    et al. Rendering protein-based particles transiently insoluble for therapeutic applications. J. Am. Chem. Soc. 134, 8774–8777 (2012).

  20. 20.

    et al. Novel human interleukin-15 agonists. J. Immunol. 183, 3598–3607 (2009).

  21. 21.

    et al. Comparison of the superagonist complex, ALT-803, to IL-15 as cancer immunotherapeutics in animal models. Cancer Immunol. Res. 4, 49–60 (2016).

  22. 22.

    , , & Efficient internalization of IL-2 depends on the distal portion of the cytoplasmic tail of the IL-2R common γ-chain and a lymphoid cell environment. J. Immunol. 165, 2556–2562 (2000).

  23. 23.

    et al. Tumor regression and autoimmunity after reversal of a functionally tolerant state of self-reactive CD8+ T cells. J. Exp. Med. 198, 569–580 (2003).

  24. 24.

    et al. Chimeric antigen receptor T cells for sustained remissions in leukemia. N. Engl. J. Med. 371, 1507–1517 (2014).

  25. 25.

    et al. Adoptive immunotherapy for cancer or viruses. Annu. Rev. Immunol. 32, 189–225 (2014).

  26. 26.

    et al. IL-15 superagonist-mediated immunotoxicity: role of NK cells and IFN-γ. J. Immunol. 195, 2353–2364 (2015).

  27. 27.

    , , & Cyclooxgenase-2 inhibiting perfluoropoly (ethylene glycol) ether theranostic nanoemulsions–in vitro study. PLoS One 8, e55802 (2013).

  28. 28.

    & The spectrin–ankyrin skeleton controls CD45 surface display and interleukin-2 production. Immunity 17, 303–315 (2002).

  29. 29.

    et al. Initiation of T cell signaling by CD45 segregation at 'close contacts'. Nat. Immunol. 17, 574–582 (2016).

  30. 30.

    , , & A supramolecular basis for CD45 tyrosine phosphatase regulation in sustained T cell activation. Proc. Natl. Acad. Sci. USA 97, 10138–10143 (2000).

  31. 31.

    et al. Nanoclusters self-assembled from conformation-stabilized influenza M2e as broadly cross-protective influenza vaccines. Nanomedicine (Lond.) 10, 473–482 (2014).

  32. 32.

    et al. Protein adsorption and cell adhesion on nanoscale bioactive coatings formed from poly(ethylene glycol) and albumin microgels. Biomaterials 29, 4481–4493 (2008).

  33. 33.

    et al. PEG-urokinase nanogels with enhanced stability and controllable bioactivity. Soft Matter 8, 2644–2650 (2012).

  34. 34.

    et al. Oxidative stress in malignant melanoma enhances tumor necrosis factor–α secretion of tumor-associated macrophages that promote cancer cell invasion. Antioxid. Redox Signal. 19, 1337–1355 (2013).

  35. 35.

    , , & Injectable, porous and cell-responsive gelatin cryogels. Biomaterials 35, 2477–2487 (2014).

  36. 36.

    et al. Bioresponsive mesoporous silica nanoparticles for triggered drug release. J. Am. Chem. Soc. 133, 19582–19585 (2011).

  37. 37.

    et al. Folate-targeted pH-responsive calcium zoledronate nanoscale metal-organic frameworks: turning a bone anti-resorptive agent into an anticancer therapeutic. Biomaterials 82, 178–193 (2016).

  38. 38.

    et al. pH-sensitive nanoformulated triptolide as a targeted therapeutic strategy for hepatocellular carcinoma. ACS Nano 8, 8027–8039 (2014).

  39. 39.

    et al. Polymer nanoparticles modified with photo- and pH-dual-responsive polypeptides for enhanced and targeted cancer therapy. Mol. Pharm. 13, 1508–1519 (2016).

  40. 40.

    , , & IL-12 release by engineered T cells expressing chimeric antigen receptors can effectively muster an antigen-independent macrophage response on tumor cells that have shut down tumor antigen expression. Cancer Res. 71, 5697–5706 (2011).

  41. 41.

    et al. Local delivery of interleukin-12 using T cells targeting VEGF receptor 2 eradicates multiple vascularized tumors in mice. Clin. Cancer Res. 18, 1672–1683 (2012).

  42. 42.

    et al. Tumor-targeted T cells modified to secrete IL-12 eradicate systemic tumors without need for prior conditioning. Blood 119, 4133–4141 (2012).

  43. 43.

    et al. Membrane-attached cytokines expressed by mRNA electroporation act as potent T cell adjuvants. J. Immunother. 39, 60–70 (2016).

  44. 44.

    et al. Tumor-specific activation of an EGFR-targeting probody enhances therapeutic index. Sci. Transl. Med. 5, 207ra144 (2013).

  45. 45.

    , , , & Remote control of therapeutic T cells through a small-molecule-gated chimeric receptor. Science 350, aab4077 (2015).

  46. 46.

    , & PD-1- and CTLA-4-based inhibitory chimeric antigen receptors (iCARs) divert off-target immunotherapy responses. Sci. Transl. Med. 5, 215ra172 (2013).

  47. 47.

    , , , & Combinatorial antigen recognition with balanced signaling promotes selective tumor eradication by engineered T cells. Nat. Biotechnol. 31, 71–75 (2013).

  48. 48.

    et al. Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing ERBB2. Mol. Ther. 18, 843–851 (2010).

  49. 49.

    et al. IL-15:IL-15 receptor–α superagonist complex: high-level co-expression in recombinant mammalian cells, purification and characterization. Cytokine 56, 804–810 (2011).

  50. 50.

    et al. In vivo targeting of adoptively transferred T cells with antibody- and cytokine-conjugated liposomes. J. Control. Release 172, 426–435 (2013).

Download references


We thank K.D. Wittrup (MIT) for the gift of the engineered IL-2-Fc constructs and the Koch Institute Swanson Biotechnology Center for technical support on flow cytometry, IVIS imaging and MALDI mass spectrometry. This work was supported in part by the Ragon Institute of MGH, MIT and Harvard (D.J.I.), the Melanoma Research Alliance (award 306833; D.J.I.), the NIH (Koch Institute Support (core) grant P30-CA14051 from the National Cancer Institute and CA172164; D.J.I.) and the Koch Institute Marble Center for Cancer Nanomedicine (D.J.I.). L.T. was funded by a Cancer Research Institute (CRI) Irvington Postdoctoral Fellowship, and Y.Z. was supported by a National Science fellowship from the Agency for Science, Technology and Research, Singapore. L.T. and Y.-Q.X. were supported by the ISREC Foundation with a donation from the Biltema Foundation and Swiss National Science Foundation (project grant 315230_173243). M.V.M. was supported by NIH grant CA K08166039. D.J.I. is an investigator of the Howard Hughes Medical Institute.

Author information

Author notes

    • Li Tang

    Present addresses: Institute of Bioengineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland; Institute of Materials Science and Engineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.

    • Li Tang
    •  & Yiran Zheng

    These authors contributed equally to this work.


  1. David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA.

    • Li Tang
    • , Yiran Zheng
    • , Mariane Bandeira Melo
    • , Llian Mabardi
    • , Na Li
    •  & Darrell J Irvine
  2. Department of Materials Science and Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Li Tang
    •  & Darrell J Irvine
  3. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Li Tang
    • , Yiran Zheng
    • , Mariane Bandeira Melo
    • , Na Li
    •  & Darrell J Irvine
  4. Cellular Immunotherapy Program, Massachusetts General Hospital (MGH) Cancer Center, Charlestown, Massachusetts, USA.

    • Ana P Castaño
    •  & Marcela V Maus
  5. Institute of Bioengineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.

    • Yu-Qing Xie
  6. Vaccine Center, Wistar Institute, Philadelphia, Pennsylvania, USA.

    • Sagar B Kudchodkar
  7. Altor BioScience Corporation, Miramar, Florida, USA.

    • Hing C Wong
    •  & Emily K Jeng
  8. Harvard Medical School, Boston, Massachusetts, USA.

    • Marcela V Maus
  9. Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Darrell J Irvine
  10. Howard Hughes Medical Institute, Chevy Chase, Maryland, USA.

    • Darrell J Irvine


  1. Search for Li Tang in:

  2. Search for Yiran Zheng in:

  3. Search for Mariane Bandeira Melo in:

  4. Search for Llian Mabardi in:

  5. Search for Ana P Castaño in:

  6. Search for Yu-Qing Xie in:

  7. Search for Na Li in:

  8. Search for Sagar B Kudchodkar in:

  9. Search for Hing C Wong in:

  10. Search for Emily K Jeng in:

  11. Search for Marcela V Maus in:

  12. Search for Darrell J Irvine in:


L.T., Y.Z., M.B.M. and D.J.I. designed the in vitro and syngeneic mouse experiments; H.C.W. and E.K.J. provided ALT-803; L.T., Y.Z., D.J.I., A.P.C., S.B.K. and M.V.M. designed the studies with the humanized mice; L.T., Y.Z., L.M., M.B.M., Y.-Q.X., N.L., A.P.C. and S.B.K. performed the experiments; L.T., Y.Z., M.B.M. and D.J.I. analyzed the data and wrote the manuscript; and all authors edited the manuscript.

Competing interests

D.J.I., L.T., and Y.Z. are inventors on licensed patents related to the technology described in this manuscript. D.J.I. is a co-founder of Torque Therapeutics, which licensed patents related to this technology.

Corresponding authors

Correspondence to Li Tang or Darrell J Irvine.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–17, Supplementary Tables 1–3, and Supplementary Scheme 1

  2. 2.

    Life Sciences Reporting Summary

About this article

Publication history