IL-6 trans-signaling promotes the expansion and anti-tumor activity of CAR T cells


Chimeric antigen receptor (CAR) T cell therapies lead to high clinical response rates in B cell malignancies, and are under investigation for treatment of solid tumors. While high systemic interleukin- (IL-) 6 levels are associated with clinical cytokine release syndrome (CRS), the role of IL-6 trans-signaling within CAR T-cells has not been reported. We generated CAR T cells that constitutively express hyper IL-6 (HIL-6), a designer cytokine that activates the trans-signaling pathway. HIL-6-expressing CAR T-cells exhibited enhanced proliferation and antitumor efficacy in vitro and in xenograft models. However, HIL-6 CAR T cells caused severe graft-versus-host disease (GVHD). Transcriptomic profiling revealed that HIL-6 stimulation of CAR T cells upregulated genes associated with T cell migration, early memory differentiation, and IL-6/GP130/STAT3 signaling. Since IL-6 trans-signaling acts via surface GP130, we generated CAR T cells expressing a constitutively-active form of GP130 and found these retained improved antitumor activity without signs of GVHD in preclinical models of B-cell leukemia and solid tumors. Taken together, these results show that IL-6 trans-signaling can enhance expansion and antitumor activity of CAR T cells via the GP130/STAT3 pathway, and suggest that expression of GP130 within CAR T cells could lead to improved antitumor efficacy without systemic IL-6 trans-signaling.

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Fig. 1: HIL-6 improves the expansion, survival and cytotoxicity of CAR T cells after stimulation by target cells in vitro.
Fig. 2: CAR19/HIL-6 suppressed growth of leukemia but led to development of GVHD in a dose-dependent fashion.
Fig. 3: Cytotoxicity of HIL-6 CAR T cells against lung cancer and hepatocellular carcinoma in xenograft models.
Fig. 4: CAR T cell treatment with HIL-6 upregulates expression of genes associated with cell migration, memory differentiation, and the IL-6/GP130/STAT3 pathway.
Fig. 5: GP130 signaling leads to improve CAR T cell antitumor efficacy.


  1. 1.

    Locke FL, Ghobadi A, Jacobson CA, Miklos DB, Lekakis LJ, Oluwole OO, et al. Long-term safety and activity of axicabtagene ciloleucel in refractory large B-cell lymphoma (ZUMA-1): a single-arm, multicentre, phase 1-2 trial. Lancet Oncol. 2019;20:31–42.

    CAS  Article  PubMed  Google Scholar 

  2. 2.

    Maude SL, Laetsch TW, Buechner J, Rives S, Boyer M, Bittencourt H, et al. Tisagenlecleucel in Children and Young Adults with B-Cell Lymphoblastic Leukemia. N Engl J Med. 2018;378:439–48.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  3. 3.

    Brown CE, Alizadeh D, Starr R, Weng L, Wagner JR, Naranjo A, et al. Regression of glioblastoma after chimeric antigen receptor T-cell therapy. N Engl J Med. 2016;375:2561–9.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    Li J, Li W, Huang K, Zhang Y, Kupfer G, Zhao Q. Chimeric antigen receptor T cell (CAR-T) immunotherapy for solid tumors: lessons learned and strategies for moving forward. J Hematol Oncol. 2018;11:22.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  5. 5.

    Kagoya Y, Tanaka S, Guo T, Anczurowski M, Wang CH, Saso K, et al. A novel chimeric antigen receptor containing a JAK-STAT signaling domain mediates superior antitumor effects. Nat Med. 2018;24:352–9.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Lai Y, Weng J, Wei X, Qin L, Lai P, Zhao R, et al. Toll-like receptor 2 costimulation potentiates the antitumor efficacy of CAR T Cells. Leukemia. 2018;32:801–8.

    CAS  Article  PubMed  Google Scholar 

  7. 7.

    Koneru M, O’Cearbhaill R, Pendharkar S, Spriggs DR, Brentjens RJ. A phase I clinical trial of adoptive T cell therapy using IL-12 secreting MUC-16(ecto) directed chimeric antigen receptors for recurrent ovarian cancer. J Transl Med. 2015;13:102.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. 8.

    Shum T, Omer B, Tashiro H, Kruse RL, Wagner DL, Parikh K, et al. Constitutive signaling from an engineered IL7 receptor promotes durable tumor elimination by tumor-redirected T cells. Cancer Disco. 2017;7:1238–47.

    CAS  Article  Google Scholar 

  9. 9.

    Krenciute G, Prinzing BL, Yi Z, Wu MF, Liu H, Dotti G, et al. Transgenic expression of IL15 improves antiglioma activity of IL13Ralpha2-CAR T cells but results in antigen loss variants. Cancer Immunol Res. 2017;5:571–81.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  10. 10.

    Teachey DT, Lacey SF, Shaw PA, Melenhorst JJ, Maude SL, Frey N, et al. Identification of predictive biomarkers for cytokine release syndrome after chimeric antigen receptor T-cell therapy for acute lymphoblastic leukemia. Cancer Disco. 2016;6:664–79.

    CAS  Article  Google Scholar 

  11. 11.

    Porter D, Frey N, Wood PA, Weng Y, Grupp SA. Grading of cytokine release syndrome associated with the CAR T cell therapy tisagenlecleucel. J Hematol Oncol. 2018;11:35.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. 12.

    Fraietta JA, Lacey SF, Orlando EJ, Pruteanu-Malinici I, Gohil M, Lundh S, et al. Determinants of response and resistance to CD19 chimeric antigen receptor (CAR) T cell therapy of chronic lymphocytic leukemia. Nat Med. 2018;24:563–71.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. 13.

    Wang L, Miyahira AK, Simons DL, Lu X, Chang AY, Wang C, et al. IL6 signaling in peripheral blood T cells predicts clinical outcome in breast cancer. Cancer Res. 2017;77:1119–26.

    CAS  Article  Google Scholar 

  14. 14.

    Lee DW, Gardner R, Porter DL, Louis CU, Ahmed N, Jensen M, et al. Current concepts in the diagnosis and management of cytokine release syndrome. Blood. 2014;124:188–95.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  15. 15.

    Mueller KT, Waldron E, Grupp SA, Levine JE, Laetsch TW, Pulsipher MA, et al. Clinical pharmacology of tisagenlecleucel in B-cell acute lymphoblastic leukemia. Clin Cancer Res. 2018;24:6175–84.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. 16.

    Stein AM, Grupp SA, Levine JE, Laetsch TW, Pulsipher MA, Boyer MW, et al. Tisagenlecleucel model-based cellular kinetic analysis of chimeric antigen receptor-T cells. CPT Pharmacomet Syst Pharm. 2019;8:285–95.

    CAS  Article  Google Scholar 

  17. 17.

    Tanaka T, Narazaki M, Kishimoto T. IL-6 in inflammation, immunity, and disease. Cold Spring Harb Perspect Biol. 2014;6:a016295.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. 18.

    Jones SA, Scheller J, Rose-John S. Therapeutic strategies for the clinical blockade of IL-6/gp130 signaling. J Clin Invest. 2011;121:3375–83.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Bottcher JP, Schanz O, Garbers C, Zaremba A, Hegenbarth S, Kurts C, et al. IL-6 trans-signaling-dependent rapid development of cytotoxic CD8+ T cell function. Cell Rep. 2014;8:1318–27.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. 20.

    McLoughlin RM, Jenkins BJ, Grail D, Williams AS, Fielding CA, Parker CR, et al. IL-6 trans-signaling via STAT3 directs T cell infiltration in acute inflammation. Proc Natl Acad Sci USA. 2005;102:9589–94.

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Atreya R, Mudter J, Finotto S, Mullberg J, Jostock T, Wirtz S, et al. Blockade of interleukin 6 trans signaling suppresses T-cell resistance against apoptosis in chronic intestinal inflammation: evidence in crohn disease and experimental colitis in vivo. Nat Med. 2000;6:583–8.

    CAS  Article  PubMed  Google Scholar 

  22. 22.

    Singh N, Hofmann TJ, Gershenson Z, Levine BL, Grupp SA, Teachey DT, et al. Monocyte lineage-derived IL-6 does not affect chimeric antigen receptor T-cell function. Cytotherapy. 2017;19:867–80.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  23. 23.

    Weng J, Lai P, Qin L, Lai Y, Jiang Z, Luo C, et al. A novel generation 1928zT2 CAR T cells induce remission in extramedullary relapse of acute lymphoblastic leukemia. J Hematol Oncol. 2018;11:25.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. 24.

    Wei X, Lai Y, Li J, Qin L, Xu Y, Zhao R, et al. PSCA and MUC1 in non-small-cell lung cancer as targets of chimeric antigen receptor T cells. Oncoimmunology. 2017;6:e1284722.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Jiang Z, Jiang X, Chen S, Lai Y, Wei X, Li B, et al. Anti-GPC3-CAR T cells suppress the growth of tumor cells in patient-derived xenografts of hepatocellular carcinoma. Front Immunol. 2016;7:690.

    Article  CAS  PubMed  Google Scholar 

  26. 26.

    Fischer M, Goldschmitt J, Peschel C, Brakenhoff JP, Kallen KJ, Wollmer A, et al. I. A bioactive designer cytokine for human hematopoietic progenitor cell expansion. Nat Biotechnol. 1997;15:142–5.

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Stuhlmann-Laeisz C, Lang S, Chalaris A, Krzysztof P, Enge S, Eichler J, et al. Forced dimerization of gp130 leads to constitutive STAT3 activation, cytokine-independent growth, and blockade of differentiation of embryonic stem cells. Mol Biol Cell. 2006;17:2986–95.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  28. 28.

    Brown CE, Aguilar B, Starr R, Yang X, Chang WC, Weng L, et al. Optimization of IL13Ralpha2-targeted chimeric antigen receptor T cells for improved anti-tumor efficacy against glioblastoma. Mol Ther. 2018;26:31–44.

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Fisher DT, Chen Q, Skitzki JJ, Muhitch JB, Zhou L, Appenheimer MM, et al. IL-6 trans-signaling licenses mouse and human tumor microvascular gateways for trafficking of cytotoxic T cells. J Clin Invest. 2011;121:3846–59.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  30. 30.

    Ye W, Jiang Z, Li GX, Xiao Y, Lin S, Lai Y, et al. Quantitative evaluation of the immunodeficiency of a mouse strain by tumor engraftments. J Hematol Oncol. 2015;8:59.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. 31.

    Gattinoni L, Klebanoff CA, Restifo NP. Paths to stemness: building the ultimate antitumour T cell. Nat Rev Cancer. 2012;12:671–84.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  32. 32.

    Heinrich PC, Behrmann I, Muller-Newen G, Schaper F, Graeve L. Interleukin-6-type cytokine signalling through the gp130/Jak/STAT pathway. Biochem J. 1998;334:297–314.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  33. 33.

    Rivron NC, Frias-Aldeguer J, Vrij EJ, Boisset JC, Korving J, Vivie J, et al. Blastocyst-like structures generated solely from stem cells. Nature. 2018;557:106–11.

    CAS  Article  PubMed  Google Scholar 

  34. 34.

    O’Shea EK, Rutkowski R, Stafford WF 3rd, Kim PS. Preferential heterodimer formation by isolated leucine zippers from fos and jun. Science. 1989;245:646–8.

    Article  PubMed  Google Scholar 

  35. 35.

    Cheadle EJ, Hawkins RE, Batha H, O’Neill AL, Dovedi SJ, Gilham DE. Natural expression of the CD19 antigen impacts the long-term engraftment but not antitumor activity of CD19-specific engineered T cells. J Immunol. 2010;184:1885–96.

    CAS  Article  Google Scholar 

  36. 36.

    van der Stegen SJ, Davies DM, Wilkie S, Foster J, Sosabowski JK, Burnet J, et al. Preclinical in vivo modeling of cytokine release syndrome induced by ErbB-retargeted human T cells: identifying a window of therapeutic opportunity? J Immunol. 2013;191:4589–98.

    Article  CAS  Google Scholar 

  37. 37.

    Giavridis T, van der Stegen SJC, Eyquem J, Hamieh M, Piersigilli A, Sadelain MCAR. T cell-induced cytokine release syndrome is mediated by macrophages and abated by IL-1 blockade. Nat Med. 2018;24:731–8.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  38. 38.

    Mestermann K, Giavridis T, Weber J, Rydzek J, Frenz S, Nerreter T, et al. The tyrosine kinase inhibitor dasatinib acts as a pharmacologic on/off switch for CAR T cells. Sci Transl Med. 2019;11:eaau5907.

  39. 39.

    Norelli M, Camisa B, Barbiera G, Falcone L, Purevdorj A, Genua M, et al. Monocyte-derived IL-1 and IL-6 are differentially required for cytokine-release syndrome and neurotoxicity due to CAR T cells. Nat Med. 2018;24:739–48.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  40. 40.

    Davila ML, Riviere I, Wang X, Bartido S, Park J, Curran K, et al. Efficacy and toxicity management of 19-28z CAR T cell therapy in B cell acute lymphoblastic leukemia. Sci Transl Med. 2014;6:224ra225.

    Article  CAS  Google Scholar 

  41. 41.

    Ganetsky A, Frey NV, Hexner EO, Loren AW, Gill SI, Luger SM, et al. Tocilizumab for the treatment of severe steroid-refractory acute graft-versus-host disease of the lower gastrointestinal tract. Bone Marrow Transpl. 2019;54:212–7.

    CAS  Article  Google Scholar 

  42. 42.

    Tawara I, Koyama M, Liu C, Toubai T, Thomas D, Evers R, et al. Interleukin-6 modulates graft-versus-host responses after experimental allogeneic bone marrow transplantation. Clin Cancer Res. 2011;17:77–88.

    CAS  Article  PubMed  Google Scholar 

  43. 43.

    Jacoby E, Yang Y, Qin H, Chien CD, Kochenderfer JN, Fry TJ. Murine allogeneic CD19 CAR T cells harbor potent antileukemic activity but have the potential to mediate lethal GVHD. Blood. 2016;127:1361–70.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  44. 44.

    Wang XJ, Taga T, Yoshida K, Saito M, Kishimoto T, Kikutani H. gp130, the cytokine common signal-transducer of interleukin-6 cytokine family, is downregulated in T cells in vivo by interleukin-6. Blood. 1998;91:3308–14.

    CAS  Article  PubMed  Google Scholar 

  45. 45.

    Cho DC, Brennan HJ, Johnson RW, Poulton IJ, Gooi JH, Tonkin BA, et al. Bone corticalization requires local SOCS3 activity and is promoted by androgen action via interleukin-6. Nat Commun. 2017;8:806.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. 46.

    Tan MSY, Sandanaraj E, Chong YK, Lim SW, Koh LWH, Ng WH, et al. A STAT3-based gene signature stratifies glioma patients for targeted therapy. Nat Commun. 2019;10:3601.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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This study is supported by the Strategic Priority Research Program of the Chinese Academy of Sciences, Grant No.XDB19030205, No.XDA1205030; National Natural Science Foundation of China (NSFC), No.81773301, 81700156, 8187012, 81873847; the Youth Innovation Promotion Association of the Chinese Academy of Sciences (2020351); Guangdong Special Support Program, NO.2017TX04R102; the Frontier and key technology innovation special grant from the Department of Science and Technology of Guangdong province, No. 2015B020227003; the Natural Science Fund of Guangdong Province-Doctoral Foundation, No. 2017A030310381; Guangdong Laboratory of Regenerative Medicine and Health-Guangzhou Frontier Exploration Project, No.2018GZR110105003; Science and Technology Planning Project of Guangdong Province, China (2017B030314056); Science and Technology Program of Guangzhou, China (202002020083); Guangzhou Medical University High-level University Construction Research Startup Fund, NO. B195002004013.

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Contribution: PL, ZJ, RL, JL, and YY conceived and designed research; ZJ, RL, JL, XH, SC, SL, YL, QW, SW, SL, PS, PL, and YJ performed experiments, collected, and analyzed data; DW, LQ, RZ designed and constructed the CAR; YL, XD, WW, SC, TZ, PL, DP, YY, RW, YY, and PL wrote the manuscript; and all authors revised and approved the manuscript.

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Correspondence to Yao Yao or Peng Li.

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Jiang, Z., Liao, R., Lv, J. et al. IL-6 trans-signaling promotes the expansion and anti-tumor activity of CAR T cells. Leukemia (2020).

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