Cancer Immunology, Immunotherapy

, Volume 67, Issue 5, pp 761–774 | Cite as

TriCurin, a synergistic formulation of curcumin, resveratrol, and epicatechin gallate, repolarizes tumor-associated macrophages and triggers an immune response to cause suppression of HPV+ tumors

  • Sumit Mukherjee
  • Rahman Hussaini
  • Richard White
  • Doaa Atwi
  • Angela Fried
  • Samay Sampat
  • Longzhu Piao
  • Quintin Pan
  • Probal Banerjee
Original Article


Our earlier studies reported a unique potentiated combination (TriCurin) of curcumin (C) with two other polyphenols. The TriCurin-associated C displays an IC50 in the low micromolar range for cultured HPV+ TC-1 cells. In contrast, because of rapid degradation in vivo, the TriCurin-associated C reaches only low nano-molar concentrations in the plasma, which are sub-lethal to tumor cells. Yet, injected TriCurin causes a dramatic suppression of tumors in TC-1 cell-implanted mice (TC-1 mice) and xenografts of Head and Neck Squamous Cell Carcinoma (HNSCC) cells in nude/nude mice. Here, we use the TC-1 mice to test our hypothesis that a major part of the anti-tumor activity of TriCurin is evoked by innate and adaptive immune responses. TriCurin injection repolarized arginase1high (ARG1high), IL10high, inducible nitric oxide synthaselow (iNOSlow), IL12low M2-type tumor-associated macrophages (TAM) into ARG1low, IL10low, iNOShigh, and IL12high M1-type TAM in HPV+ tumors. The M1 TAM displayed sharply suppressed STAT3 and induced STAT1 and NF-kB(p65). STAT1 and NF-kB(p65) function synergistically to induce iNOS and IL12 transcription. Neutralizing IL12 signaling with an IL12 antibody abrogated TriCurin-induced intra-tumor entry of activated natural killer (NK) cells and Cytotoxic T lymphocytes (CTL), thereby confirming that IL12 triggers recruitment of NK cells and CTL. These activated NK cells and CTL join the M1 TAM to elicit apoptosis of the E6+ tumor cells. Corroboratively, neutralizing IL12 signaling partially reversed this TriCurin-mediated apoptosis. Thus, injected TriCurin elicits an M2→M1 switch in TAM, accompanied by IL12-dependent intra-tumor recruitment of NK cells and CTL and elimination of cancer cells.


Curcumin TriCurin Papillomavirus Tumor-associated macrophages NK cells CTL 







Cluster of differentiation 8 protein


Cluster of differentiation 8 alpha


Cytotoxic T lymphocytes


City University of New York


Epicatechin gallate

E6 16/18

Early 6 protein 16/18


Early 7 protein


Harvey ras protein


Hydrochloric acid


Head and neck squamous cell carcinoma


Human papillomavirus


Institutional Animal Care and Use Committee


Ionized calcium-binding adapter molecule 1


Integrated fluorescence


Interferon gamma


Immunoglobin G






Inducible nitric oxide synthase


Loop electrosurgical excision procedure


Lower right


Athymic nude mice


Natural killer cell p-46-related protein


Nitric oxide synthase 2
















Standard error


Standard error of the mean


Saline-sodium citrate


Tumor-associated macrophage

TriCur + IL12Ab

‘IL12 antibody infusion followed by TriCurin treatment’ group


Upper left


University of Michigan-squamous cell carcinoma-47


Upper right





Sumit Mukherjee was supported by a teaching assistantship from the College of Staten Island (CUNY).

Author contributions

PB, SM, QP, and LP conceived and planned the experiments. SM, RH, RW, DA, AF, SS, and LP carried out the experiments. All the authors contributed to the interpretation of results. SM and PB took the lead in writing the manuscript. All authors provided critical feedback and helped in shaping the final manuscript.


This project was supported by Professional Staff Congress (PSC)-CUNY Grants from cycles 45 and 47.

Compliance with ethical standards

Conflict of interest

Probal Banerjee has a pending patent application “Activity Enhancing Curcumin Compositions and Methods of Use”, PCT/US14/67819 pending. The remaining authors declare no conflict of interest.

Ethical approval and ethical standards

All procedures performed in studies involving animals were in accordance with the ethical standards of the Institutional Animal Care Committees (IACUC) at the College of Staten Island (approval # 11 − 008) and The Ohio State University Medical Center (approval # 2009A0172).

Animal source

Adult C57BL/6 female mice (2–6 months old) used for the TC-1 tumor experiments were bred and handled at the College of Staten Island (CUNY) [8]. Athymic nude (NCr) mice were purchased from Charles River Laboratories (Wilmington, MA) for the Head and Neck Squamous Cell Carcinoma (HNSCC) UMSCC47 tumor xenograft model and housed at The Ohio State University animal facility [7]. Both the strains mentioned above were bred and handled in accordance to the consent of respective Institutional Animal Care Committees (IACUC) at the College of Staten Island and The Ohio State University Medical Center. All animal protocols were approved by the respective Institutional Animal Care Committees (IACUC) at the College of Staten Island and The Ohio State University Medical Center.

Supplementary material

262_2018_2130_MOESM1_ESM.pdf (1.9 mb)
Supplementary material 1 (PDF 1973 KB)


  1. 1.
    Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D (2011) Global cancer statistics CA. Cancer J Clin 61:69–90CrossRefGoogle Scholar
  2. 2.
    Kamangar F, Dores GM, Anderson WF (2006) Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol 24:2137–2150. CrossRefPubMedGoogle Scholar
  3. 3.
    Fowler RS (2000) Vulvar vestibulitis: response to hypocontactant vulvar therapy. J Low Genit Tract Dis 4:200–203CrossRefPubMedGoogle Scholar
  4. 4.
    Stanley M (2010) Potential mechanisms for HPV vaccine-induced long-term protection. Gynecol Oncol 118:2–7. CrossRefGoogle Scholar
  5. 5.
    Leemans CR, Braakhuis BJM, Brakenhoff RH (2011) The molecular biology of head and neck cancer. Nat Rev Cancer 11:9–22. CrossRefPubMedGoogle Scholar
  6. 6.
    Mirghani H, Amen F, Blanchard P, Moreau F, Guigay J, Hartl DM, Lacau St Guily J (2015) Treatment de-escalation in HPV-positive oropharyngeal carcinoma: ongoing trials, critical issues and perspectives. Int J Cancer 136:1494–1503. CrossRefPubMedGoogle Scholar
  7. 7.
    Piao L, Mukherjee S, Chang Q et al (2017) TriCurin, a novel formulation of curcumin, epicatechin gallate, and resveratrol, inhibits the tumorigenicity of human papillomavirus-positive head and neck squamous cell carcinoma. Oncotarget 8:60025–60035. PubMedGoogle Scholar
  8. 8.
    Mukherjee S, Debata PR, Hussaini R et al (2017) Unique synergistic formulation of curcumin, epicatechin gallate and resveratrol, tricurin, suppresses HPV E6, eliminates HPV+ cancer cells, and inhibits tumor progression. Oncotarget 8:60904–60916. PubMedPubMedCentralGoogle Scholar
  9. 9.
    Anand P, Kunnumakkara AB, Newman RA, Aggarwal BB (2007) Bioavailability of curcumin: problems and promises. Mol Pharm 4:807–818. CrossRefPubMedGoogle Scholar
  10. 10.
    Langone P, Debata PR, Dolai S, Curcio GM, Inigo JD, Raja K, Banerjee P (2012) Coupling to a cancer cell-specific antibody potentiates tumoricidal properties of curcumin. Int J Cancer 131:E569–E578. CrossRefPubMedGoogle Scholar
  11. 11.
    Langone P, Debata PR, Inigo JDR, Dolai S, Mukherjee S, Halat P, Mastroianni K, Curcio GM, Castellanos MR, Raja K, Banerjee P (2014) Coupling to a glioblastoma-directed antibody potentiates anti-tumor activity of curcumin. Int J Cancer 135:710–719CrossRefPubMedGoogle Scholar
  12. 12.
    Mukherjee S, Baidoo J, Fried A, Atwi D, Dolai S, Boockvar J, Symons M, Ruggieri R, Raja K, Banerjee P (2016) Curcumin changes the polarity of tumor-associated microglia and eliminates glioblastoma. Int J Cancer 139:2838–2849. CrossRefPubMedGoogle Scholar
  13. 13.
    Zhang X, Tian W, Cai X, Wang X, Dang X, Dang W, Tang H, Cao H, Wang L, Chen T (2013) Hydrazinocurcumin encapsuled nanoparticles “Re-Educate” tumor-associated macrophages and exhibit anti-tumor effects on breast cancer following STAT3 suppression. PLoS One 8:e65896CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Debata PR, Castellanos MR, Fata JE, Baggett S, Rajupet S, Szerszen A, Begum S, Mata A, Murty VV, Opitz LM, Banerjee P (2013) A novel curcumin-based vaginal cream Vacurin selectively eliminates apposed himan cervical cancer cells. Gynecol Oncol. 129:145–153CrossRefPubMedGoogle Scholar
  15. 15.
    Purkayastha S, Berliner A, Fernando SS, Ranasinghe B, Ray I, Tariq H, Banerjee P (2009) Curcumin blocks brain tumor formation. Brain Res 1266:130–138. CrossRefPubMedGoogle Scholar
  16. 16.
    Zhang HG, Kim H, Liu C, Yu S, Wang J, Grizzle WE, Kimberly RP, Barnes S (2007) Curcumin reverses breast tumor exosomes mediated immune suppression of NK cell tumor cytotoxicity. Biochim Biophys Acta 1773:1116–1123. CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Bhattacharyya S, Md Sakib Hossain D, Mohanty S et al. (2010) Curcumin reverses T cell-mediated adaptive immune dysfunctions in tumor-bearing hosts. Cell Mol Immunol 7:306–315. CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Chang YF, Chuang HY, Hsu CH, Liu RS, Gambhir SS, Hwang JJ (2012) Immunomodulation of curcumin on adoptive therapy with T cell functional imaging in mice. Cancer Prev Res (Phila.) 5:444–452. CrossRefGoogle Scholar
  19. 19.
    Luo F, Song X, Zhang Y, Chu Y (2011) Low-dose curcumin leads to the inhibition of tumor growth via enhancing CTL-mediated antitumor immunity. Int Immunopharmacol 11:1234–1240. CrossRefPubMedGoogle Scholar
  20. 20.
    Lu Y, Miao L, Wang Y, Xu Z, Zhao Y, Shen Y, Xiang G, Huang L (2016) Curcumin micelles remodel tumor microenvironment and enhance vaccine activity in an advanced melanoma model. Mol Ther 24:364–374. CrossRefPubMedGoogle Scholar
  21. 21.
    Farag SS, Caligiuri MA (2006) Human natural killer cell development and biology. Blood Rev 20:123–137. CrossRefPubMedGoogle Scholar
  22. 22.
    Postow MA, Callahan MK, Wolchok JD (2015) Immune checkpoint blockade in cancer therapy. J Clin Oncol 33:1974–1982. CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Zhang H, Ye Z-l, Yuan Z-G, Luo Z-Q, Jin H-J, Qian Q-J (2016) New strategies for the treatment of solid tumors with CAR-T Cells. Int J Biol Sci 12:718–729. CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Scharton-Kersten T, Afonso L, Wysocka M, Trinchieri G, Scott P (1995) IL-12 is required for natural killer cell activation and subsequent T helper 1 cell development in experimental leishmaniasis. J Immunol 154:5320–5330PubMedGoogle Scholar
  25. 25.
    Huang Z, Peng S, Knoff J, Lee SY, Yang B, Wu T-C, Hung C-F (2015) Combination of proteasome and HDAC inhibitor enhances HPV16 E7-specific CD8+ T cell immune response and antitumor effects in a preclinical cervical cancer model. J Biomed Sci 22:7CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Brantley EC, Guo L, Zhang C, Lin Q, Yokoi K, Langley RR, Kruzel E, Maya M, Kim SW, Kim S-J, Fan D, Fidler IJ (2010) Nitric oxide-mediated tumoricidal activity of murine microglial cells. Transl Oncol 3:380–388CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Chakravarti N, Myers JN, Aggarwal BB (2006) Targeting constitutive and interleukin-6-inducible signal transducers and activators of transcription 3 pathway in head and neck squamous cell carcinoma cells by curcumin (diferuloylmethane). Int J Cancer 119:1268–1275CrossRefPubMedGoogle Scholar
  28. 28.
    Ito S, Ansari P, Sakatsume M, Dickensheets H, Vazquez N, Donnelly RP, Larner AC, Finbloom DS (1999) Interleukin-10 inhibits expression of both interferon a– and interferon gamma–induced genes by suppressing tyrosine phosphorylation of STAT1. Blood 93:1456–1463PubMedGoogle Scholar
  29. 29.
    Hagemann T, Lawrence T, McNeish I, Charles KA, Kulbe H, Thompson RG, Robinson SC, Balkwill FR (2008) “Re-educating” tumor-associated macrophages by targeting NF-kB. J Exp Med 205:1261–1268CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Hagemann T, Biswas SK, Lawrence T, Sica A, Lewis CE (2009) Regulation of macrophage function in tumors: the multifaceted role of NF-kappaB. Blood 113:3139–3146. CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Ohmori Y, Hamilton TA (2001) Requirement for STAT1 in LPS-induced geneexpression in macrophages. J Leukoc Biol 69:598–604PubMedGoogle Scholar
  32. 32.
    Vakkila J, Demarco RA, Lotze MT (2008) Coordinate NF-kB and STAT1 activation promotes development of myeloid type 1 dendritic cells. Scand J Immunol 67:260–269CrossRefPubMedGoogle Scholar
  33. 33.
    Marotta LLC, Almendro V, Marusyk A et al (2011) The JAK2/STAT3 signaling pathway is required for growth of CD44+ CD24– stem cell–like breast cancer cells in human tumors. J Clin Investig 121:2723–2735. CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Schroder K, Hertzog PJ, Ravasi T, Hume DA (2004) Interferon-gamma: an overview of signals, mechanisms and functions. J Leukoc Biol 75:163–189. CrossRefPubMedGoogle Scholar
  35. 35.
    Bontkes HJ, Kramer D, Ruizendaal JJ, Meijer CJ, Hooijberg E (2008) Tumor associated antigen and interleukin-12 mRNA transfected dendritic cells enhance effector function of natural killer cells and antigen specific T-cells. Clin Immunol 127:375–384CrossRefPubMedGoogle Scholar
  36. 36.
    Hamza T, Barnett JB, Li B (2010) Interleukin 12 a key immunoregulatory cytokine in infection applications. Int J Mol Sci 11:789–806. CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Kuge S, Watanabe K, Makino K, Tokuda Y, Mitomi T, Kawamura N, Habu S, Nishimura T (1995) Interleukin-12 augments the generation of autologous tumor-reactive CD8+ cytotoxic T lymphocytes from tumor-infiltrating lymphocytes. Jpn J Cancer Res 86:135–139CrossRefPubMedGoogle Scholar
  38. 38.
    Michel T, Hentges F, Zimmer J (2012) Consequences of the crosstalk between monocytes/macrophages and natural killer cells. Front Immunol 3:403. PubMedGoogle Scholar
  39. 39.
    Morrison BE, Park SJ, Mooney JM, Mehrad B (2003) Chemokine-mediated recruitment of NK cells is a critical host defense mechanism in invasive aspergillosis. J Clin Invest 112:1862–1870. CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Gertsch J, Guttinger M, Heilmann J, Sticher O (2003) Curcumin differentially modulates mRNA profiles in Jurkat T and human peripheral blood mononuclear cells. Bioorg Med Chem 11:1057–1063CrossRefPubMedGoogle Scholar
  41. 41.
    Kang TH, Lee JH, Song CK et al (2007) Epigallocatechin-3-gallate enhances CD8+ T cell-mediated antitumor immunity induced by DNA vaccination. Cancer Res 67:802–811. CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Bellora F, Castriconi R, Dondero A, Reggiardo G, Moretta L, Mantovani A, Moretta A, Bottino C (2010) The interaction of human natural killer cells with either unpolarized or polarized macrophages results in different functional outcomes. Proc Natl Acad Sci USA 107:21659–21664CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Verma R, Foster RE, Horgan K, Mounsey K, Nixon H, Smalle N, Hughes TA, Carter CR (2016) Lymphocyte depletion and repopulation after chemotherapy for primary breast cancer. Breast Cancer Res 18:10. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.CUNY Doctoral Program in BiochemistryCUNY Graduate CenterNew YorkUSA
  2. 2.Department of Chemistry, Building 6SThe City University of New York at The College of Staten IslandStaten IslandUSA
  3. 3.The Center for Developmental NeuroscienceCity University of New York at The College of Staten IslandStaten IslandUSA
  4. 4.College of Arts and ScienceNew York UniversityNew YorkUSA
  5. 5.Department of Otolaryngology-Head and Neck SurgeryThe Ohio State University Medical CenterColumbusUSA
  6. 6.Arthur G. James Cancer Hospital and Richard J. Solove Research InstituteThe Ohio State University Comprehensive Cancer CenterColumbusUSA

Personalised recommendations