Molecular Medicine

, Volume 20, Issue 1, pp 372–380 | Cite as

Decreased Langerhans Cell Responses to IL-36γ: Altered Innate Immunity in Patients with Recurrent Respiratory Papillomatosis

  • James DeVoti
  • Lynda Hatam
  • Alexandra Lucs
  • Ali Afzal
  • Allan Abramson
  • Bettie Steinberg
  • Vincent Bonagura
Research Article


Recurrent respiratory papillomatosis (RRP) is a rare, chronic disease caused by human papillomaviruses (HPVs) types 6 and 11 that is characterized by the polarization of adaptive immune responses that support persistent HPV infection. Respiratory papillomas express elevated mRNA levels of IL-36γ, a proinflammatory cytokine in comparison to autologous clinically normal laryngeal tissues; however there is no evidence of inflammation in these lesions. Consistent with this, respiratory papillomas do not contain TH1-like CD4+ T-cells or cytotoxic CD8+ T-cells, but instead contain a predominance of TH2-like and T regulatory cells (Tregs). In addition, papillomas also are infiltrated with immature Langerhans cells (iLCs). In this study, we show that papilloma cells express IL-36γ protein, and that human keratinocytes transduced with HPV11 have reduced IL-36γ secretion. We now provide the first evidence that peripheral blood-derived iLCs respond to IL-36γ by expressing inflammatory cytokines and chemokines. When stimulated with IL-36γ, iLCs from patients with RRP had lower expression levels of the TH2-like chemokine CCL-20 as compared with controls. Patients’ iLCs also had decreased steady state levels of CCL-1, which is a proinflammatory chemokine. Moreover, CCL-1 levels in iLCs inversely correlated with the severity of RRP. The combined decrease of TH1- and a TH2-like chemokines by iLCs from patients could have consequences in the priming of IFN-γ expression by CD8+ T-cells. Taken together, our results suggest that, in RRP, there is a defect in the proinflammatory innate immune responses made by iLCs in response to IL-36γ. The consequence of this defect may lead to persistent HPV infection by failing to support an effective HPV-specific, TH1-like and/or Tc1-like adaptive response, thus resulting in the predominant TH2-like and/or Treg micromilieu present in papillomas.



The authors would like to acknowledge the helpful discussion and encouragement of Isaac Rodriguez-Chavez. Research reported in this publication was supported by the National Institute of Dental and Craniofacial Research of the National Institutes of Health under Award Number R01DE017227, and the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under Award Number R21AI105987, and by the Feinstein Institute for Medical Research, North Shore-LIJ Health System. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.


  1. 1.
    DeVoti JA, et al. (2004) Failure of gamma interferon but not interleukin-10 expression in response to human papillomavirus type 11 E6 protein in respiratory papillomatosis. Clin. Diagn. Lab. Immunol. 11:538–47.PubMedPubMedCentralGoogle Scholar
  2. 2.
    Bonagura VR, et al. (2009) KIR3DS1, KIR2DS1, and KIR2DS5 protect against the development of severe recurrent respiratory papillomatosis (RRP) in HPV-6/11-Infected patients. J. Allergy Clin. Immunol. 123:S165.CrossRefGoogle Scholar
  3. 3.
    Bonagura VR, et al. (2010) Recurrent respiratory papillomatosis: a complex defect in immune responsiveness to human papillomavirus-6 and -11. APMIS. 118:455–70.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    DeVoti JA, et al. (2008) Immune dysregulation and tumor-associated gene changes in recurrent respiratory papillomatosis: A paired microarray analysis. Mol. Med. 14:608–17.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Vigne S, et al. (2012) IL-36 signaling amplifies Th1 responses by enhancing proliferation and Th1 polarization of naive CD4+ T cells. Blood. 120:3478–87.CrossRefPubMedGoogle Scholar
  6. 6.
    Mutamba S, Allison A, Mahida Y, Barrow P, Foster N. (2011) Expression of IL-1Rrp2 by human myelomonocytic cells is unique to DCs and facilitates DC maturation by IL-1F8 and IL-1F9. Eur. J. Immunol. 42:607–17.CrossRefGoogle Scholar
  7. 7.
    Blumberg H, et al. (2007) Opposing activities of two novel members of the IL-1 ligand family regulate skin inflammation. J. Exp. Med. 204:2603–14.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Johnston A, et al. (2011) IL-1F5, -F6, -F8, and -F9: a novel IL-1 family signaling system that is active in psoriasis and promotes keratinocyte antimicrobial peptide expression. J. Immunol. 186:2613–22.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Vigne S, et al. (2011) IL-36R ligands are potent regulators of dendritic and T cells. Blood. 118:5813–23.CrossRefPubMedGoogle Scholar
  10. 10.
    Marrakchi S, et al. (2011) Interleukin-36-receptor antagonist deficiency and generalized pustular psoriasis. N. Engl. J. Med. 365:620–8.CrossRefPubMedGoogle Scholar
  11. 11.
    Towne JE, Sims JE. (2012) IL-36 in psoriasis. Curr. Opin. Pharmacol. 12:486–90.CrossRefPubMedGoogle Scholar
  12. 12.
    Dinarello CA. (2013) Overview of the interleukin-1 family of ligands and receptors. Semin. Immunol. 25:389–93.CrossRefPubMedGoogle Scholar
  13. 13.
    Carrier Y, et al. (2011) Inter-regulation of Th17 cytokines and the IL-36 cytokines in vitro and in vivo: implications in psoriasis pathogenesis. J. Invest. Dermatol. 131:2428–37.CrossRefPubMedGoogle Scholar
  14. 14.
    Steinberg BM, Abramson AL, Meade RP. (1982) Culture of human laryngeal papilloma cells in vitro. Otolaryngol. Head Neck Surg. 90:728–35.CrossRefPubMedGoogle Scholar
  15. 15.
    Kashima HB, Leventhal B, Mounts P, Papilloma Study Group. (1985) Scoring system to assess severity and course in recurrent respiratory papillomatosis. In: Papillomavirus: molecular and clinical aspects: proceedings of the Burroughs-Wellcome-UCLA Symposium held in Steamboat Springs, Colorado, April 8–14, 1985. Howley PM, Broker TR (eds). Alan R. Liss, New York.Google Scholar
  16. 16.
    Abramson AL, et al. (1992) Clinical effects of photodynamic therapy on recurrent laryngeal papillomas. Arch. Otolaryngol. Head Neck Surg. 118:25–29.CrossRefPubMedGoogle Scholar
  17. 17.
    Wu R, Coniglio SJ, Chan A, Symons MH, Steinberg BM. (2007) Up-regulation of Rac1 by epidermal growth factor mediates COX-2 expression in recurrent respiratory papillomas. Mol. Med. 13:143–50.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Kis-Toth K, et al. (2013) Monocyte-derived dendritic cell subpopulations use different types of matrix metalloproteinases inhibited by GM6001. Immunobiology. 218:1361–9.CrossRefPubMedGoogle Scholar
  19. 19.
    Chung DJ, et al. (2013) Langerhans-type and monocyte-derived human dendritic cells have different susceptibilities to mRNA electroporation with distinct effects on maturation and activation: implications for immunogenicity in dendritic cell-based immunotherapy. J. Transl. Med. 11:166.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Lian LH, Milora KA, Manupipatpong KK, Jensen LE. (2012) The double-stranded RNA analogue polyinosinic-polycytidylic acid induces keratinocyte pyroptosis and release of IL-36gamma. J. Invest. Dermatol. 132:1346–53.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Hatam LJ, et al. (2008) CD4(+)Foxp3(+)CD127(+low) T-Regulatory cells are increased in HPV infected papillomas in patients with recurrent respiratory papillomatosis (RRP). J. Allergy Clin. Immunol. 121:S211.CrossRefGoogle Scholar
  22. 22.
    Hatam LJ, et al. (2012) Immune suppression in premalignant respiratory papillomas: enriched functional CD4+Foxp3+ regulatory T cells and PD-1/PD-L1/L2 expression. Clin. Cancer Res. 18:1925–35.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Geissmann F, et al. (2002) Accumulation of immature Langerhans cells in human lymph nodes draining chronically inflamed skin. J. Exp. Med. 196:417–30.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Towne JE, Garka KE, Renshaw BR, Virca GD, Sims JE. (2004) Interleukin (IL)-1F6, IL-1F8, and IL-1F9 signal through IL-1Rrp2 and IL-1RAcP to activate the pathway leading to NF-kappaB and MAPKs. J. Biol. Chem. 279:13677–88.CrossRefPubMedGoogle Scholar
  25. 25.
    Zhang C, Zhang X, Chen XH. (2014) Inhibition of the interleukin-6 signaling pathway: a strategy to induce immune tolerance. Clin. Rev. Allergy Immunol. 2014, Mar 20 [Epub ahead of print].Google Scholar
  26. 26.
    Weng Z, Patel AB, Vasiadi M, Therianou A, Theoharides TC. (2014) Luteolin inhibits human keratinocyte activation and decreases NF-κB induction that is increased in psoriatic skin. PLoS One. 9:e90739.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Junk DJ, Bryson BL, Jackson MW. (2014) HiJAK’d signaling; the STAT3 paradox in senescence and cancer progression. Cancers (Basel). 6:741–55.CrossRefGoogle Scholar
  28. 28.
    Guggino G, et al. (2014) Targeting IL-6 signalling in early rheumatoid arthritis is followed by Th1 and Th17 suppression and Th2 expansion. Clin. Exp. Rheumatol. 32:77–81.PubMedGoogle Scholar
  29. 29.
    Bonagura VR, Hatam L, DeVoti J, Zeng FF, Steinberg BM. (1999) Recurrent respiratory papillomatosis: Altered CD8(+) T-cell subsets and T(H)1/T(H)2 cytokine imbalance. Clin. Immunol. 93:302–11.CrossRefPubMedGoogle Scholar
  30. 30.
    Rosenthal DW, et al. (2006) Recurrent respiratory papillomatosis (RRP): Increased T(H)2-like chemokine expression. J. Allergy Clin. Immunol. 117:S104.CrossRefGoogle Scholar
  31. 31.
    Rosenthal DW, Devoti JA, Steinberg BM, Abramson AL, Bonagura VR. (2012) TH2-like chemokine patterns correlate with disease severity in patients with recurrent respiratory papillomatosis. Mol. Med. 18:1338–45.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Bonagura VR, et al. (2004) HLA alleles, IFN-gamma responses to HPV-11 E6, and disease severity in patients with recurrent respiratory papillomatosis. Human Immunol. 65:773–82.CrossRefGoogle Scholar
  33. 33.
    Ramadas RA, Ewart SL, Medoff BD, LeVine AM. (2011) Interleukin-1 family member 9 stimulates chemokine production and neutrophil influx in mouse lungs. Am. J. Respir. Cell Mol. Biol. 44:134–45.CrossRefPubMedGoogle Scholar
  34. 34.
    Chustz RT, et al. (2011) Regulation and function of the IL-1 family Cytokine IL-1F9 in human bronchial epithelial cells. Am. J. Respir. Cell Mol. Biol. 45:145–53.CrossRefPubMedGoogle Scholar
  35. 35.
    Towne JE, et al. (2011) Interleukin-36 (IL-36) ligands require processing for full agonist (IL-36alpha, IL-36beta, and IL-36gamma) or antagonist (IL-36Ra) activity. J. Biol. Chem. 286:42594–602.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Steinman RM. (1991) The dendritic cell system and its role in immunogenicity. Annu. Rev. Immunol. 9:271–96.CrossRefPubMedGoogle Scholar
  37. 37.
    Hart DN. (1997) Dendritic cells: unique leukocyte populations which control the primary immune response. Blood. 90:3245–87.PubMedGoogle Scholar
  38. 38.
    Matzinger P. (1994) Tolerance, danger, and the extended family. Annu. Rev. Immunol. 12:991–1045.CrossRefPubMedGoogle Scholar
  39. 39.
    Noel W, et al. (2004) Alternatively activated macrophages during parasite infections. Trends Parasitol. 20:126–33.CrossRefPubMedGoogle Scholar
  40. 40.
    McGuirk P, McCann C, Mills KH. (2002) Pathogen-specific T regulatory 1 cells induced in the respiratory tract by a bacterial molecule that stimulates interleukin 10 production by dendritic cells: a novel strategy for evasion of protective T helper type 1 responses by Bordetella pertussis. J. Exp. Med. 195:221–31.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Shortman K, Liu YJ. (2002) Mouse and human dendritic cell subtypes. Nat. Rev. Immunol. 2:151–61.CrossRefPubMedGoogle Scholar
  42. 42.
    Shirakata Y. (2010) Regulation of epidermal keratinocytes by growth factors. J. Dermatol. Sci. 59:73–80.CrossRefPubMedGoogle Scholar
  43. 43.
    Werner S, Krieg T, Smola H. (2007) Keratinocyte-fibroblast interactions in wound healing. J. Invest. Dermatol. 127:998–1008.CrossRefPubMedGoogle Scholar
  44. 44.
    Pastore S, Mascia F, Girolomoni G. (2006) The contribution of keratinocytes to the pathogenesis of atopic dermatitis. Eur. J. Dermatol. 16:125–31.PubMedGoogle Scholar
  45. 45.
    Bergmann C, et al. (2008) T regulatory type 1 cells in squamous cell carcinoma of the head and neck: mechanisms of suppression and expansion in advanced disease. Clin. Cancer Res. 14:3706–15.CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Banchereau J, et al. (2000) Immunobiology of dendritic cells. Annu. Rev. Immunol. 18:767–811.CrossRefGoogle Scholar
  47. 47.
    Banchereau J, et al. (2003) Dendritic cells: controllers of the immune system and a new promise for immunotherapy. Ann. N. Y. Acad. Sci. 987:180–7.CrossRefPubMedGoogle Scholar
  48. 48.
    Caux C, et al. (1997) CD34+ hematopoietic progenitors from human cord blood differentiate along two independent dendritic cell pathways in response to granulocyte-macrophage colony-stimulating factor plus tumor necrosis factor alpha: II. Functional analysis. Blood. 90:1458–70.PubMedGoogle Scholar
  49. 49.
    Steinman RM, Turley S, Mellman I, Inaba K. (2000) The induction of tolerance by dendritic cells that have captured apoptotic cells. J. Exp. Med. 191:411–6.CrossRefPubMedPubMedCentralGoogle Scholar
  50. 50.
    Iwasaki A, Kelsall BL. (1999) Freshly isolated Peyer’s patch, but not spleen, dendritic cells produce interleukin 10 and induce the differentiation of T helper type 2 cells. J. Exp. Med. 190:229–39.CrossRefPubMedPubMedCentralGoogle Scholar
  51. 51.
    Enk AH, Angeloni VL, Udey MC, Katz SI. (1993) Inhibition of Langerhans cell antigen-presenting function by IL-10. A role for IL-10 in induction of tolerance. J. Immunol. 151:2390–8.PubMedGoogle Scholar
  52. 52.
    Beissert S, Hosoi J, Grabbe S, Asahina A, Granstein RD. (1995) IL-10 inhibits tumor antigen presentation by epidermal antigen-presenting cells. J. Immunol. 154:1280–6.PubMedGoogle Scholar
  53. 53.
    Romani N, et al. (2006) Epidermal Langerhans cells—changing views on their function in vivo. Immunol. Lett. 106:119–25.CrossRefPubMedGoogle Scholar
  54. 54.
    Elias AN, Nanda VS, Barr RJ. (2003) CD1a expression in psoriatic skin following treatment with propylthiouracil, an antithyroid thioureylene. BMC Dermatol. 3:3.CrossRefPubMedPubMedCentralGoogle Scholar
  55. 55.
    Kaplan DH, Jenison MC, Saeland S, Shlomchik WD, Shlomchik MJ. (2005) Epidermal Langerhans cell-deficient mice develop enhanced contact hypersensitivity. Immunity. 23:611–20.CrossRefPubMedGoogle Scholar
  56. 56.
    Klechevsky E, et al. (2008) Functional specializations of human epidermal Langerhans cells and CD14+ dermal dendritic cells. Immunity. 29:497–510.CrossRefPubMedPubMedCentralGoogle Scholar
  57. 57.
    Bennett CL, et al. (2011) Langerhans cells regulate cutaneous injury by licensing CD8 effector cells recruited to the skin. Blood. 117:7063–9.CrossRefPubMedPubMedCentralGoogle Scholar
  58. 58.
    James E, et al. (2010) Human papillomavirus (HPV)-specific T-cells recognizing dominant E2/E6 epitopes elicit reduced IFN-γ in patients with recurrent respiratory papillomatosis (RRP). J. Allergy Clin. Immunol. 125:S.Google Scholar
  59. 59.
    James EA, et al. (2011) Papillomavirus-specific CD4(+) T cells exhibit reduced STAT-5 signaling and altered cytokine profiles in patients with recurrent respiratory papillomatosis. J. Immunol. 186:6633–40.CrossRefPubMedPubMedCentralGoogle Scholar
  60. 60.
    Chopin M, et al. (2013) Langerhans cells are generated by two distinct PU.1–dependent transcriptional networks. J. Exp. Med. 210:2967–80.CrossRefPubMedPubMedCentralGoogle Scholar
  61. 61.
    Chopin M, Nutt SL. (2014) Establishing and maintaining the Langerhans cell network. Semin. Cell. Dev. Biol. 2014, Feb 7. [Epub ahead of print].Google Scholar
  62. 62.
    Bonagura VR, et al. (2010) Activating killer cell immunoglobulin-like receptors 3DS1 and 2DS1 protect against developing the severe form of re current respiratory papillomatosis. Human Immunol. 71:212–9.CrossRefGoogle Scholar
  63. 63.
    Gombert M, et al. (2005) CCL1–CCR8 interactions: an axis mediating the recruitment of T cells and Langerhans-type dendritic cells to sites of atopic skin inflammation. J. Immunol. 174:5082–91.CrossRefPubMedGoogle Scholar
  64. 64.
    Henry E, et al. (2008) Dendritic cells genetically engineered to express IL-10 induce long-lasting antigen-specific tolerance in experimental asthma. J. Immunol. 181:7230–42.CrossRefPubMedGoogle Scholar
  65. 65.
    Dieu-Nosjean MC, et al. (2000) Macrophage inflammatory protein 3alpha is expressed at inflamed epithelial surfaces and is the most potent chemokine known in attracting Langerhans cell precursors. J. Exp. Med. 192:705–18.CrossRefPubMedPubMedCentralGoogle Scholar
  66. 66.
    Merad M, et al. (2002) Langerhans cells renew in the skin throughout life under steady-state conditions. Nat. Immunol. 3:1135–41.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© The Author(s) 2014

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (

Authors and Affiliations

  • James DeVoti
    • 1
    • 3
  • Lynda Hatam
    • 1
    • 3
  • Alexandra Lucs
    • 1
    • 4
  • Ali Afzal
    • 2
  • Allan Abramson
    • 1
    • 4
  • Bettie Steinberg
    • 2
    • 4
  • Vincent Bonagura
    • 1
    • 2
    • 3
  1. 1.Feinstein Institute for Medical ResearchManhassetUSA
  2. 2.Elmezzi Graduate School of Molecular MedicineManhassetUSA
  3. 3.Division of Allergy and Immunology, Department of PediatricsHofstra North Shore-LIJ School of MedicineGreat NeckUSA
  4. 4.Department of OtolaryngologyHofstra North Shore-LIJ School of MedicineGreat NeckUSA

Personalised recommendations