Increased migration of Langerhans cells in response to HPV16 E6 and E7 oncogene silencing: role of CCL20

  • Jean-Hubert Caberg
  • Pascale Hubert
  • Ludivine Herman
  • Michael Herfs
  • Patrick Roncarati
  • Jacques Boniver
  • Philippe Delvenne
Original Article

Abstract

Human papillomavirus (HPV) infection, particularly type 16, is causally associated with cancer of the uterine cervix. The persistence or progression of cervical lesions suggests that viral antigens are not adequately presented to the immune system. This hypothesis is reinforced by the observation that most squamous intraepithelial lesions (SILs) show quantitative and functional alterations of Langerhans cells (LC). The infiltration of immature LC in the squamous epithelium is mainly controlled by Macrophage Inflammatory Protein 3α/CCL20. After having shown that CCL20 production is altered in HPV-transformed keratinocytes (KC), the possible role of HPV16 E6 and E7 viral oncoproteins in the reduced CCL20 levels observed in SILs was investigated by silencing HPV16 E6 and E7 oncogenes by RNA interference (siRNA). This treatment not only increased CCL20 secretion but also resulted in the modulation of NF-κB p50, p52 and p65 precursor localization. Moreover, silencing of E6 and E7 oncogenes in HPV16-transformed KC induced a significantly higher migratory capacity of LC in a Boyden chamber assay and in an in vitro formed (pre)neoplastic epithelium reminiscent of high-grade SILs. Anti-CCL20 neutralizing antibody experiments showed that the increased migration of LC is due to the re-expression of CCL20 in E6 and E7 siRNA transfected KC. These data suggest that HPV16 E6/E7-induced down-regulation of CCL20 observed during the cervical carcinogenesis may contribute to a diminished capacity of the immune system to control HPV infection.

Keywords

Langerhans cells CCL20 Human papillomavirus E6 E7 Organotypic culture Cell migration 

References

  1. 1.
    Al-Saleh W, Delvenne P, Arrese JE et al (1995) Inverse modulation of intraepithelial Langerhans cells and stromal macrophage dendrocyte populations in human papillomavirus-associated squamous intraepithelial lesions of the cervix. Virchows Arch 427:41–48PubMedCrossRefGoogle Scholar
  2. 2.
    Al-Saleh W, Giannini SL, Jacobs N et al (1998) Correlation of T-helper secretory differentiation and types of antigen-presenting cells in squamous intraepithelial lesions of the uterine cervix. J Pathol 184:283–290PubMedCrossRefGoogle Scholar
  3. 3.
    Avvakumov N, Torchia JMymryk JS (2003) Interaction of the HPV E7 proteins with the pCAF acetyltransferase. Oncogene 22:3833–3841PubMedCrossRefGoogle Scholar
  4. 4.
    Bell DF, Chomarat PF, Broyles DF et al (1999) In breast carcinoma tissue, immature dendritic cells reside within the tumor, whereas mature dendritic cells are located in peritumoral areas. J Exp Med 190:1417–1426PubMedCrossRefGoogle Scholar
  5. 5.
    Bernat A, Avvakumov N, Mymryk JS et al (2003) Interaction between the HPV E7 oncoprotein and the transcriptional coactivator p300. Oncogene 22:7871–7881PubMedCrossRefGoogle Scholar
  6. 6.
    Charbonnier AS, Rot A, Kohrgruber N et al (1999) MIP-3 alpha is involved in the constitutive trafficking of epidermal Langerhans cells. J Invest Dermatol 113:431–431Google Scholar
  7. 7.
    Cook DN, Prosser DM, Forster R et al (2000) CCR6 mediates dendritic cell localization, lymphocyte homeostasis, and immune responses in mucosal tissue. Immunity 12:495–503PubMedCrossRefGoogle Scholar
  8. 8.
    Delvenne P, AlSaleh W, Gilles C et al (1995) Inhibition of growth of normal and human papillomavirus-transformed keratinocytes in monolayer and organotypic cultures by interferon-gamma and tumor-necrosis-factor-alpha. Am J Pathol 146:589–598PubMedGoogle Scholar
  9. 9.
    Dieu-Nosjean MC, Massacrier C, Homey B et al (2000) Macrophage inflammatory protein 3 alpha is expressed at inflamed epithelial surfaces and is the most potent chemokine known in attracting Langerhans cell precursors. J Exp Med 192:705–717PubMedCrossRefGoogle Scholar
  10. 10.
    Dieu-Nosjean MC, Vicari A, Lebecque S et al (1999) Regulation of dendritic cell trafficking: a process that involves the participation of selective chemokines. J Leukoc Biol 66:252–262PubMedGoogle Scholar
  11. 11.
    Ellerbrock TV, Chiasson MA, Bush TJ et al (2000) Incidence of cervical squamous intraepithelial lesions in HIV-infected women. JAMA 283:1031–1037PubMedCrossRefGoogle Scholar
  12. 12.
    Gerritsen ME, Williams AJ, Neish AS et al (1997) CREB-binding protein/p300 are transcriptional coactivators of p65. Proc Natl Acad Sci USA 94:2927–2932PubMedCrossRefGoogle Scholar
  13. 13.
    Giannini SL, Hubert P, Doyen J et al (2002) Influence of the mucosal epithelium microenvironment on Langerhans cells: implications for the development of squamous intraepithelial lesions of the cervix. Int J Cancer 97:654–659PubMedCrossRefGoogle Scholar
  14. 14.
    Greaves DR, Wang W, Dairaghi DJ et al (1997) CCR6, a CC chemokine receptor that interacts with macrophage inflammatory protein 3alpha and is highly expressed in human dendritic cells. J Exp Med 186:837–844PubMedCrossRefGoogle Scholar
  15. 15.
    Guess JC, McCance DJ (2005) Decreased migration of Langerhans precursor-like cells in response to human keratinocytes expressing human papillomavirus type 16 E6/E7 is related to reduced macrophage inflammatory protein-3alpha production. J Virol 79:14852–14862PubMedCrossRefGoogle Scholar
  16. 16.
    Havard L, Rahmouni S, Boniver J et al (2005) High levels of p105 (NFKB1) and p100 (NFKB2) proteins in HPV16-transformed keratinocytes: role of E6 and E7 oncoproteins. Virology 331:357–366PubMedCrossRefGoogle Scholar
  17. 17.
    Hayden MS, Ghosh S (2004) Signaling to NF-kappaB. Genes Dev 18:2195–2224PubMedCrossRefGoogle Scholar
  18. 18.
    Herman L, Hubert P, Caberg JH et al (2006) MIP3 alpha stimulates the migration of Langerhans cells in models of human papillomavirus (HPV)-associated (pre) neoplastic epithelium. Cancer Immunol Immunother 56:1087–1096PubMedCrossRefGoogle Scholar
  19. 19.
    Homey B, Dieu-Nosjean MC, Wiesenborn A et al (2000) Up-regulation of macrophage inflammatory protein-3 alpha/CCL20 and CC chemokine receptor 6 in psoriasis. J Immunol 164:6621–6632PubMedGoogle Scholar
  20. 20.
    Huang SM, McCance DJ (2002) Down regulation of the interleukin-8 promoter by human papillomavirus type 16 E6 and E7 through effects on CREB binding protein/p300 and P/CAF. J Virol 76:8710–8721PubMedCrossRefGoogle Scholar
  21. 21.
    Hubert P, Bousarghin L, Greimers R et al (2005) Production of large numbers of Langerhans’ cells with intraepithelial migration ability in vitro. Exp Dermatol 14:469–477PubMedCrossRefGoogle Scholar
  22. 22.
    Hubert P, Caberg JH, Gilles C et al (2005) E-cadherin-dependent adhesion of dendritic and Langerhans cells to keratinocytes is defective in cervical human papillomavirus-associated (pre) neoplastic lesions. J Pathol 206:346–355PubMedCrossRefGoogle Scholar
  23. 23.
    Hubert P, van den Brule F, Giannini SL et al (1999) Colonization of in vitro-formed cervical human papillomavirus-associated (pre) neoplastic lesions with dendritic cells—role of granulocyte/macrophage colony-stimulating factor. Am J Pathol 154:775–784PubMedGoogle Scholar
  24. 24.
    Izadpanah A, Dwinell MB, Eckmann L et al (2001) Regulated MIP-3alpha/CCL20 production by human intestinal epithelium: mechanism for modulating mucosal immunity. Am J Physiol Gastrointest Liver Physiol 280:G710–G719PubMedGoogle Scholar
  25. 25.
    Jiang M, Milner J (2002) Selective silencing of viral gene expression in HPV-positive human cervical carcinoma cells treated with siRNA, a primer of RNA interference. Oncogene 21:6041–6048PubMedCrossRefGoogle Scholar
  26. 26.
    Jimenez-Flores R, Mendez-Cruz RF, Ojeda-Ortiz JF et al (2005) High-risk human papilloma virus infection decreases the frequency of dendritic Langerhans’ cells in the human female genital tract. Immunology 117:220–228CrossRefGoogle Scholar
  27. 27.
    Karin M, Ben-Neriah Y (2000) Phosphorylation meets ubiquitination: the control of NF-(kappa)B activity. Annu Rev Immunol 18:621–663PubMedCrossRefGoogle Scholar
  28. 28.
    Kimsey TF, Campbell AS, Albo D et al (2004) Co-localization of macrophage inflammatory protein-3alpha (Mip-3alpha) and its receptor, CCR6, promotes pancreatic cancer cell invasion. Cancer J 10:374–380PubMedCrossRefGoogle Scholar
  29. 29.
    Kleine-Lowinski K, Rheinwald JG, Fichorova RN et al (2003) Selective suppression of monocyte chemoattractant protein-1 expression by human papillomavirus E6 and E7 oncoproteins in human cervical epithelial and epidermal cells. Int J Cancer 107:407–415PubMedCrossRefGoogle Scholar
  30. 30.
    Matthews K, Leong CM, Baxter L et al (2003) Depletion of Langerhans cells in human papillomavirus type 16-infected skin is associated with E6-mediated down regulation of E-cadherin. J Virol 77:8378–8385PubMedCrossRefGoogle Scholar
  31. 31.
    McMurray HR, McCance DJ (2004) Degradation of p53, not telomerase activation, by E6 is required for bypass of crisis and immortalization by human papillomavirus type 16 E6/E7. J Virol 78:5698–5706PubMedCrossRefGoogle Scholar
  32. 32.
    Merrick DT, Blanton RA, Gown AM et al (1992) Altered expression of proliferation and differentiation markers in human papillomavirus 16 and 18 immortalized epithelial cells grown in organotypic culture. Am J Pathol 140:167–177PubMedGoogle Scholar
  33. 33.
    Nakayama T, Fujisawa R, Yamada H et al (2001) Inducible expression of a CC chemokine liver- and activation-regulated chemokine (LARC)/macrophage inflammatory protein (MIP)-3 alpha/CCL20 by epidermal keratinocytes and its role in atopic dermatitis. Int Immunol 13:95–103PubMedCrossRefGoogle Scholar
  34. 34.
    Petry KU, Scheffel D, Bode U et al (1994) Cellular immunodeficiency enhances the progression of human papillomavirus-associated cervical lesions. Int J Cancer 57:836–840PubMedCrossRefGoogle Scholar
  35. 35.
    Rubie C, Oliveira-Frick V, Rau B et al (2006) Chemokine receptor CCR6 expression in colorectal liver metastasis. J Clin Oncol 24:5173–5174PubMedCrossRefGoogle Scholar
  36. 36.
    Scheffner M, Munger K, Byrne JC et al (1991) The state of the p53 and retinoblastoma genes in human cervical carcinoma cell lines. Proc Natl Acad Sci USA 88:5523–5527PubMedCrossRefGoogle Scholar
  37. 37.
    Schmuth M, Neyer S, Rainer C et al (2002) Expression of the C–C chemokine MIP-3 alpha/CCL20 in human epidermis with impaired permeability barrier function. Exp Dermatol 11:135–142PubMedCrossRefGoogle Scholar
  38. 38.
    Sheppard KA, Rose DW, Haque ZK et al (1999) Transcriptional activation by NF-kappaB requires multiple coactivators. Mol Cell Biol 19:6367–6378PubMedGoogle Scholar
  39. 39.
    Tang S, Tao M, McCoy JP Jr et al (2006) Short-term induction and long-term suppression of HPV16 oncogene silencing by RNA interference in cervical cancer cells. Oncogene 25:2094–2104PubMedCrossRefGoogle Scholar
  40. 40.
    Thomas M, Pim D, Banks L (1999) The role of the E6–p53 interaction in the molecular pathogenesis of HPV. Oncogene 18:7690–7700PubMedCrossRefGoogle Scholar
  41. 41.
    Viac J, Guerin-Reverchon I, Chardonnet Y et al (1990) Langerhans cells and epithelial-cell modifications in cervical intraepithelial neoplasia—correlation with human papillomavirus infection. Immunobiology 180:328–338PubMedGoogle Scholar
  42. 42.
    Yoshinouchi M, Yamada T, Kizaki M et al (2003) In vitro and in vivo growth suppression of human papillomavirus 16-positive cervical cancer cells by E6 siRNA. Mol Ther 8:762–768PubMedCrossRefGoogle Scholar
  43. 43.
    Zur Hausen H (1991) Human papillomaviruses in the pathogenesis of anogenital cancer. Virology 184:9–13PubMedCrossRefGoogle Scholar
  44. 44.
    Zur Hausen H (2000) Papillomaviruses causing cancer: evasion from host-cell control in early events in carcinogenesis. J Natl Cancer Inst 92:690–698PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Jean-Hubert Caberg
    • 1
  • Pascale Hubert
    • 1
  • Ludivine Herman
    • 1
  • Michael Herfs
    • 1
  • Patrick Roncarati
    • 1
  • Jacques Boniver
    • 1
  • Philippe Delvenne
    • 1
  1. 1.Department of Pathology, GIGA-Cancer, B35University of LiegeLiegeBelgium

Personalised recommendations