CD8+ T cells from vitiligo perilesional margins induce autologous melanocyte apoptosis

Wu,Jilong; Zhou,Miaoni; Wan,Yinsheng; Xu,Aie

doi:10.3892/mmr.2012.1117

CD8+ T cells from vitiligo perilesional margins induce autologous melanocyte apoptosis

Authors:
- Jilong Wu
- Miaoni Zhou
- Yinsheng Wan
- Aie Xu
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Affiliations: Department of Dermatology, The Third People's Hospital of Hangzhou, Hangzhou 310009, P.R. China, Department of Biology, Providence College, Providence, RI 02918, USA
Published online on: October 8, 2012 https://doi.org/10.3892/mmr.2012.1117
Pages: 237-241

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Abstract

Cell-mediated autoimmunity has been suggested to be involved in the melanocyte apoptosis that occurs in vitiligo. We investigated the cytotoxicity to autologous melanocytes of CD8+ T cells from the perilesional margins and peripheral blood samples of vitiligo patients. CD8+ T cells isolated from skin biopsied from the edges of depigmented skin patches of vitiligo patients or from peripheral blood samples of the same donors were proliferated in culture medium. The primary cultures of CD8+ T cells and autologous melanocytes were mixed at ratios of 1:1, 1:2 or 1:5 and incubated for 3 days. The apoptosis of the melanocytes was analyzed by flow cytometry. Secreted cytokines in selected samples were measured by cytokine arrays. The results show that the CD8+ T cells were successfully isolated from the vitiligo perilesional margins. This cell population showed a significantly higher percentage of CD69 expression (56.13±3.55 versus 29.93±2.35%, p<0.01) and CD137 expression (41.74±1.06 versus 25.97±1.63%, p<0.01) compared with CD8+ T cells in peripheral blood from the same donors. The co-culturing of CD8+ T cells from lesional skin with autologous melanocytes induced apoptosis in the melanocytes (16.63±1.21, 16.71±0.63 and 18.32±1.60% for CD8+ T cells and autologous melanocytes at ratios of 1:1, 1:2 and 1:5, respectively). IL-6 levels were much higher in the co-culture (3.01-fold higher than in a melanocyte monoculture and 17.32-fold higher than in a CD8+ T-cell monoculture). The CD8+ T cells were also demonstrated to secrete more IL-13. Taken together, our data demonstrate that the infiltration of active CD8+ T cells takes place in the vitiligo perilesional margins. Those CD8+ T cells present significantly higher activation levels and higher cytotoxicity to autologous melanocytes than their counterparts from peripheral blood samples. These data suggest that CD8+ T cells are likely to be involved in the pathogenesis of vitiligo.

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1	Yee C, Thompson JA, Roche P, et al: Melanocyte destruction after antigen-specific immunotherapy of melanoma: direct evidence of T cell-mediated vitiligo. J Exp Med. 192:1637–44. 2000. View Article : Google Scholar : PubMed/NCBI
2	Kemp EH, Waterman EA, Hawes BE, et al: The melanin-concentrating hormone receptor 1, a novel target of autoantibody responses in vitiligo. J Clin Invest. 109:923–930. 2002. View Article : Google Scholar : PubMed/NCBI
3	Palermo B, Campanelli R, Garbelli S, et al: Specific cytotoxic T lymphocyte responses against Melan-A/MART1, tyrosinase and gp100 in vitiligo by the use of major histocompatibility complex/peptide tetramers: the role of cellular immunity in the etiopathogenesis of vitiligo. J Invest Dermatol. 117:326–332. 2001. View Article : Google Scholar
4	Glassman SJ: Vitiligo, reactive oxygen species and T-cells (Review). Clin Sci (Lond). 120:99–120. 2011. View Article : Google Scholar : PubMed/NCBI
5	Le Poole IC and Luiten RM: Autoimmune etiology of generalized vitiligo (Review). Curr Dir Autoimmun. 10:227–243. 2008.PubMed/NCBI
6	Le Poole IC, ElMasri WM, Denman CJ, et al: Langerhans cells and dendritic cells are cytotoxic towards HPV16 E6 and E7 expressing target cells. Cancer Immunol Immunother. 57:789–797. 2008.PubMed/NCBI
7	Ongenae K, Van Geel N and Naeyaert JM: Evidence for an autoimmune pathogenesis of vitiligo (Review). Pigment Cell Res. 16:90–100. 2003. View Article : Google Scholar : PubMed/NCBI
8	Badri AM, Todd PM, Garioch JJ, et al: An immunohistological study of cutaneous lymphocytes in vitiligo. J Pathol. 170:149–155. 1993. View Article : Google Scholar : PubMed/NCBI
9	Wankowicz-Kalinska A, van den Wijngaard RM, Tigges BJ, et al: Immunopolarization of CD4⁺ and CD8⁺ T cells to Type-1-like is associated with melanocyte loss in human vitiligo. Lab Invest. 83:683–695. 2003.
10	Wankowicz-Kalinska A, Le Poole C, van den Wijngaard R, et al: Melanocyte-specific immune response in melanoma and vitiligo: two faces of the same coin? Pigment Cell Res. 16:254–260. 2003. View Article : Google Scholar : PubMed/NCBI
11	van den Wijngaard RM, Aten J, Scheepmaker A, et al: Expression and modulation of apoptosis regulatory molecules in human melanocytes: significance in vitiligo. Br J Dermatol. 143:573–581. 2000.PubMed/NCBI
12	van den Wijngaard R, Wankowicz-Kalinska A, Le Poole C, et al: Local immune response in skin of generalized vitiligo patients. Destruction of melanocytes is associated with the prominent presence of CLA⁺ T cells at the perilesional site. Lab Invest. 80:1299–1309. 2000.PubMed/NCBI
13	Bos JD, Zonneveld I, Das PK, et al: The skin immune system (SIS): distribution and immunophenotype of lymphocyte subpopulations in normal human skin. J Invest Dermatol. 88:569–573. 1987. View Article : Google Scholar : PubMed/NCBI
14	Gross AJ, Hunt HH, Cantor AB and Clark BC: Sample size determination in clinical trials with an emphasis on exponentially distributed responses. Biometrics. 43:875–883. 1987. View Article : Google Scholar : PubMed/NCBI
15	van den Boorn JG, Konijnenberg D, Dellemijn TA, et al: Autoimmune destruction of skin melanocytes by perilesional T cells from vitiligo patients. J Invest Dermatol. 129:2220–2232. 2009.PubMed/NCBI
16	Hong WS, Hu DN, Qian GP, et al: Treatment of vitiligo in children and adolescents by autologous cultured pure melanocytes transplantation with comparison of efficacy to results in adults. J Eur Acad Dermatol Venereol. 25:538–543. 2011. View Article : Google Scholar : PubMed/NCBI
17	Hong WS, Hu DN, Qian GP, et al: Ratio of size of recipient and donor areas in treatment of vitiligo by autologous cultured melanocyte transplantation. Br J Dermatol. 165:520–525. 2011.PubMed/NCBI
18	Yu HS, Chang KL, Yu CL, et al: Alterations in IL-6, IL-8, GM-CSF, TNF-alpha, and IFN-gamma release by peripheral mononuclear cells in patients with active vitiligo. J Invest Dermatol. 108:527–529. 1997. View Article : Google Scholar : PubMed/NCBI
19	Moretti S, Spallanzani A, Amato L, et al: New insights into the pathogenesis of vitiligo: imbalance of epidermal cytokines at sites of lesions. Pigment Cell Res. 15:87–92. 2002. View Article : Google Scholar : PubMed/NCBI
20	Moretti S, Spallanzani A, Amato L, et al: Vitiligo and epidermal microenvironment: possible involvement of keratinocyte-derived cytokines. Arch Dermatol. 138:273–274. 2002. View Article : Google Scholar : PubMed/NCBI
21	Tu CX, Gu JS and Lin XR: Increased interleukin-6 and granulocyte-macrophage colony stimulating factor levels in the sera of patients with non-segmental vitiligo. J Dermatol Sci. 31:73–78. 2003. View Article : Google Scholar : PubMed/NCBI
22	Neurath MF and Finotto S: IL-6 signaling in autoimmunity, chronic inflammation and inflammation-associated cancer (Review). Cytokine Growth Factor Rev. 22:83–89. 2011. View Article : Google Scholar : PubMed/NCBI
23	Chalaris A, Garbers C, Rabe B, et al: The soluble Interleukin 6 receptor: generation and role in inflammation and cancer. Eur J Cell Biol. 90:484–494. 2011. View Article : Google Scholar : PubMed/NCBI
24	Krasagakis K, Hettmannsperger U, Tebbe B and Garbe C: Cutaneous metastatic angiosarcoma with a lethal outcome, following radiotherapy for a cervical carcinoma. Br J Dermatol. 133:610–614. 1995. View Article : Google Scholar : PubMed/NCBI
25	Krasagakis K, Garbe C, Zouboulis CC and Orfanos CE: Growth control of melanoma cells and melanocytes by cytokines. Recent Results Cancer Res. 139:169–182. 1995. View Article : Google Scholar : PubMed/NCBI
26	Krasagakis K, Garbe C, Eberle J and Orfanos CE: Tumour necrosis factors and several interleukins inhibit the growth and modulate the antigen expression of normal human melanocytes in vitro. Arch Dermatol Res. 287:259–265. 1995. View Article : Google Scholar : PubMed/NCBI
27	Kotobuki Y, Tanemura A, Yang L, et al: Dysregulation of melanocyte function by Th17-related cytokines: significance of Th17 cell infiltration in autoimmune vitiligo vulgaris. Pigment Cell Melanoma Res. 25:219–230. 2012. View Article : Google Scholar : PubMed/NCBI
28	Dienz O and Rincon M: The effects of IL-6 on CD4 T cell responses (Review). Clin Immunol. 130:27–33. 2009. View Article : Google Scholar : PubMed/NCBI
29	Dienz O, Eaton SM, Bond JP, et al: The induction of antibody production by IL-6 is indirectly mediated by IL-21 produced by CD4⁺ T cells. J Exp Med. 206:69–78. 2009. View Article : Google Scholar : PubMed/NCBI
30	Yoshida H, Hashizume M, Suzuki M and Mihara M: Anti-IL-6 receptor antibody suppressed T cell activation by inhibiting IL-2 production and inducing regulatory T cells. Eur J Pharmacol. 634:178–183. 2010. View Article : Google Scholar : PubMed/NCBI
31	Yoshida H, Hashizume M and Mihara M: IL-6 blockade preferentially inhibits Th17 differentiation in collagen-induced arthritis. Rheumatol Int. 31:127–131. 2011. View Article : Google Scholar : PubMed/NCBI
32	Romagnani S, Maggi E, Liotta F, et al: Properties and origin of human Th17 cells (Review). Mol Immunol. 47:3–7. 2009. View Article : Google Scholar
33	Suzuki T, Shoji S, Yamamoto K, et al: Essential roles of Lyn in fibronectin-mediated filamentous actin assembly and cell motility in mast cells. J Immunol. 161:3694–3701. 1998.PubMed/NCBI
34	Vercelli D: Genetics of IL-13 and functional relevance of IL-13 variants (Review). Curr Opin Allergy Clin Immunol. 2:389–393. 2002. View Article : Google Scholar : PubMed/NCBI
35	Mentink-Kane MM and Wynn TA: Opposing roles for IL-13 and IL-13 receptor alpha 2 in health and disease (Review). Immunol Rev. 202:191–202. 2004. View Article : Google Scholar : PubMed/NCBI
36	Mentink-Kane MM, Cheever AW, Thompson RW, et al: IL-13 receptor alpha 2 down-modulates granulomatous inflammation and prolongs host survival in schistosomiasis. Proc Natl Acad Sci USA. 101:586–590. 2004. View Article : Google Scholar : PubMed/NCBI