Detection of auto antibodies and transplantation of cultured autologous melanocytes for the treatment of vitiligo

Zhu,Mei‑Cai; Ma,Hong‑Yu; Zhan,Zhi; Liu,Cheng‑Gang; Luo,Wei; Zhao,Guang

doi:10.3892/etm.2016.3949

January-2017 Volume 13 Issue 1

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January-2017 Volume 13 Issue 1

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Article Open Access

Detection of auto antibodies and transplantation of cultured autologous melanocytes for the treatment of vitiligo

Authors:
- Mei‑Cai Zhu
- Hong‑Yu Ma
- Zhi Zhan
- Cheng‑Gang Liu
- Wei Luo
- Guang Zhao
View Affiliations / Copyright

Affiliations: Center of Clinical Laboratory, Air Force General Hospital, Beijing 100036, P.R. China, Department of Dermatology, Air Force General Hospital, Beijing 100036, P.R. China

Copyright: © Zhu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Pages: 23-28
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Published online on: December 2, 2016

https://doi.org/10.3892/etm.2016.3949
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Abstract

The aim of the present study was to establish an immunofluorescence method of antibody detection to identify melanocytes in the serum of vitiligo patients. Furthermore, we aimed to establish a method for the culture and proliferation of autologous pure melanocytes and to observe the effect of their transplantation for the treatment of vitiligo. Suspension of epidermal cells with melanocytes was performed using trypsin digestion of normal epiderm from eyelid operation and melanocytes were selectively cultured and proliferated in serum-free M2 medium. FITC-labeled rabbit anti‑-human antibody was used to detect the relative fluorescence intensity of the melanocytes. After identification with immunological and biological examinations, the melanocytes were transplanted to depigmented areas of vitiligo. Repigmentation was observed continuously. The results indicated that melanocytes could be selectively proliferated in the medium. Subsequently, pure melanocytes without contamination of fibroblast and keratinocyte were harvested. A total of 34 patients suffering vitiligo for between 3 months and 20 years with depigmented area (between 4 cm2 and 70% of body surface) were divided into 19 cases of developing stage and 15 cases of stable stage, according to the change of depigmentation. A total of 15 developing cases were positive for the antibody against melanocytes, with the positive rate of 79%. The titers of serum was >1:50 in 10 patients at the developing stage, and 5 developing patients were 1:10. Among the 15 stable cases, four were positive, with a positive rate of 27%. Fluorescence of antibody was localized in the cytoplasm of the melanocytes. Autologous melanocytes of vitiligo patients could be selectively proliferated in the medium. Next, pure melanocytes without contamination with fibroblasts and keratinocytes were harvested. A total of 16 vitiligo patients with 28 depigmented areas (2‑200 cm2) were treated with transplantation of melanocytes. Repigmentation of the transplanted areas appeared as red coloration after one month. All the vitiligous areas received transplantation were repigmented significantly with hypo‑ or hyper‑pigmentation after 3‑5 months. After 6‑8 months, 87.5% of lesions showed repigmentation of >50% of the lesion area. No scarring or other side‑effects occurred. After follow‑up of 5 years, no relapse was observed in transplantation area. Thus, an immunofluorescence method for the test of antibody to melanocytes in the serum of vitiligo patients was established. Transplantation of cultured autologous melanocytes was an effective and safe measure for treatment of vitiligo, particularly for patients with a large depigmented area.

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1	Lu T, Gao TT, Wang AH, Li Q, Li CY, Zhao XD and Sun DJ: The prevalence of vitiligo in Shaanxi Province of China. Zhong Hua Pi Fu Ke Za Zhi. 37:406–407. 2004.(In Chinese).
2	Namazi MR: Neurogenic dysregulation, oxdative stress, autoimmunity and melanocytorrhagy in vitiligo: Can they be interconnected? Pigment Cell Res. 20:360–363. 2007. View Article : Google Scholar : PubMed/NCBI
3	Cui J, Arita Y and Bystryn JC: Cytolytic antibodies to melanocytes in vitiligo. J Invest Dermatol. 100:812–825. 1993. View Article : Google Scholar : PubMed/NCBI
4	Ruiz-Argüelles A, Brito GJ, Reyes-Izquierdo P, Pérez-Romano B and Sánchez-Sosa S: Apoptosis of melanocytes in vitiligo results from antibody penetration. J Autoimmun. 29:281–286. 2007. View Article : Google Scholar : PubMed/NCBI
5	Sandoval-Cruz M, García-Carrasco M, Sánchez-Porras R, Mendoza-Pinto C, Jiménez-Hernández M, Munguía-Realpozo P and Ruiz-Argüelles A: Immunopathogenesis of vitiligo. Autoimmun Rev. 10:762–765. 2011. View Article : Google Scholar : PubMed/NCBI
6	Horikawa T, Mishima Y, Nishino K and Ichihashi M: Horizontal and vertical pigment spread into surrounding piebald epidermis and hair follicles after suction blister epidermal grafting. Pigment Cell Res. 12:175–180. 1999. View Article : Google Scholar : PubMed/NCBI
7	Boersma BR, Westerhof W and Bos JD: Repigmentation in vitiligo vulgaris by autologous minigrafting: Results in nineteen patients. J Am Acad Dermatol. 33:990–995. 1995. View Article : Google Scholar : PubMed/NCBI
8	Lu T, Gao TW, Liu YF, Li CY and Sun LC: The explore of allogenic transplantation of melanocytes to treat vitiligo. Zhong Guo Pi Pu Xing Bing Xue Za Zhi. 15:240–242. 2001.(In Chinese).
9	Mulekar SV: Melanocyte-keratinocyte cell transplantation for stable vitiligo. Int J Dermatol. 42:132–136. 2003. View Article : Google Scholar : PubMed/NCBI
10	Chen YF, Chang JS, Yang PY, Hung CM, Huang MH and Hu DN: Transplant of cultured autologous pure melanocytes after laser-abrasion for the treatment of segmental vitiligo. J Dermatol. 27:434–439. 2000. View Article : Google Scholar : PubMed/NCBI
11	Olsson MJ and Juhlin L: Long-term follow-up of leucoderma patients treated with transplants of autologous cultured melanocytes, ultrathin epidermal sheets and basal cell layer suspension. Br J Dermatol. 147:893–904. 2002. View Article : Google Scholar : PubMed/NCBI
12	Olsson MJ and Juhlin L: Leucoderma treated by transplantation of a basal cell layer enriched suspension. Br J Dermatol. 138:644–648. 1998. View Article : Google Scholar : PubMed/NCBI
13	Beck AJ, Phillips J, Smith-Thomas L, Short RD and MacNeil S: Development of a plasma-polymerized surface suitable for the transplantation of keratinocyte-melanocyte cocultures for patients with vitiligo. Tissue Eng. 9:1123–1131. 2003. View Article : Google Scholar : PubMed/NCBI
14	Le-Poole IC and Luiten RM: Autoimmune etiology of generalized vitiligo. Curr Dir Autoimmun. 10:227–243. 2008. View Article : Google Scholar : PubMed/NCBI
15	Kemp EH, Waterman EA, Hawes BE, O'Neill K, Gottumukkala RV, Gawkrodger DJ, Weetman AP and Watson PF: 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
16	Kao CH, Ko WC, Ko SS and Tsai RY: A comparative study on autologous graft for segmental type vitiligo. Dermatol Sinica. 13:65–74. 1995.
17	Halaban R and Alfano FD: Selective elimination of fibroblasts from cultures of normal human melanocytes. In Vitro. 20:447–450. 1984. View Article : Google Scholar : PubMed/NCBI
18	Phillips J, Gawkrodger DJ, Caddy CM, Hedley S, Dawson RA, Smith-Thomas L, Freedlander E and Mac Neil S: Keratinocytes suppress TRP-1 expression and reduce cell number of co-cultured melanocytes: Implications for grafting of patients with vitiligo. Pigment Cell Res. 14:116–125. 2001. View Article : Google Scholar : PubMed/NCBI
19	Eisinger M and Mark O: Selective proliferation of normal human melanocytes in the presence of phorbol ester and cholera toxin. Proc Natl Acad Sci USA. 79:2018–2022. 1982. View Article : Google Scholar : PubMed/NCBI

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Detection of auto antibodies and transplantation of cultured autologous melanocytes for the treatment of vitiligo

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