Study of Human Leukocyte Antigen-Cw in Egyptian Patients With Vitiligo

April 2015 | Volume 14 | Issue 4 | Original Article | 359 | Copyright © 2015

Hussein M. Hassab El Naby MD,a Mohamed R. Alnaggar MD,a Mahmoud F. Abdelhamid MD,b
Khadiga Alsaid MD,c Eslam M. Al Shawadfy MD,b and Mohamed L. Elsaie MD MBAb

aDepartment of Dermatology and Andrology, Al-Azhar University, Cairo, Egypt
bDepartment of Dermatology and Venereology Research, National Research Centre, Cairo, Egypt
cDepartment of Molecular Genetics, National Research Centre, Cairo, Egypt

Abstract

Human leukocyte antigen (HLA) antigens vary considerably in different racial groups, and an analysis of results from several geographical locations suggests that vitiligo appears to be associated with different HLA antigens in different groups. The aim of this work was to assess the association of HLA-Cw with vitiligo in the Egyptian population. Forty unrelated patients with nonsegmental vitiligo and 20 matched controls were selected. A polymerase chain reaction sequence specific primer (PCR-SSP) method was used to determine HLA DNA typing. There was a statistically significant difference in the association of HLA-Cw6 with vitiligo in the 2 studied groups. A comparatively increased number of patients showed HLA-Cw2 and HLA-Cw7 (13.64%). However, there were no statistically significant differences. To the best of our knowledge, this is the first molecular study of HLA typing in Egyptian patients with vitiligo. Our findings are in agreement with earlier studies that reported statistically increased frequencies for allele of HLA-Cw6 in Northern Italian, Kuwaiti, Chinese Han, and Saudi populations (45.45%, P<.05).

J Drugs Dermatol. 2015;14(4):359-364.

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INTRODUCTION

Vitiligo can be a psychologically devastating disease, especially in darker skinned individuals. Vitiligo is a heritable condition. Although the inheritance pattern has not been fully established, up to 30% of patients report vitiligo in another family member, and up to 21% of first-generation family members may be affected. Offspring are at greatest risk, followed by siblings, parents, and grandparents. Vitiligo has been observed in monozygotic twins; the age of onset, extent, and course may be similar. Vitiligo affecting only 1 twin has also been reported.1 Vitiligo appears to be equally prevalent among men and women, with no difference in occurrence rates among skin type or race.

Vitiligo appears to be transmitted genetically in a polygenic/ multifactorial manner. The actual pathogenesis is under debate. 2 Human leukocyte antigen (HLA) complex is composed of several closely linked loci, each containing several alleles; it is the most gene-dense, polymorphic region of the human genome and is known to contain over 224 identified gene loci. Its inheritable nature and its frequent association with autoimmune diseases have prompted studies on the association of HLA with vitiligo. The associations of several HLA class I and class II antigens susceptible to the development of vitiligo have been investigated in different ethnic populations. Using linkage disequilibrium analysis in familial vitiligo, a study suggested that a major determinant of vitiligo might be located in HLA.3

The increased frequencies of HLA-A2, HLA-A30, HLA-Bw60, HLACw4, HLA-Cw6, HLA-Cw7, HLA-DR1, and HLA-DR4 in patients with vitiligo have been reported by several investigators.4,5,6,7,8 The aim of this work was to assess the association of HLA-Cw with vitiligo among the Egyptian population because to the best of our knowledge there is no published molecular study on the HLA antigens associated with vitiligo in this population.

SUBJECTS AND METHODS

Forty unrelated patients with nonsegmental vitiligo were selected and subjected to a full history and clinical examination, including the type and site of vitiliginous lesions. Twenty matched controls were selected randomly from disease-free, unrelated, apparently healthy donors.

HLA DNA Typing

DNA extraction was carried out with the classical phenol chloroform method from fresh whole blood samples that were anticoagulated by ethylenediaminetetraacetic acid (EDTA). A polymerase chain reaction sequence specific primer (PCR-SSP) method was used to determine HLA DNA typing. A dried primer stock solution consisting of an HLA-specific primer mix, ie, allele, and group specific was aliquoted in 0.2 mL PCR tubes.

The PCR master mix contained: 0.4 U/μL Taq Polymerase, 200 μM dNTPs, 50 mM KCI, 1.5 mM MgCl2, 10 mM Tris-HCI, pH 8.3, 0.001% w/v gelatin, 5% glycerol, and 100 μg/mL Cresol Red dye

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