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Allele and Haplotype Frequencies for HLA-DQ in Iranian Celiac Disease Patients

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Allele and Haplotype Frequencies for HLA-DQ in Iranian Celiac Disease Patients Submit a Manuscript: http://www.wjgnet.com/esps/ World J Gastroenterol 2014 May 28; 20(20): 6302-6308 Help Desk: http://www.wjgnet.com/esps/helpdesk.aspx ISSN 1007-9327 (print) ISSN 2219-2840 (online) DOI: 10.3748/wjg.v20.i20.6302 © 2014 Baishideng Publishing Group Inc. All rights reserved. CASE CONTROLBRIEF ARTICLE STUDY Allele and haplotype frequencies for HLA-DQ in Iranian celiac disease patients Mohammad Rostami-Nejad, Jihane Romanos, Kamran Rostami, Azita Ganji, Mohammad Javad Ehsani-Ardakani, Ali-Reza Bakhshipour, Homayoun Zojaji, Seyed Reza Mohebbi, Mohammad-Reza Zali, Cisca Wijmenga Mohammad Rostami-Nejad, Mohammad Javad Ehsani- antigen (HLA)-DQ2 and -DQ8 in Iranian celiac disease Ardakani, Homayoun Zojaji, Seyed Reza Mohebbi, Moham- (CD) patients and compare them to healthy Iranian mad-Reza Zali, Department of Celiac Disease, Gastroenterology controls. and Liver Diseases Research Centers, Shahid Beheshti University of Medical Sciences, Tehran 1985717411, Iran METHODS: To predict the HLA-DQA1 and -DQB1 Jihane Romanos, Cisca Wijmenga, Department of Genetics, genes, we used six previously reported HLA-tagging University of Groningen, University Medical Centre of Gronin- gen, 9700 RB Groningen, The Netherlands single nucleotide polymorphism to determine HLA Kamran Rostami, Department of Gastroenterology, Worcester- genotypes in 59 Iranian patients with ‘biopsy-confirmed’ shire Royal Hospital, Worcester, WR5 1DD, United Kingdom CD and in 151 healthy Iranian individuals. To test the Azita Ganji, Department of Gastroenterology, Imam Reza Hospi- transferability of the method, 50 cases and controls tal Mashhad University of Medical Sciences, Mashhad 1351143, were also typed using a commercial kit that identifies Iran individual carriers of DQ2, DQ7 and DQ8 alleles. Ali-Reza Bakhshipour, Department of Gastroenterology, Zahe- dan University of Medical Sciences, Zahedan 2143453, Iran RESULTS: In this pilot study 97% of CD cases (n = Jihane Romanos, Department of Genetics, Institute of Human Genetics, Lebanese American University, Byblos 36, Lebanon 57) and 58% of controls (n = 87) were carriers of HLA- Author contributions: Rostami-Nejad M contributed to sample DQ2 and/or HLA-DQ8 heterodimers, either in the ho- collection, article writing, final approval; Romanos J contributed mozygous or heterozygous state. The HLA-DQ pattern to HLA typing and drafting the article; Rostami K contributed of these 57 CD patients: heterozygous DQ2.2 (n = 14) to supervisor of this project and editing the manuscript; Ganji A and homozygous DQ2.2 (n = 1), heterozygous DQ2.5 (n contributed to sample collection from Mashhad city; Mohebbi SR = 33) and homozygous DQ2.5 (n = 8), heterozygous contributed to DNA extraction and analysis and interpretation of DQ8 (n = 13) and homozygous DQ8 (n = 2). Two CD data; Zojaji H contributed to Sample collection from Tehran city; patients were negative for both DQ2 and DQ8 (3%). Bakhshipour AR contributed to sample collection from Zahedan city; Zali MR and Ehsani-Ardakani MJ contributed to concept and design of the work, final approval; Wijmenga C contributed CONCLUSION: The prevalence of DQ8 in our CD pop- to supervising HLA typing, article writing, final approval. ulation was higher than that reported in other popula- Correspondence to: Mohammad Javad Ehsani-Ardakani, tions (25.4%). As reported in other populations, our MD, Department of Celiac Disease, Gastroenterology and Liver results underline the primary importance of HLA-DQ Diseases Research Centers, Shahid Beheshti University of Medi- alleles in the Iranian population’s susceptibility to CD. cal Sciences, Velenjak St., Shahid Chamran Highway, Tehran 1985717411, Iran. [email protected] © 2014 Baishideng Publishing Group Inc. All rights reserved. Telephone: +98-21-22432518 Fax: +98-21-22432517 Received: November 2, 2013 Revised: December 31, 2013 Key words: Human leukocyte antigen typing; Valida- Accepted: January 20, 2014 Published online: May 28, 2014 tion; Susceptibility; Celiac disease; Iran Core tip: We describe, for the first time, the distribution of human leukocyte antigen (HLA)-DQ2/DQ8 alleles in Iranian celiac disease patients compared to healthy Ira- Abstract nian controls. To assess this distribution we applied a AIM: To assess the distribution of human leukocyte single nucleotide polymorphism-based approach, which WJG|www.wjgnet.com 6302 May 28, 2014|Volume 20|Issue 20| Rostami-Nejad M et al . HLA typing in Iranian celiac disease patients was developed in European (Caucasian) study samples. large country. Based on previous studies in Iran, the most We also demonstrate the transferability of such an ap- common HLA haplotype in the different ethnic groups proach to the Iranian population. Our results underline is DQ2 and DQ7, with reported prevalences of 22.1% the primary importance of HLA-DQ alleles in the Iranian and 25%, respectively[14-17] (Figure 1). These studies were population’s susceptibility to celiac disease. performed in healthy populations in different parts of the country and found considerable similarities in the distribution of HLA class Ⅱ haplotypes with European Rostami-Nejad M, Romanos J, Rostami K, Ganji A, Ehsani- countries. However, Iranian CD patients have never been Ardakani MJ, Bakhshipour AR, Zojaji H, Mohebbi SR, Zali studied for HLA. MR, Wijmenga C. Allele and haplotype frequencies for HLA- Recently, Monsuur et al[13] described an HLA-tagging DQ in Iranian celiac disease patients. World J Gastroenterol single nucleotide polymorphism (SNP) method for de- 2014; 20(20): 6302-6308 Available from: URL: http://www. tecting the HLA risk alleles for CD. Using six SNPs, wjgnet.com/1007-9327/full/v20/i20/6302.htm DOI: http://dx.doi. HLA genotyping for specific CD genotypes could be per- org/10.3748/wjg.v20.i20.6302 formed in a high-throughput mode. The results of Mon- suur et al[13] and Koskinen et al[12] showed that the sensi- tivity and specificity of this test in recognizing DQ2.2, DQ2.5, DQ7, and DQ8 haplotypes was > 99% in the INTRODUCTION Dutch population, and > 98% in the United Kingdom, Celiac disease (CD) is characterized by malabsorption of Spanish, Finnish, Hungarian, and Italian populations. nutrients in the small intestine after ingestion of wheat The main aim of this study was to provide new infor- gluten or related proteins from rye and barley. The dis- mation on HLA-DQ associations with CD in the Iranian ease is characterized by villous atrophy of the small intes- patients, which has never been investigated before, by us- tinal mucosa. With adherence to a strict gluten-free diet, ing the tagging SNP method. most CD patients show prompt clinical and histological improvement, with relapse of the symptoms if gluten is [1-3] MATERIALS AND METHODS re-introduced . Similar to Western studies, a high preva- lence of CD has been reported in Iranian general popula- Patients tion (about 1%)[3]. A total of 59 CD patients (21 males and 38 females, me- Many studies reflect the importance of genetic factors dian age 30.5 years, range 7-67 years) were included: 12 in the pathogenesis of CD[3]. A recent study by Anderson patients from East Iran (Zahedan Province, Baloch eth- et al[4] suggested that a combination of human leukocyte nicity), 13 from North Iran (Mashhad Province, Fars), 10 antigen (HLA) typing and confirmatory serology could from West Iran (Lorestan and Kurdistan Provinces, Lour decrease the number of unnecessary endoscopies. and Kurd ethnic groups), and 24 patients from Tehran, in The HLA-DQ2 heterodimer, which is coded by al- the center of Iran (Fars). All patients had positive tTGA leles DQA1*05 and DQB1*02, is present in the vast and/or EMA antibodies and histology according to the majority of CD patients including approximately 95% of Rostami-Marsh classification (Marsh Ⅰ-Ⅲc)[18]. CD patients of Northern-European descent[4]. The most important risk factor for CD is the DQ2.5 haplotype[4]. Controls Alpha and beta chains of this heterodimer are encoded A total of 151 healthy controls (78 females and 73 males, together in cis (DQA1*05 and DQB1*02) on a DRB1*03 median age 32.7 years, range 5-83 years) were selected haplotype. Other DQ heterodimers that are encoded in from those who were negative for CD serological screen- trans on DR7 haplotypes are related to the DQ2.2 hap- ing in the study areas (25 from Zahedan Province, 30 lotype (DQA1*0201/DQB1*02)[5-13]. Those CD patients from Mashhad, 26 from Lorestan and Kurdistan prov- who do not carry the DQ2 heterodimer, and are encoded inces, and 70 from Tehran) and matched by age and sex. by the DR4 haplotype, carry HLA-DQ8 (a1*0301, None of them had a personal or family history of cancer β1*0302)[4]. HLA-DQ2 is common in Europeans and is or autoimmune diseases. The study was approved by the expressed in 25%-30% of the healthy European popula- ethical committees of the Gastroenterology and Liver tion. Most of the CD patients who do not carry either Diseases Research Centers, Shahid Beheshti University DQ2.5 or DQ8, carry half of the DQ2.5 or DQ2.2 mol- of Medical Sciences, Tehran, and all the participants (or ecule (that is either HLA-DQA1*05 or HLA-DQB1*02), their parents/guardians) were informed about the study suggesting that carrying even some of the risk molecules according to the study protocol and gave their written still has functional implications for the risk of CD[13]. The informed consent. majority of studies of HLA in CD are focused on Cau- casian populations and detailed information on patient HLA typing cohorts from diverse ethnic groups is lacking. DNA was extracted using the phenol chloroform meth- There are a number of ethnic groups in Iran, in- od[19]. The six HLA-tagging SNPs reported by Monsuur cluding Persians (Fars), Kurds, Lours, Arabs, Turkmen, et al[13] were genotyped using 5 ng of DNA from the Ira- Balochs, and Turks. Most Iranians are Muslims but Zoro- nian participants, and using TaqMan chemistry and cus- astrians, Jews, Armenians, and Nestorians also live in this tom assays from Applied Biosystems (ABI, Foster City, WJG|www.wjgnet.com 6303 May 28, 2014|Volume 20|Issue 20| Rostami-Nejad M et al .
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