HLA-DRB 1 * 03 And DRB 1 * 15 Frequency In Helicobacter Pylori Superficial Gastritis

Al-Kindy College Medical Journal 2017: Vol.13 No. 2 63 en.jkmc.uobaghdad.edu.iq ABSTRACT Background: Background: Helicobacter pylori is an important gastrointestinal bacteria related to the development of superficial atrophic gastritis, peptic ulcer and gastric cancer. Human leukocyte antigens (HLA) may play an important roles in host immune responses to H pylori antigens. Aim of the study: to investigate the association between HLA-DRB1 genotypes and superficial gastritis with H. pylori infection in an Iraqi patients. Patients and methods: Sixty patients with superficial gastritis and 100 individuals with apparently normal results after endoscopic examination were recruited from Al-Kindy Teaching Hospital Gastrocolonoscope Unit between January 2014 and July 2016. All study patients and controls group underwent upper gastrointestinal endoscopic examinations. They were analyzed for CagA antibodies Immunoglobulin G (IgG) for H. pylori and HLA Class II genotyping (HLA-DRB1). Results: Patients with superficial gastritis with H pylori. The infection showed significant expression (P=0.0001) DRB1*03:01 genotypes and DRB1*15:01 (P= 0.004) as compared to control group. Conclusions: Genetic factor may play a role in gastritis. HLA-DRB1*03 and HLA-DRB1*15 may increased the susceptibility to superficial gastritis in H pylori infected patients.

astritis is the term of inflammation of stomach walls inflammation to various degrees.This is either including the superficial inner lining of the stomach (mucosa) or it can extend through the entire wall of the stomach (1).Persistent inflammation of the gastric mucosa leads to gastritis caused by many external different factors like H.pylori (Helicobacter pylori) infection or the excessive and/or long term use of medication suchas NSAIDs (non-steroidal antiinflammatory drugs) (2).However, anyone can get gastritis; but there are certain factors can raise the risk.These include: weakened immune systems for example immune-mediated reaction and is known as autoimmune atrophic gastritis (3) and genetic factors that predispose them to developing gastritis (4).Gastritis might be associated with polymorphisms of specific allelic that involved in specific inflammatory response genes (interleukin-1 gene cluster and Toll-like receptors (TLRs) (5)., detoxification enzymes, and cancer-related processes like single A/T SNP at position -251 from the transcription start site in the promoter region of the IL-8 gene (6).Consequently polymorphisms in inflammationrelated genes are factors important in the development of gastroduodenal diseases in Helicobacter pyloriinfected individuals.Other important polymorphic genetic factor is Human Leukocyte Antigen (HLA) which is play particular roles in host immune responses to bacterial antigens of H pylori (7).Class II molecules of HLA is a hetrodimer membrane glycoproteins expressed on the surface of antigen presenting cells (8).It presents exogenous antigenic peptide to T helper cells, so the interaction between T cell receptors, antigenic peptide and MHC molecule determines T cell activation and an immune response to various antigens (9).
It was reported that there is an association of Class II (HLA-DQA1 and DQB1)alleles with gastric disease (10).The present study was done in order to investigate the association of HLA-DRB1 genetic diversity in individuals infected with H. pylori in an Iraqi Arab Muslims population.
Methods: Patients with superficial gastritis after gastroscope examination were done to them and individuals with apparently normal results after endoscopic examination were recruited from Al-Kindy Teaching Hospital -Gastrocolonoscope Unit between January 2014 and July 2016.Written informed consent was obtained from all patients with superficial gastritis and control group for this study.The study protocol was reviewed and approved by the Scientific and Ethical Committee of Al-kindy medical college and Al-Kindy Teaching Hospital.The patient and control groups were in the same aged for both sexes.In situe hybridization using Locus-and allelesspecific amplification of genomic patients and control DNA was performed for DRB1.DNA Amplification and Hybridization from both groups were performed using a sequence-specific oligonucleotide probes (SSOP) by HLA-DRB1 amplification and hybridization kits (SSO HLA type DRB1 plus and Mastermix for HLA type DRB1 Amp plus kits -Innogenetics-Belgium) by AutoLipa -48Innogenetics-Belgum.The results were interpreted using LiRas version-5.0software-Innogenetics-Belgium.

Statistical Analysis:
HLA-DRB1 frequencies were determined by direct counting.The frequency of each allele was compared between two groups using chisquare test fisher exact test using MiniTab version.3.0 software.In each comparison, the Odds ratio (OR) along with the 95% confidence interval (95% CI) was calculated.Gene frequencies for both groups wereestimated.P-value less than 0.05 were considered statistically significant.
Results : A total of 60 patients with superficial gastritis were evaluated, together with 100 control group.The mean age of patients was 44.37 ±11.54, as compared with 46 ± 10.12 for the controls .The males were 50% and the rest was females.The male to female sex ratio in the control group was 1:1 There is a significant increase of H. pylori infection (p=0.0001) in superficial gastritis patients than control group.The Odd ratio (OD)= 18.878 with 95% CI= from    (15).In Lombok Indonesian patients, HLADQB1*0401 gene play important roles in H. pylori infection, but there was no statistically significant association between HLA-DQA1 or DQB1 haplotypes and H.pylori infection (16).These differences with our study may be related to race of the studying patients, religion, sample size, criteria of patient's selection and method used in detection associated alleles.Thus, early detection of atrophic gastritis and gastric cancer can be done using multiple markers like HLA-DR, HLA-DQ, age, sex and H pylori infection (17).Other studies done in Iraq showed that there is an association between HLA and different diseases in Iraq (18) like DR3 and DR4 is common in Insulin dependent diabetes mellitus (19,20), Inflammatory bowel disease was associated with DR8 (21).In conclusion, our results, together with other studies , support that genetic constitution through HLA-DR locus determines the mechanism and pathogenesis of disease as well as clinical and pathologic outcomes, triggered by the interaction between environmental factors and genetic factors.In other words, immunogenetic background among different population with different ethnicities is manifested as resistance or susceptibility to the development of superficial gastritis.
8.190-43.513.The relative risk =6.363 that indicates an association between H. pylori and superficial gastritis disease as shown in table-1-.The observed and expected phenotypes of all alleles for the patients group as demonstrated in table-2-were in a good agreement with Hardy-Weinberg equilibrium as shown in table-3-.The frequencies of HLA*DRB1 were investigated in the control and patients groups of Iraqi Arab Muslims and analyzed for identifying the DRB1* alleles using DNA based methodology (PCR-SSOP).

Table - 3
-Hardy -Weinberg equilibrium in HLA-DRB1 locus of patients with superficial gastritis disease.

Table - 4
-Frequencies of HLA-DRB1 alleles in patients with superficial gastritis disease compared with control group.
0.011 en.jkmc.uobaghdad.edu.iqFigure-1-Results of DNA electrophoresis after amplification Discussion: The current study investigated association between patients of superficial gastritis (with an H pylori infection) and HLA-DRB1 alleles in Iraqi Arab Muslims.*0501 in Mexican Mestizo patients with gastric carcinoma in comparison with control group (P = 1 x 10(-6), OR = 13.07;95% CI= 2.82-85.14)(14).The authors have reported that human leukocyte antigen (HLA)-DQB1*0401 plays an important role in the development of atrophic gastritis in H. pylori infected patients and it is a useful marker for determining susceptibility to intestinal type gastric cancer