Determination of HLA-B27 subtypes in Iranian patients with ankylosing spondylitis

The human leukocyte antigen-B27 is one of the class I molecules of the major histocompatibility complex which is strongly associated with ankylosing spondylitis [AS]. The strength of the disease association with B27 varies markedly among racial and ethnic populations. It is an allele family, which constitutes about 31 subtypes, with a considerable geographic and ethnic difference in distribution. It is important to know whether certain subtypes show any preferential association with AS. Because there is no report regarding HLA-B27 subtypes in Iranian patients with AS, main purpose of the present study was to assess the frequency of subtypes of human leukocyte antigen [HLA]-B27 in patients with ankylosing spondylitis in Iranian population One hundred and nineteen AS patients [82 HLA-B27 positive and 37 HLA-B27 negative] were selected for this study. HLA-B27 positive patients were by polymerase chain reaction amplification with sequence-specific primers [PCR-SSP] for B 27 subtyping. The results of present study revealed that only two subtypes were detected in Iranian patients, including B 2705 [52 patients, 63.4%] and B 2702 [30 patients, 36.6%]. Our results showed a restricted number of HLA-B27 subtypes associated with AS in Iran and an elevated frequency of the B 2705 allele in these patients similar to other Euro-Caucasoid [Aryan] groups in the world

HLA class II allele and haplotype frequencies in Iranian patients with leukemia

Previous studies demonstrated significant differences in a number of HLA allele frequencies in leukemia patients and normal subjects. In this study, we have analyzed HLA class II alleles and haplotypes in 110 leukemia patients [60 acute myelogenous leukemia "AML", 50 chronic myelogenous leukemia"CML"] and 180 unrelated normal subjects. Blood samples were collected from all of the patients and control subjects. DNA was extracted by salting out method and HLA typing was performed using PCR-SSP method. Significant positive association with AML was obtained for HLA-DRB1*11allele [35% vs. 24.7%, P=0.033]. Two alleles including HLA-DRB4 and-DQB1*0303 were significantly less frequent in AML patients than in controls. HLA-DQB1*0303 allele was never observed in CML patients compared with allele frequency in controls [4.2%]. According to haplotype analysis, HLA-DRB1*0101/DQA1*0104/-DQB1*0501 frequencies were significantly higher and-DRB1*16/-DQA1*01021/-DQB1*0501 frequencies were significantly lower in CML patients than in controls .In conclusion it is suggested that HLA-DRB1*16 allele and HLA-DRB1*15/-DQA1*0103/-DQB1*06011 and-DRB1*16/-DQA1*01021/-DQB1*0501 haplotypes predispose individuals to AML and HLA-DRB4 allele predispose to CML. Future studies are needed to confirm these results and establish the role of these associations in AML and CML

HLA class II allele and haplotype frequencies in Iranian patients with acute myelogenous leukemia and control group

Previous studies have demonstrated some significant differences in HLA allele frequencies in leukemic patients and normal subjects. We have analyzed HLA class II alleles and haplotypes in 60 Iranian patients with acute myelogenous leukemia [AML] and 180 unrelated normal subjects. Blood samples were collected after obtaining informed consents. From the patients and control DNA extraction and HLA typing were performed using PCR-SSP method. Significant positive association with the disease was found for HLA-DRB1*11 allele [35% vs. 24.7%, p=0.033]. Two alleles including HLA-DRB4 and -DQB1*0303 were found to be significantly decreased in patients compared to controls. Regarding haplotype analysis, no significant association was found between case and control groups. It is suggested that HLA-DRB1*11 allele plays as a presumptive predisposing factor while the HLA-DRB4 and -DQB1*0303 alleles are suggested as protective genetic factors against acute myelogenous leukemia. Larger studies are needed to confirm and establish the role of these associations with acute myelogenous leukemia

Cytokine gene polymorphisms in iranian patients with beta-thalassemia major

beta-thalassemia as a hereditary disease is defined as defective synthesis of beta -globin chains, resulting in erythropoiesis abnormalities and severe anemia. Different studies have shown that cytokines and cytokine gene polymorphisms play a major role in the pathogenesis of beta -thalassemia. Single nucleotide polymorphisms [SNPs] within the promoter region or other regulatory sequences of cytokine genes lead to overall production of cytokines. To analyze the genetic profile of Thl and Th2 cytokines in Iranian patients with beta -thalassemia major. Allelic and genotype frequencies of cytokine genes were determined in 30 thalassemia patients and 40 healthy subjects using PCR-SSP assay. Allele and genotype frequencies were calculated and compared with those of normal controls. The results of our study show a significant decrease in A allele at position UTR 5644 IFN-y, G alleles at position -238 TNF-oc and 166 IL-2, and C allele at position -590 IL-4. TGF- beta haplotype TG/TG increased whereas TGF- beta haplotype CG/CG and IL-10 haplotype GCC/ACC decreased significantly in all patients. Data of this investigation suggest that variations among cytokine gene polymorphisms may contribute to the disease susceptibility. A finding which needs to be fairly clarified in other ethnic groups

Prediction of rejection in renal transplantation by immune parameters

Monitoring of phenotypic characteristics of T-lymphocytes in peripheral blood is commonly performed to give the clinical parameters in the management of kidney transplant recipients. To predict rejection in renal transplantation by immune parameters. 16 non-diabetic kidney transplant candidates [4 females and 12 males, age = 20-65 yr,-first time transplant] were selected. The transplanted patients were divided into two groups based on the rejection during 3 weeks post transplant: group I [n = 9] without rejection and group II [n = 7] with a rejection episode. Immune parameters including lymphocytes subpopulations [by flowcytometry] and immunoglobulin classes [IgM, IgG, IgA and IgE by nephlometric assay] before and 45 days after transplantation were determined. The results of this investigation showed that the level of immunoglobulin IgG, IgM, IgA and IgE decreased post transplantation due to immunosuppressive drugs. CDS, CD4, CDS T cells count, CD56 NK cells count and CD20 B cells count pre- and post-transplantation did not show any significant differences. The amount of IgE [220 vs. 462 Ill/ml], CDS [62% vs. 69.7%] and CD4 [35% vs. 41.3%] cells increased in group II during rejection episode pre-transplantation. In addition, IgA increased pre-transplantation in group I those without rejection episode in comparison with group II with a rejection episode. Forty five days post transplantation IgA [209 vs. 152 mg/dl], IgG [1009 vs. 703 mg/dl] and CD20 [15% vs. 10%] increased in group 1 patients. It is suggestive that pre-transplantation increases IgE, CDS and CD4 are predictive of acute rejection

Th1 and Th2 cytokine gene polymorphism in Iranian patients with chronic myelogenous leukemia

Bakground: lt has been hypothesized that genetic factors other than histocompatibility disparity may play a role in predisposition to developing Chronic Myelogenous Leukemia [CML]. In this regard, Thl and Th2 cytokines and their gene polymorphism seems to be important. Overall expression and secretion of cytokines is dependent, at least in part, on genetic polymorphism [nucleotide variations] within the promoter region or other regulatory sequences of cytokine genes. The majority of polymorphisms described are single nucleotide polymorphism [SNPs]. The objective of this study was to analyze the genetic profile of Thl and Th2 cytokines in 30 Iranian patients with CML and 40 healthy subjects

Methods: In the patients and control subjects, the allelic and genotype frequencies were determined for the cytokine genes. All typing were performed by PCR-SSP assay. Allele and genotype frequencies were calculated and compared with those of normal controls

Results: The results showed that the most frequent alleles in our patients were TGF-3 TG/TG, IL-4 T at position -1089, C at position -590, T at position -33 and IL-10 A at position -1082. Whereas the following alleles - TGF-3 CG/CG and IL-10 C at position -592 - were seen in much lower frequencies

Conclusion: In conclusion, it could be suggested that the frequency of high producing TGF-3 alleles and low producing IL-4 and IL-10 alleles in the CML patients is higher than the normal subjects

Human leukocyte antigen profile in the Zaboli ethnic group living in the South-East Region of Iran

The human leukocyte antigen has become a key component in investigating the genetic relationships between populations. The aim of this study was to determine the genetic diversity of HLA class I and II alleles among Zaboli ethnic group of South-east Iran to establish a database for further investigations on ancestry and the genetic factors contributing to complex diseases in this region. Unrelated individuals from the Southeast geographic location throughout Iran were serologically typed using standard microcytotoxicity assays with commercial and local trays. The ethnic background of each individual was self-defined. HLA profiles were determined in 41 Zaboli populations. The most frequent class I alleles of the Zaboli ethnic group being the following: HLA-A1 [34.1%], -A2 [58.5%], -A11 [29.3%], -A24 [23.9%], -B5 [70.7%], -B16 [26.8%], and -Cw4 [24.4%]. The class II alleles more frequently observed in this group were HLA-DR1 [26.8%], -DR2 [26.8%], -DR3 [31.7%], -DR4 [29.3%], -DR7 [24.4%], -DR8 [22%], -DR11 [48.8%], -DRw52 [73.2%], -DRw53 [53.7%], -DQ1 [53.7%], -DQ2 [31.7%], and -DQ3 [29.3%]. This report utilized a first study of HLA class I and II typed individuals, from widely dispersed areas of Iran. This will help in studies related to disease associations and cadaver organ allocation programmes

Evaluation of Anti- HLA Class I antibodies in chronic rejection of kidney transplantation

Studies have shown that patients who do not produce donor specific and / or panel reactive anti-HLA antibodies have a longer graft survival. The purpose of this study was to evaluate the posttransplant humoral immune response towards HLA-class I antigens and the measurement of the serum creatinine levels which are used in monitoring posttransplant function of kidney. Serum samples from 132 renal transplant recipients were screened for preformed anti-HLA class I panel reactive antibodies [PRA] by means of microlymphocytotoxicity assay. The results revealed the presence of PRA in 26 [19.7%] out of 132 transplanted patients. Graft function was evaluated by measurement of serum creatinine levels which revealed the mean of 1.75 mg/di [SD: 1.08]. Because of clinical significance of presence of different PRA amounts [>10%, > 20% and >50% of panel reactivity] in patients, correlation with kidney function status was analyzed. The obtained data highlighted a higher presence of serum creatinine levels in PRA-positive patients compared to negative patients [P<0.01]. These results [and further studies for class II, ...] can be used to implement new therapeutic strategies to curtail post transplant alloantibodies production and better allografts survival

[Polymorphism in IFN gamma gene has a negative association with type 1 diabetes mellitus in Iranian patients]

Background: type 1A Diabetes Mellitus [T1DM] is a chronic and progressive auto- immune disorder resulting from immune mediated destruction of Langerhans islet beta cells. The etiology of T1DM like the other autoimmune diseases is unknown and many factors are involved, both humoral and cell-mediated immunity have a critical role in T1DM pathogenesis. The cytokines, the immunomodulatory peptides, are responsible for the immune cell recruitment and producing auto-antibodies by the immune effector cells. To evaluate the role of cytokines in sensitivity or resistance to T1DM, we have employed IFN gamma to determine their gene polymorphisms and their association with T1DM

Methods: 30 patient suffering from T1DM and 40 normal control were studied simultaneously .PCR technique was used to characterize the polymorphisms of cytokine. Salting out method was performed for DNA isolation. The polymorphosime of IFN gamma gene was determined on position UTR+5664`5.The PCR products were evaluated by Gel Electerophoresis Technique

Results: there was a significant difference between patient and control group in TT allele IFN gamma gene: p<0.05, RR: 0.39[0.22

Conclusion: in this study, we found a negative association between IFN gamma gene at position 5'UTR+5644 and T1DM in study group that means this allele may be a protective factor against T1DM

Cytokines genes polymorphisms in Iranian patiets with pulmonary tuberculosis

Pulmonary tuberculosis [PTB] has recently become a major problem in developed countries especially in immune compromised HIV infected individuals. Cytokines, their genes and receptors have been implicated in the protective immunity, pathophysiology and development of tuberculosis. In the present study the genotype frequencies of a number of polymorphic genes coding for cytokines or for cytokine receptors have been investigated in a case control study including a group of 40 Iranian PTB patients and 40 healthy individuals. The allelic polymorphism of cytokines SNPs were analyzed according to the protocols of the cytokine component designed for the 13th IHW by the Heidelberg University group. Using PCR-SSP method the following cytokine genes have been determined: IL-1 alpha [T/C -889], IL-1 beta [C/T +3962], IL-1R [C/T pstI 1970], IL-1RA [T/C mspaI 1100], IL-4RA [G/A+1902], IL- 12 [C/A -1188], TGF-beta [C/T codon 10, G/C codon 25], TNF-alpha [G/A -308, G/A -238], IL-2 [T/G -330 G/T +166], IL-4 [T/G -1098, T/C -590, T/C -33], IL-6 [G/C -174, G/A nt 560], IL-10 [G/A -1082, C/T -819, C/A -592]. From IL-1R cluster [pro- inflammatory cytokines] a positive significant association was found at position pstI 1970 C/T polymorphism where the C allele was over presented in the PTB patients [60% vs. 37.5%, P = 0.04].A significant negative association at codon 10 TGF-beta C/T polymorphism has also been shown in our patients, where the T allele was not detected in the patients but 10% of the control subjects expressed this allele [Fisher exact test, P = 0.05]. At this codon allele T [Leucine substitution] is associated with high TGF-beta production. For TNF alpha an insignificant tendency was found at position -308 A/G polymorphism where the G allele carried by 80% of cases and 65% of controls [P = 0.07].At position -238 a negative association was found at the GA polymorphism [10% vs. 25%, P = 0.07]. For IL-6 an insignificant positive association at position -174 C/G polymorphism, G allele [57.5% vs. 37.5, P = 0.07] was found. At the other cytokine genes no specific association were found. In conclusion it is suggested that C allele at position pstI 1970 of IL-1 cluster increases and T allele at codon 10 of TGF-beta decreases in PTB patients

Association of IFN- gamma gene polymorphism with type 1 diabetes in Iranian patients

Type 1 Diabetes [T1D] is a chronic and progressive autoimmune disorder. Cytokines play a critical role in the pathogenesis of T1D. IFN- polymorphism was investigated in T1D and compared with normal controls. Thirty patients suffering fromT1D and 40 normal controls were studied simultaneously using PCR technique. IFN- gene was evaluated at position 5'UTR +5644. There was a significant difference between patient and control groups in TT genotype [P<0.05]. In this study, we found a negative association between IFN- gene at position 5'UTR +5644 and T1D in Iranian patients pointing to T allele as a protective factor against T1D