Editorial 2 2 In recent years, due to increase in infertility rates, Assisted Reproductive technologies (ART) have been very much in use worldwide. The first phase of the study of national infertility amongst the Iranian women, be-tween the ages of 20-40 was completed recently by the Academic Center for Education, Culture and Re-search (ACECR). This study, which was widely reported in the Iranian media, indicates that the infertility rate in this group of women is around 20.2%. The 20.2% infertility rate appears to be above the average reported by the World Health Organization (WHO), which is around 15%. This study was conducted by scientists from Tehran University and Avicenna Research Institute which is also affiliate of the ACECR. This is the first scientific study of the infertility rate in the Iranian women population, and raise awareness of a medical condition that must be addressed by the medical community and health officials in Iran. The use of ART is anticipated to grow rapidly in Iran in the coming years, and this growth will bring about some very important bioethical issues to consider. These include: creation, selection and disposal of unwanted embryos, access, insurance coverage and resource allocation, the rights and responsibilities of individuals who agree to collaborate with each other in donating or receiving reproductive materials. As the use of ART increases in Iran, appropriate ethical laws must be written and passed in the parliament to regulate this rapidly growing industry in order to safeguard the rights of individuals involved in receiving and donating reproductive materials. Also, it would be appropriate for the Iranian Ministry of Health to ap-point a special task force to make sure that all the fertility and infertility centers in Iran abide by the bio-ethical guidelines that are specifically designed and written to safeguard the rights of patients and individuals involved in the use of ART. 54 54 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Editorial 166.pdf ####

Generation of In-vitro Spermatogonial Stem Cells following Genetic Manipulation of Primordial Germ-like Cells 2 2 Research about potential use of stem cells for the development of germ line cells in vitro had been challenged. In the present study, we reported a novel protocol consisting of cocktail growth factor addition for germ cell differentiation followed by transfection. The cells were purificated based on the expression on the cell surface of a protein. This protein is not present in normal cells of mice and does not interfere with cellular function. This cell surface marker is efficiently recognized by monoclonal antibodies. Bone marrow mesenchymal stem cells derived primordial germ like cells were differentiated to spermatogonial stem like cells by inducer cocktail including Retinoic acid (RA)+Leukemia inhibitory factor (LIF)+Basic fibroblast growth factor (bFgF). Co-culture system was used as a feeder under differentiated cells. A 400 bp fragment of spermatogonia-specific Stra-8 locus was enough to direct gene expression to the germ line stem cells. Stra8-CD4HAglo construct was used for purification of premeiotic differentiated cells. Expression of pluripotency (Pou5F1, Nanog, c-Myc) and specific germ cell )Mvh, Piwil2, Stra-8) genes in each stage were analyzed. The purified cells expressed the known molecular markers of PGC-like cells such as Mvh, Piwil2 & Stra-8. The outcomes of qPCR showed that ratio pluripotency of genes expression in selective group significantly decreased (p≤0.05) in the initial differentiation process. This results showed that ratio of Pou5F1, Nanog, c-Myc, Mvh, Piwil2 & Stra-8 expression to purified PGC-like cells were 0.41, 0.204, 1.1, 0.003, 0.184 and 2.276, respectively. Treatment of cells with RA affected up regulation of Stra-8. Although, c-Myc gene as an oncogenic gene had significantly increased (p≤0.05) at the end of differentiation stage compared to initial phase of study, this level of expression could not be tumorgenic. qPCR results of the differentiation stage showed higher expression of Stra-8 in co-culture+cocktail and co-culture groups, Also, there was a significant difference (p≤0.05) in the expression of Pou5F1 & Nanog. Our results suggest that selection and purification of PGC-like cells based on Stra-8 as a pre-meiotic marker is a useful tool for getting in vitro spermatogonial stem cell. This method facilitates identification of safely differentiated germ cells in vitro. 55 63 Zohreh Mazaheri Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University Iran Mansoureh Movahedin Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University Iran Fatemeh Rahbarizadeh Department of Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University Department of Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University Iran Saied Amanpour Vali-e-Asr Reproductive Research Center, Imam Khomeini Hospital, Tehran University of Medical Sciences Vali-e-Asr Reproductive Research Center, Imam Khomeini Hospital, Tehran University of Medical Sciences Iran DifferentiationGene expressionMesenchymal stromal cellsPrimordial germ cellStem cells 82.pdf de Rooij DG. 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Design and Fabrication of Anatomical Bioreactor Systems Containing Alginate Scaffolds for Cartilage Tissue Engineering 2 2 The aim of the present study was to develop a tissue-engineering approach through alginate gel molding to mimic cartilage tissue in a three-dimensional culture system. The perfusion biomimetic bioreactor was designed to mimic natural joint. The shear stresses exerting on the bioreactor chamber were calculated by Computational Fluid Dynamic (CFD). Several alginate/bovine chondrocyte constructs were prepared, and were cultured in the bioreactor. Histochemical and immunohistochemical staining methods for the presence of glycosaminoglycan(GAG), overall matrix production and type II collagen protein were performed, respectively. The dynamic mechanical device applied a linear mechanical displacement of 2 mm to 10 mm. The CFD modeling indicated peak velocity and maximum wall shear stress were 1.706×10-3 m/s and 0.02407 dyne/cm2, respectively. Histochemical and immunohistochemical analysis revealed evidence of cartilage-like tissue with lacunas similar to those of natural cartilage and the production of sulfated GAG of matrix by the chondrons, metachromatic territorial matrix-surrounded cells and accumulation of type II collagen around the cells. The present study indicated that when chondrocytes were seeded in alginate hydrogel and cultured in biomimetic cell culture system, cells survived well and secreted newly synthesized matrix led to improvement of chondrogenesis. 65 74 Anneh Mohammad Gharravi Cellular and Molecular Research Center (CMRC), Faculty of Medicine, Ahvaz Jundishapour University of Medical Sciences (AJUMS) Cellular and Molecular Research Center (CMRC), Faculty of Medicine, Ahvaz Jundishapour University of Medical Sciences (AJUMS) Iran Mahmoud Orazizadeh Cellular and Molecular Research Center (CMRC), Faculty of Medicine, Ahvaz Jundishapour University of Medical Sciences (AJUMS Cellular and Molecular Research Center (CMRC), Faculty of Medicine, Ahvaz Jundishapour University of Medical Sciences (AJUMS Iran Karim Ansari-Asl Electrical Engineering Department, Engineering Faculty, Shahid Chamran University Electrical Engineering Department, Engineering Faculty, Shahid Chamran University Iran Salem Banoni Mechanical Engineering Department, Engineering faculty, Shahid Chamran University Mechanical Engineering Department, Engineering faculty, Shahid Chamran University Iran Sina Izadi Mechanical Engineering Department, Engineering Faculty, Shahid Chamran University Mechanical Engineering Department, Engineering Faculty, Shahid Chamran University Iran Mahmoud Hashemitabar Cellular and Molecular Research Center (CMRC), Faculty of Medicine, Ahvaz Jundishapour University of Medical Sciences (AJUMS) Cellular and Molecular Research Center (CMRC), Faculty of Medicine, Ahvaz Jundishapour University of Medical Sciences (AJUMS) Iran BioreactorsCartilageChondrocyteTissue engineering 83.pdf Ringe J, Kaps C, Burmester GR, Sittinger M. 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Combined Treatment of Androgen-Independent Prostate Cancer Cell Line DU145 with Chemotherapeutic Agents and Lithium Chloride: Effect on Growth Arrest and/or Apoptosis 2 2 Hormone-independent prostate cancer cell lines are resistant to antineoplastic drugs, this study sought to determine the usefulness of lithium chloride as an inhibitor of glycogen synthase kinase-3β to increase the cytotoxic effect of doxorubicin, etoposide or vinblastine antineoplastic drugs on DU145 cells. Combination effect was assessed by using low and IC50 doses of drugs + lithium chloride. Subsequently, cell cycle analysis and p53 levels and its subcellular localization as a key regulator of cell cycle were assessed. Lithium chloride showed cytotoxic effect in a dose and time dependent manner (p<0.001). Both drugs doxorubicin and etoposide in combination with lithium chloride showed higher percent of cells in SubG1 compared to control (p<0.001). Combination of IC50 dose of doxorubicin and lithium chloride led to S phase arrest (p<0.001, compared to control, lithium chloride or doxorubicin alone). Moreover, G2/M arrest was significantly increased when low dose of doxorubicin and vinblastine were combined with lithium chloride (p<0.001, compared to control and lithium chloride alone). DU145 cells were highly sensitive to vinblastine and no significant changes were observed when combined with lithium chloride. The IC50 doses of all three drugs combined with lithium chloride demonstrated decreased cell percent in G1 phase compared to control or lithium chloride alone (p<0.001). Moreover, in the presence of lithium chloride there were increased levels of p53 in cytoplasm and nucleus (p<0.05). Our results suggest that combination of lithium chloride with chemotherapeutic agents may increases their cytotoxic effect on hormone non-responsive human prostate cancer cells. 75 87 Ghamartaj Hossein School of Biology, University College of Science, University of Tehran School of Biology, University College of Science, University of Tehran Iran Vajihe Azimian Zavareh School of Biology, University College of Science, University of TehranDepartment of Regenerative Medicine, Royan Institute for Stem Cell Biology and Technology School of Biology, University College of Science, University of Tehran IranIran Parissa Sahranavard Fard School of Biology, University College of Science, University of Tehran School of Biology, University College of Science, University of Tehran Iran Antineoplastic agentsCombination therapyGSK-3βLithium chloridep53 84.pdf van Brussel JP, van Steenbrugge GJ, Romijn JC, Schröder FH, Mickisch GH. 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Evidence for a role of the TNF receptor-associated death domain protein. J Biol Chem 2001;276(28): 25939-25945. ##Beyaert R, Heyninck K, De Valck D, Boeykens F, van Roy F, Fiers W. Enhancement of tumor necrosis factor cytotoxicity by lithium chloride is associated with increased inositol phosphate accumulation. J Immunol 1993;151(1):291-300.##Munshi A, Pappas G, Honda T, McDonnell TJ, Younes A, Li Y, et al. TRAIL (APO-2L) induces apoptosis in human prostate cancer cells that is inhibitable by Bcl-2. Oncogene 2001;20(29):3757-3765.##Srivastava RK. TRAIL/Apo-2L: Mechanisms and clinical applications in cancer. Neoplasia 2001;3(6):535-546.##Hesry V, Piquet-Pellorce C, Travert M, Donaghy L, Je´gou B, Patard JJ, et al. Sensitivity of prostate cells to TRAIL-induced apoptosis increases with tumor progression: DR5 and caspase 8 are key players. Prostate 2006;66(9):987-995.##Davies MA, Koul D, Dhesi H, Berman R, McDonnell TJ, McConkey D, et al. 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Production of Pentameric Cholera Toxin B Subunit in Escherichia coli 2 2 Cholera toxin B subunit (CTB) has been extensively studied as an immunogen, adjuvant, and inducer of oral tolerance in many investigations. Production of CTB has been carried out in the bacterial, plant, insect and yeast expression systems. In this study the expression of the CTB containing a 6XHis-tagged was performed by Escherichia coli (E.coli) M15. The yield of purified pentameric recombinant CTB was about 1 mg/l. Western blot analysis demonstrated that the recombinant CTB was antigenically active. In addition, GM1-ganglioside ELISA showed that recombinant CTB binds to GM1-gangelioside receptor, confirming disulfide bond formation and proper folding of the recombinant protein in E.coli. Overall, in regard to the vast applications of CTB in medicine, this bacterial expression system will be a fast, cost-effective and simple system for production of pentameric CTB and CTB conjugated proteins. 89 94 Farida Dakterzada Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University Iran Ashraf Mohabati Mobarez Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University Iran Mehryar Habibi Roudkenar Research Center, Iranian Blood Transfusion Organization Research Center, Iranian Blood Transfusion Organization Iran Mehdi Forouzandeh Department of Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University Department of Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University Iran Cholera toxin B SubunitEnzyme-linked immunosorbent assayWestern blotting 85.pdf Wachsmuth IK, Blake PA, Olsvik O, eds. Cholera 2009. Weekly Epidemiol Rec 2010;85:293-308.##Spangler DD. Structure and function of cholera toxin and the related Escherichia coli heat-labile enterotoxin. Microbiol Rev 1992;56(4):622-647.##Jertborn M, Nordstrom I, Kilander A, Czerkinsky C, Holmgren J. Local and systemic immune responses to rectal administration of recombinant cholera toxin B subunit in humans. Infect Immun 2001;69(6):4125-4128.##Bergquist C, Johansson EL, Lagergard T, Holmgren J, Rudin A. Intranasal vaccination of humans with recombinant cholera toxin B subunit induces systemic and local antibody responses in the upper respiratory tract and the vagina. Infect Immun 1997;65(7):2676-2684.##Harakuni T, Sugawa H, Komesu A, Tadano M, Arakawa T. Heteropentameric cholera toxin B subunit chimeric molecules genetically fused to a vaccine antigen induces systemic and mucosal immune responses: a potential new strategy to target recombinant vaccine antigens to mucosal immune systems. Infect Immun 2005;73(9):5654-5665.##Gong Z, Long X, Pan L, Le Y, Liu Q, Wang S, et al. Cloning, expression, purification and characterization of the cholera toxin B subunit and triple glutamic acid decarboxylase epitopes fusion protein in Escherichia coli. Protein Expr Purif 2009;66(2):191-197.##Anjuere F, George-Chandy A, Audant F, Rousseau D, Holmgren J, Czerkinsky C. Transcutaneous immunization with cholera toxin B subunit adjuvant suppresses IgE antibody responses via selective induction of Th1 immune responses. J Immunol 2003;170(3):1586-1592.##Gong Z, Jin Y, Zhang Y. Suppression of diabetes in non-obese diabetic (NOD) mice by oral administration of a cholera toxin B subunit-insulin B chain fusion protein vaccine produced in silkworm. Vaccine 2007;25(8):1444-1451.##Zeighami H, Sattari M, Rezayat M. Cloning and expression of a cholera toxin beta subunit in Escherichia coli. Ann Microbiol 2010;60(3):451-454.##Areas AP, Oliveira ML, Ramos CR, Sbrogio-Almeida ME, Raw I, Ho PL. 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Variation of ATM Gene Expression in Peripheral Blood Cells of Sporadic Breast Carcinomas in Iranian Patients 2 2 The ataxia telangiectasia mutated gene (ATM), candidate for breast cancer susceptibility gene, encode a 350-kDa protein belongs to the core components of DNA-damage response machinery. Female AT carriers have at least 5-fold increase risk for breast cancer. Reduction in ATM expression is shown in multiple studies in breast tissues. We aimed to perform a research to measure the ATM mRNA expression in peripheral blood cells in breast cancer patients. Peripheral blood sample from 40 newly diagnosed, histologically confirmed female breast cancer patients was collected before surgery. Total RNA was isolated from blood cells using the RNX-Plus solution and reverse transcribed into cDNA. Real-time PCR was performed using the 2-∆∆CT method to calculate relative changes in gene expression by REST software. The Relative Quantitation (RQ) mean was 1.27 with the min. and max. equal to 0.20 and 3.34, respectively. Calculation of patient frequencies in different groups revealed that 17.5% had reduced expression lower than two fold decreases and 15% high expression more than two fold increases, but according to REST software there was no up-regulation or down-regulation compared to normal females. The findings of multiple studies consistent with this study indicate that the ATM gene may play an important role in breast cancer development and progression, and ATM expression is down-regulated in breast cancer tissues. Although, some of the results do not support a suppressor role for ATM in the development of sporadic breast cancer, 17.5% of our patients had under expression of ATM mRNA less than two fold relative to control. 95 101 Mohsen Foroughizadeh Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Iran Hossein Mozdarani Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University Iran Keyvan Majidzadeh-A Iranian Center for Breast Cancer, Academic Center for Culture, Education and Research (ACECR) Iran Ahmad Kaviani Faculty of Medicine, Tehran University of Medical Sciences (TUMS) Faculty of Medicine, Tehran University of Medical Sciences (TUMS) Iran Blood cellsBreast neoplasmsmRNAPolymerase chain reactionReal-time 86.pdf Welcsh PL, Schubert EL, King MC. Inherited breast cancer: an emerging picture. Clin Genet 1998;54(6):447-458.##Peto J, Collins N, Barfoot R, Seal S, Warren W, Rahman N, et al. Prevalence of BRCA1 and BRCA2 gene mutations in patients with early-onset breast cancer. J Natl Cancer Inst 1999;91(11):943-949.##Angèle S, Treilleux I, Tanière P, Martel-Planche G, Vuillaume M, Bailly C, et al. Abnormal Expression of the ATM and TP53 Genes in sporadic breast carcinomas. Clin Cancer Res 2000;6(9):3536-3544. ##Lavin M. Role of the ataxia-telangiectasia gene (ATM) in breast cancer. A-T heterozygotes seem to have an increased risk but its size is unknown. BMJ 1998;317(7157):486-487.##Prokopcova J, Kleibl Z, Banwell CM, Pohlreich P. The role of ATM in breast cancer development. Breast Cancer Res Treat 2007;104(2):121-128.##Cuatrecasas M, Santamaria G, Velasco M, Camacho E, Hernandez L, Sanchez M, et al. ATM gene expression is associated with differentiation and angiogenesis in infiltrating breast carcinomas. Histol Histopathol 2006;21(2):149-156.##Feng J, Zou J, Li L, Zhao Y, Liu S. Antisense oligodeoxynucleotides targeting ATM strengthen apoptosis of laryngeal squamous cell carcinoma grown in nude mice. J Exp Clin Cancer Res 2011;30:43.##Swift M, Morrell D, Cromartie E, Chamberlin AR, Skolnick MH, Bishop DT. The incidence and gene frequency of ataxia-telangiectasia in the United States. Am J Hum Genet 1986;39(5):573-583.##Gatti RA, Berkel I, Boder E, Braedt G, Charmley P, Concannon P, et al. Localization of an ataxia-telangiectasia gene to chromosome 11q22–23. Nature 1988;336(6199):577-580.##Werneke U. Ataxia telangiectasia and risk of breast cancer. Lancet 1997;350(9079):739-740.##Swift M. Ionizing radiation, breast cancer, and ataxia-telangiectasia. J Natl Cancer Inst 1994;86(21):1571-1572.##Inskip HM, Kinlen LJ, Taylor AM, Woods CG, Arlett CF. Risk of breast cancer and other cancers in heterozygotes for ataxia-telangiectasia. Br J Cancer 1999;79(7-8):1304-1307.##Clarke RA, Kairouz R, Watters D, Lavin MF, Kearsley JH, Lee CS. Up-regulation of ATM in sclerosing adenosis of the breast. Mol Pathol 1998;51(4):224-226.##Vo QN, Kim WJ, Cvitanovic L, Boudreau DA, Ginzinger DG, Brown KD. The ATM gene is a target for epigenetic silencing in locally advanced breast cancer. Oncogene 2004;23(58):9432-9437.##Wooster R, Ford D, Mangion J, Ponder BA J, Peto J, Easton DF, et al. Absence of linkage to the ataxia telangiectasia locus in familial breast cancer. Hum Genet 1993;92(1):91-94.##Chen JD, Birkholtz GG, Lindblom P, Rubio C, Lindblom. The role of ataxia-telangiectasia heterozygotes in familial breast cancer. Cancer Res 1998;58(7):1376-1379.##Bay JO, Uhrhammer N, Pernin D, Presneau N, Tchirkov A, Vuillaume M, et al. High incidence of cancer in a family segregating a mutation of the ATM gene: possible role of ATM heterozygosity in cancer. Hum Mutat 1999;14(6):485-492.##Rakha EA, Reis-Filho JS, Ellis IO. Combinatorial biomarker expression in breast cancer. Breast Cancer Res Treat 2010;120(2):293-308.##Ha SA, Lee YS, Shin SM, Kim HK, Kim S, Namkoong H, et al. Oncoprotein HCCR-1 expression in breast cancer is well correlated with known breast cancer prognostic factors including the HER2 overexpression, p53 mutation, and ER/PR status. BMC Cancer 2009;9:51.##De Ronde JJ, Hannemann J, Halfwerk H, Mulder L, Straver ME, Vrancken Peeters MJ, et al. Concordance of clinical and molecular breast cancer subtyping in the context of preoperative chemotherapy response. Breast Cancer Res Treat 2010;119(1):119-126.##Tapper W, Hammond V, Gerty S, Ennis S, Simmonds P, Collins A, et al. The influence of genetic variation in 30 selected genes on the clinical characteristics of early onset breast cancer. Breast Cancer Res 2008;10(6):R108.##Gu J, Spitz MR, Zhao H, Lin J, Grossman B, Dinney CP, et al. Roles of tumor suppressor and telomere maintenance genes in cancer and aging-an epidemiological study. Carcinogenesis 2005;26(10):1741-1747.##Jianlin L, Jiliang H, Lifen J, Wei Z, Zhijian C, Shijie C, et al. Variation of ATM protein expression in response to irradiation of lymphocytes in lung cancer patients and controls. Toxicology 2006;224(1-2):138-146.##Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001;25(4):402-408. ##Ye C, Cai Q, Dai Q, Shu XO, Shin A, Gao YT, et al. Expression patterns of the ATM gene in mammary tissues and their associations with breast cancer survival. Cancer 2007;109(9):1729-1735.##Waha A, Sturne C, Kessler A, Koch A, Kreyer E, Fimmers R, et al. Expression of the ATM gene is significantly reduced in sporadic breast carcinomas. Int J Cancer 1998;78(3):306-309.##Kairouz R, Clarke RA, Marr PJ, Watters D, Lavin MF, Kearsley JH, et al. ATM protein synthesis patterns in sporadic breast cancer. Mol Pathol 1999;52(5):252-256.##Kheirollahi M, Mehr-Azin M, Kamalian N, Mehdipour P. Expression of cyclin D2, P53, Rb and ATM cell cycle genes in brain tumors. Med Oncol 2011;28(1):7-14.##Luo L, Lu FM, Hart S, Foroni L, Rabbani H, Hammarstrom L, et al. Ataxia-telangiectasia and T-cell leukemias: no evidence for somatic ATM mutations in sporadic T-ALL or for hypermethylation of the ATM-NPAT/E14 bidirectional promoter in T-PLL. Cancer Res 1998;58(11):2293-2297.##Kovalev S, Mateen A, Zaika AI, O'Hea BJ, Moll UM. Lack of defective expression of the ATM gene in sporadic breast cancer tissues and cell lines. Int J Oncol 2000;16(4):825-831.##Salimi M, Mozdarani H, Majidzadeh K. Expression pattern of ATM and cyclin D1 in ductal carcinoma, normal adjacent and normal breast tissues of Iranian breast cancer patients. Med Oncol 2011 Aug 18. [Epub ahead of print] DOI: 10.1007/s12032-011-0043-5##Tommiska J, Bartkova J, Heinonen M, Hautala L, Kilpivaara O, Eerola H, et al. The DNA damage signalling kinase ATM is aberrantly reduced or lost in BRCA1/BRCA2-deficient and ER/PR/ERBB2-triple-negative breast cancer. Oncogene 2008;27(17):2501-2506.##

Association of HLA-DRB1, DQA1 and DQB1 Alleles and Haplotypes with Common Variable Immunodeficiency in Iranian Patients 2 2 Common Variable Immunodeficiency (CVID) is an antibody deficiency syndrome that often co-occurs in families with selective IgA deficiency (IgAD). This study was designed to investigate the frequency of DR and DQ loci of HLA class II region in common variable immunodeficiency (CVID) patients. Fifteen Iranian patients with CVID or IgAD (mean age 14.6±5.4, range 4-25 years; 9 male and 6 female) and 63 healthy controls were studied. Establishment of B-lymphoblastoid cell lines was performed using Epstein-Barr-virus (EBV) immortalization technique and HLA alleles were typed using polymerase chain reaction based on sequence specific primers (PCR-SSP). DRB1 alleles including DRB1 *04 (p=0.03) and DRB1 *11 (p=0.01) significantly showed higher frequency in the studied subjects. In contrast, DRB1 *301 (p=0.04) and DRB1 *07 (p=0.02) alleles were negatively associated with CVID. For DQB1 and DQA1 loci, DQB1 *0302 (p=0.047) and DQA1 *03011 (p=0.001) demonstrated high frequency in cases, while DQB1 *0201 (p=0.02) and DQA1 *0201 (p=0.01) were detected to be low when compared to controls. Haplotype analysis indicated that frequency of DRB1*04-DQB1*03011-DQA1 *03011 (p=0.02), DRB1 *11-DQB1 *03011-DQA1 *0505 (p=0.047), DRB1 *11-DQA1 *0505 (p=0.04) and DRB1*04-DQA1*03011 (p=0.02) haplotypes were significantly higher in patient group, while only the frequency of the DRB1 *07-DQA1 *0201 haplotype gene was statistically lower in control group (p=0.02). According to the results, it could be deduced that the HLA-DR and DQ loci may contribute to the pathogenesis of CVID or they might be considered as suitable markers for the possibility of the occurrence of this genetic defect. 103 112 Amir Amanzadeh National Cell Bank of Iran, Pasteur Institute of Iran Iran Ali Akbar Amirzargar National Cell Bank of Iran, Pasteur Institute of IranDepartment of Immunology, Faculty of Medicine, Tehran University of Medical Sciences Department of Immunology, Faculty of Medicine, Tehran University of Medical Sciences IranIran Nilufar Mohseni National Cell Bank of Iran, Pasteur Institute of Iran Iran Zohreh Arjang National Cell Bank of Iran, Pasteur Institute of Iran Iran Asghar Aghamohammadi Research Center for Immunodeficiencies, Pediatrics Center of Excellence, Children's Medical Center Research Center for Immunodeficiencies, Pediatrics Center of Excellence, Children's Medical Center Iran Mohammad Ali Shokrgozar National Cell Bank of Iran, Pasteur Institute of Iran Iran Fazel Shokri National Cell Bank of Iran, Pasteur Institute of IranDepartment of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Immunology, School of Public Health, Tehran University of Medical Sciences IranIran HLA antigensAllelesHaplotypesImmunodeficiency 87.pdf Van de Ven AA, Compeer EB, van Montfrans JM, Boes M. B-cell defects in common variable immunodeficiency: BCR signaling, protein clustering and hardwired gene mutations. Crit Rev Immunol 2011;31(2):85-98.##Cunningham-Rundles C, Bodian C. Common variable immunodeficiency: clinical and immunological features of 248 patients. Clin Immunol 1999;92(1):34-48.##Webster ADB. Clinical and immunological spectrum of common variable immunodeficiency (CVID). Iran J Allergy Asthma Immunol 2004;(3)3:103-113.##Hammarstrom L, Vorechovsky I, Webster D. Selective IgA deficiency (SIgAD) and common variable immunodeficiency (CVID). Clin Exp Immunol 2000;120(2):225- 231.##Blancas-Galicia L, Ramírez-Vargas NG, Espinosa-Rosales F. Common variable immunodeficiency. A clinical approach. Rev Invest Clin 2010;62(6):577-582. ##Primary Immunodeficiency Diseases. Report of an IUIS Scientific Committee. Clin Exp Immunol 1999;118(Supplement 1):1-28.##Dhalla F, da Silva SP, Lucas M, Travis S, Chapel H. Review of gastric cancer risk factors in patients with common variable immunodeficiency disorders, resulting in a proposal for a surveillance programme. Clin Exp Immunol 2011;165(1):1-7.##Yel L. Selective IgA deficiency. J Clin Immunol 2010;30(1):10-16.##Kaveri SV, Maddur MS, Hegde P, Lacroix-Desmazes S, Bayry J. Intravenous immunoglobulins in immunodeficiencies: more than mere replacement therapy. Clin Exp Immunol 2011;164 (Suppl 2):2-5.##Borte S, Pan-Hammarström Q, Liu C, Sack U, Borte M, Wagner U, Graf D, Hammarström L. Interleukin-21 restores immunoglobulin production ex vivo in patients with common variable immunodeficiency and selective IgA deficiency. Blood 2009;114(19):4089-4098. ##Moise A, Nedelcu FD, Toader MA, Sora SM, Tica A, Ferastraoaru DE, Constantinescu I. Primary immunodeficiencies of the B lymphocyte. J Med Life 2010;3(1):60-63. ##De Gast GC, Wilkins SR, Webster AD, Rickinson A, Platts-Mills TA. Functional immaturity of isolated B cells from patients with hypogammaglobulinaemia. Clin Exp Immunol 1980; 42(3):535-544.##Hammarstrom L, Smith CIE. Genetic approach to common variable immunodeficiency and IgA deficiency. In: Ochs H, Smith CI, Puck J (eds). Primary immunodeficiency diseases a molecular and genetic approach. Oxford: Oxford University Press;1999,250-262.##Schaffer FM, Palermos J, Zhu ZB, Barger BO, Cooper MD, Volanakis JE. Individuals with IgA deficiency and common variable immunodeficiency share polymorphisms of major histocompatibility complex class III genes. Proc Natl Acad Sci USA 1989; 86(20):8015-8019.##Olerup O, Smith CIE, Björkander J, Hammarström L. Shared HLA class II-associated genetic susceptibility and resistance, related to the HLA-DQB1 gene, in IgA deficiency and common variable immunodeficiency. Proc Natl Acad Sci USA 1992; 89(22):10653-10657.##Carvalho Neves Forte W, Ferreira De Carvalho Jr F, Damaceno N, Vidal Perez F, Gonzales Lopes C, Mastroti RA. Evolution of IgA deficiency to IgG subclass deficiency and common variable immunodeficiency. Allergol Immunopathol 2000; 28(1):18-20.##Español T, Catala M, Hernandez M, Caragol I, Bertran JM. Development of a common variable immunodeficiency in IgA deficient patients. Clin Immunol Immunopathol 1996;80(3 Pt 1):333-335.##Ishizaka A, Nakanishi M, Yamada S, Sakiyama Y, Matsumoto S. Development of hypogammaglobulinaemia in a patient with common variable immunodeficiency. Eur J Pediatr 1989;149(3):175–176.##Koistinen J. Familial clustering of selective IgA deficiency. Vox Sang 1976; 30(3):181-190.##Oen K, Petty RE, Schroeder ML. Immunoglobulin A deficiency: genetic studies. Tissue Antigens 1982;19(3):174-182.##Conley ME, Cooper MD. Immature IgA B cells in IgA deficient patients. N Engl J Med 1981;305(9):495-497.##Cooper MD, Lawton AR, Bockman DE. Agammaglobulinaemia with B lymphocytes. Specific defect of plasma-cell differentiation. Lancet 1971;2(7728):791-794.##Eisenstein EM, Chua K, Strober W. B cell differentiation defects in common variable immunodeficiency are ameliorated after stimulation with anti-CD40 antibody and IL- 10. J Immunol 1994;152(12):5957-5968.##Farrington M, Grosmaire LS, Nonoyama S, Fischer SH, Hollenbaugh D, Ledbetter JA, et al. CD40 ligand expression is defective in a subset of patients with common variable immunodeficiency. Proc Natl Acad Sci USA 1994; 91(3):1099-1103.##De La Concha EG, Fernandez-Arquero M, Martinez A, Vidal F, Vigil P, Conejero L, et al. HLA class II homozygosity confers susceptibility to common variable immunodeficiency (CVID). Clin Exp Immunol 1999;116(3):516-520.##Olerup O, Smith CIE, Hammarström L. Different amino acids at position 57 of the HLA-DQβ chain associated with susceptibility and resistance to IgA deficiency. Nature 1990;347(6290):289-290.##Schroeder HW Jr, Zhu ZB, March RE, Campbell RD, Berney SM, Nedospasov SA, et al. Susceptibility locus for IgA deficiency and common variable immunodeficiency in the HLA-DR3, -B8,-A1 haplotypes. Mol Med 1998;4(2):72-86. ##Shokri F, Mageed RA, Maziak BR, Jefferis R. Expression of VHIII-associated cross-reactive idiotype on human B lymphocytes. Association with staphylococcal protein A binding and Staphylococcus aureus Cowan I stimulation. J Immunol 1991;146(3):936-940.##Olerup O, Zetterquist H. HLA-DR typing by PCR amplification with sequence-specific primers (PCR-SSP) in 2 hours: an alternative to serological DR typing in clinical practice including donor-recipient matching in cadaveric transplantation. Tissue Antigens 1992;39(5):225-235. ##Wollheim FA, Belfrage S, Coster C, Lindholm H. Primary "acquired" hypogammaglobulinemia; clinical and genetic aspects of nine cases. Acta Med Scand 1964;176:1-16.##Fiore M, Pera C, Delfino L, Scotese I, Ferrara GB, Pignata C. DNA typing of DQ and DR alleles in IgA-deficient subjects. 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