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Background: According to the WHO's recommendation for developing countries, Bacillus Calmette-Guerin (BCG) vaccination has been implemented in some countries as part of national vaccination programs at birth. Although it is generally considered safe, some complications may occur; including BCGitis (local) or BCGosis (systemic), ranging from mild like local abscesses to fatal impediments like osteomyelitis and disseminated BCG infection. This study aimed to determine the spectrum of inborn errors of immunity (IEI) in BCG-vaccinated neonates experiencing local or systemic complications. Methods: In this cross-sectional study, we investigated Iranian children referred to the Immunology, Asthma, and Allergy Research Institute (IAARI) between 2007-2023 for suspected immunodeficiency. Medical history was recorded, and primary screening tests for immunodeficiency were conducted for all cases. For suspected cases, more advanced immunologic investigations were performed to reach a definitive diagnosis. Furthermore, the study incorporated the documented genetic findings of the patients under investigation. All patients with inborn error of immunity who had a history of BCG vaccine complications within the first year of vaccination were enrolled in the study. Results: We investigated 3,275 cases suspected of IEI, identifying197 patients with both IEI and BCG vaccine complications. Among these, 127 (64.5%) were male. Symptoms began at or before 3 months of age in 64.8% of the cases, and parental consanguinity was reported in 79.2%. Genetic diagnoses were confirmed in 108 patients. Of the 197 patients, 108 (54.8%) had BCGitis, while 89 (45.2%) experienced systemic complications (BCGosis). A family history of IEI, BCG-related complications, and unexplained deaths were observed in 20.3%, 12.2%, and 29.9% of cases, respectively. Furthermore, 46.2% had at least one of these three risk factors in their history. Conclusions: Early BCG vaccine complications may indicate an underlying immunodeficiency, particularly when there is a positive family history of BCG complications, immunodeficiency, or unexplained deaths. Nation-wide vaccination protocols should address this issue by delaying inoculation to allow for immunological screening of suspected immunodeficient patients, thereby preventing BCG vaccine-related morbidity and mortality. [ABSTRACT FROM AUTHOR]

Background: Primary immunodeficiency diseases (inborn errors of immunity) with partial albinism are a group of autosomal recessive syndromes including Chediak Higashi Syndrome (CHS), Griscelli Syndrome type 2 (GS2), Hermansky‐Pudlak Syndromes type 2 and 10 (HPS2, HPS10), Vici syndrome and P14/LAMTOR2 deficiency. Methods: Twenty‐five patients including 10 CHS, 10 GS2, and 5 HPS2 were evaluated in this study within the last 10 years. Five cases with oculocutaneous albinism (OCA) and 5 healthy subjects without albinism were used as two control groups. Genetic analyses were performed by whole exome or panel sequencing or targeted Sanger sequencing. Subsequently, leukocyte granules in peripheral blood smear and hair shaft were examined as screening tests. Results: Giant granules were only presented in the leukocytes cytoplasm of 10/10 CHS patients. The uneven cluster of pigments and giant melanin granules in hair samples were observed in 10/10 GS2 and 10/10 CHS patients, respectively. In both 5/5 OCA and 5/5 HPS2 patients, there were regular pigments in the middle of hair shafts. Genetic analyses were performed for all patients, revealing 7 novel variants in LYST gene for CHS patients and 4 novel variants in AP3B1 for HPS2 patients. Conclusion: Receiving hematopoietic stem cell transplantation (HSCT) in a timely manner is crucial in CHS and GS2 patients; therefore, screening tests may provide a vital clue for early diagnosis in these patients. However, the final confirmation of CHS, GS2, and HPS2 disorders is done by genetic assay. [ABSTRACT FROM AUTHOR]

Background: In order to support the comprehensive classification of Leukocyte Adhesion Deficiency‐I (LAD‐I) severity by simultaneous screening of CD11a/CD18, this study assessed clinical, laboratory, and genetic findings along with outcomes of 69 LAD‐I patients during the last 15 years. Methods: Sixty‐nine patients (40 females and 29 males) with a clinical phenotype suspected of LAD‐I were referred to Immunology, Asthma, and Allergy research institute, Tehran, Iran between 2007 and 2022 for further advanced immunological screening and genetic evaluations as well as treatment, were enrolled in this study. Results: The diagnosis median age of the patients was 6 months. Delayed umbilical cord separation was found in 25 patients (36.2%). The median diagnostic delay time was 4 months (min–max: 0–82 months). Forty‐six patients (66.7%) were categorized as severe (CD18 and/or CD11a: below 2%); while 23 children (33.3%) were in moderate category (CD18 and/or CD11a: 2%–30%). During the follow‐ups, 55.1% of children were alive with a mortality rate of 44.9%. Skin ulcers (75.4%), omphalitis (65.2%), and gingivitis (37.7%) were the most frequent complaints. Genetic analysis of the patients revealed 14 previously reported and three novel pathogenic mutations in the ITGB2 gene. The overall survival of patients with and without hematopoietic stem cell transplantation was 79.3% and 55.6%, respectively. Conclusion: Physicians' awareness of LAD‐I considering delayed separation of umbilical cord marked neutrophilic leukocytosis, and variability in CD11 and CD18 expression levels, and genetic analysis leads to early diagnosis and defining disease severity. Moreover, the prenatal diagnosis would benefit families with a history of LAD‐I. [ABSTRACT FROM AUTHOR]

Griscelli syndrome type 2 (GS2) is an autosomal recessive immunodeficiency characterized by hair hypopigmentation, recurrent fever, hepatosplenomegaly and pancytopenia. This study aims to find new genetic changes and clinical features in 18 children with GS2 caused by the RAB27A gene defect. In all, 18 Iranian children with GS2 who presented with silver grey hair and frequent pyogenic infection were included in this study. After recording demographic and clinical data, PCR sequencing of the RAB27A gene was performed for all exons and exon–intron boundaries. Two patients in this study were subjected to whole‐exome sequencing followed by Sanger sequencing. Light microscopy study of hair showed large irregular clumps of pigment with the absence of giant granules on the blood smear. Mutation analysis of the RAB27A gene identified two novel missense mutations as homozygous in a patient, one in exon 2, c.140G>C and another in exon 4, c.328G>T. In addition, for 17 other patients, 6 reported mutations were obtained including c.514_518delCAAGC, c.150_151delAGinsC, c.400_401delAA, c.340delA, c.428T>C and c.221A>G. The mutation c.514_518delCAAGC was the most frequent and found in 10 patients; this mutation may be considered a hotspot in Iran. Early diagnosis and treatment of RAB27A deficiency can contribute to better disease outcomes. In affected families, genetic results could be urgently needed to make a timely decision about haematopoietic stem cell transplantation and prenatal diagnosis. [ABSTRACT FROM AUTHOR]

Chronic granulomatous disease (CGD) is a rare primary immunodeficiency disorder more common in autosomal recessive (AR) than X-linked in Iran. This study aimed to assess whether having a child with AR-CGD would increase the likelihood of the next child being affected by CGD. Ninety-one families with at least one child affected by AR-CGD entered this study. Out of the 270 children, 128 were affected by AR-CGD. We used a cross tab for the odds ratio (OR) calculation, in which exposure to a previously affected child and the next child’s status were evaluated. This study illustrated that the chances of having another child afflicted with AR-CGD are significantly increased if the previous child had AR-CGD (OR=2.77, 95% CI=1.35-5.69). Although AR disorders affect 25% of each pregnancy, we showed that the chance that the next child would be affected by CGD, given that the previous child was affected, is 2.77 times greater than in families with a normal child. It is recommended to warn families with one or more affected children to evaluate the risk of CGD in their subsequent pregnancies with prenatal diagnosis. [ABSTRACT FROM AUTHOR]

Chronic granulomatous disease (CGD) is a life-threatening immunodeficiency condition. To date, hematopoietic stem cell transplantation (HSCT) is the only curative modality. We prospectively studied the outcomes of fifteen CGD patients undergoing HSCT with fludarabine and melphalan plus anti-thymocyte globulin (ATG). Most of the donors were fully matched siblings (n = 12). Cyclosporine A and methylprednisolone were used for graft-versus-host disease (GVHD) prophylaxis. CGD diagnosis had been suspected upon clinical symptoms and was confirmed in all patients by an abnormal neutrophil functional assay. The three-year overall survival (OS) and event-free survival (EFS) rates were 73.3% and 46.7%, respectively. With the median follow-up time of 33.12 months, the mean OS and EFS were 42.6 and 26.8 months; respectively. Eleven patients (73.33%) achieved full donor chimerism. Two stable mixed chimerisms with no sign of the underlying disease (13.33%) and two secondary graft failure (13.33%) occurred as well. The cumulative incidence of transplant-related mortality was 23.1% and it was two times more in adults compared with children. Three years GVHD-FS (free survival) was 57.8% in all patients and it was 70% and 42.9% in children and adults, respectively. Our results indicate that fludarabine, melphalan, and ATG have relatively favorable outcomes in CGD patients. Also, we suggest that HSCT should be performed as soon as a suitably matched donor is found. [ABSTRACT FROM AUTHOR]

T-cell receptor excision circles (TREC)/Kappa-deleting recombination excision circles (KREC) assay has been recently recognized for detecting patients with primary (T- and/or B-cell) immunodeficiency (PID). We aimed to investigate the alterations of these biomarkers in some combined immunodeficiency patients compared to the healthy controls in different age groups. TREC and KREC were assessed in a total of 82 PID patients, most of them with exact genetic diagnosis (3 months to 42 years); using quantitative real-time-polymerase chain reaction (PCR). Patients had a final diagnosis of common variable immunodeficiency (n=23), ataxiatelangiectasia (AT) (n=17), hyper-IgE syndrome (HIES) (7 with DOCK8 deficiency, 4 with signal transducer and activator of transcription 3 (STAT3) deficiency, and 8 children with unknown genetic defects), Wiskott-Aldrich syndrome (WAS) (n=20), purine nucleoside phosphorylase (PNP)deficiency(n=1), dedicator of cytokinesis2 (DOCK2) deficiency (n=1), recombinase activating gene1 (RAG1) deficiency (n=1). Very low to zero amounts of TREC and/or KREC were detected in 14 out of 23 cases of common variable immunodeficiency (CVID), 14 out of 17 cases of AT, 8 out of 20 cases of WAS, 6 out of 7 cases of DOCK8-deficiency patients, 4 out of 8 cases of HIES with unknown genetic defects and all patients with defects in DOCK2, PNP, and RAG1. STAT3-deficient patients were normal for both biomarkers. All patients showed a significant difference in both markers compared to age-matched healthy controls. Our findings highlight that apart from severe types of T/B cell defects, this assay can also be used for early diagnosis the patients with late-onset of disease and even PIDs without a positive family history. [ABSTRACT FROM AUTHOR]

This study aims to investigate the role of prenatal diagnosis (PND) in Iranian couples with a previous history of primary immunodeficiency disorders (PIDD) in their family. All referred couples with a family history of PIDD and a tendency for PND were included in this project. Based on gestational age, chorionic villus sampling (CVS) was performed to analyze the molecular defect of the fetus according to the previous gene defect of the affected case in the family. Postnatal confirmation was performed by immunological screening tests. In a total of 100 cases, CVS was not evaluated in 19 patients due to unwillingness (n=5), late prenatal referral (n=7), miscarriage before CVS (n=3), and female fetus with x-linked diseases in previous children (n=4). In the remaining 81 patients, heterozygous and homozygous mutations were found in 33 and 23 cases, respectively. The hemizygous mutation was obtained in 6 and no pathogenic mutations were found in 19 individuals. Postnatal evaluations revealed that a total of 65 babies were healthy, 32 fetuses were aborted (3 cases before CVS, 2 spontaneous abortions of a healthy and as affected fetus in the CVS subgroup, and 27 cases were aborted due to therapeutic causes). One fetus from the heterozygous subgroup was spontaneously aborted with severe combined immunodeficiency (SCID) and one fetus from the homozygous subgroup that was supposed to be healthy was affected by the autosomal dominant-chronic granulomatous disease (AR-CGD). The diagnostic error was 1.2%. PND is highly recommended in families with a history of PID in their previous child to prevent an affected baby being born and to reduce the government, family, and personal burden of these diseases. [ABSTRACT FROM AUTHOR]

Complete TYK2 deficiency (IMMUNODEFICIENCY 35 OMIM (611521)) is a rare disorder, inherited as an autosomal recessive (AR) trait, which has been previously described in nine patients from seven unrelated kindreds [[1]-[3]]. Following mandatory BCG vaccination at birth in Iran and some other countries, patients with certain inborn errors of immunity may develop BCG disease that usually appears 3 months after the vaccination [[9]-[12]]. In line with other patients with complete AR TYK2 deficiency, our patient also suffers from viral disease, particularly with herpes viruses, attributable to an impaired IFN- -mediated immunity (2, 6). Importantly, both our patient and the patient reported in 2016 by Fuchs et al. carry the same TYK2 mutation and we thus infer that our patient would show a lack of TYK2 expression despite never tested. [Extracted from the article]

Familial haemophagocytic lymphohistiocytosis (FHL) is a rare disorder of immune dysregulation. FHL inherited in an autosomal recessive pattern is classified into five subtypes based on underlying genetic defects. Mutations in four genes including PRF1, UNC13D, STX11 and STXBP2 are responsible for FHL2 to FHL5 respectively. The cause of FHL1 is associated with mutations in an unknown gene located at 9q21.3-22. This study aims to report the clinical features and genetic results of nine Iranian patients suffering from - haemophagocytic lymphohistiocytosis. Nine patients (five males and four females) suspected to FHL whose genetic evaluation of PRF1 and STX11 revealed no mutations, were entered the study to investigate UNC13D mutations. Primers were designed to amplify all coding regions and exon-intron boundaries of the gene. PCR products were then sequenced and analyzed by sequence analysis tools including BLAST. The most frequent clinical manifestations observed in the patients were fever and hepatosplenomegaly. In this study, five mutations were detected in UNC13D including four novel mutations (c.1434_1446delACCCATGGTGCAGinsTGGTGCT, c.1933C>T, c.1389+1G>C and c.2091+1G>A) besides to a previously reported deletion (c.627delT). The pathogenicity of the missense mutation was assessed using online prediction tools including SIFT and PolyPhen2. The study results may provide valuable information for genetic counseling especially for those who have a history of immunodeficiency diseases in their family and can be used for prenatal diagnosis. [ABSTRACT FROM AUTHOR]

Chronic granulomatous disease (CGD) is a rare genetic disorder of neutrophil activity, resulting in increased rate of recurrent infections with catalase-positive bacteria and fungi, as well as various autoimmune diseases such as sarcoidosis, rheumatoid arthritis, and discoid and/or systemic lupus erythematosus. Few reports have reported lupus erythematosus (LE) in patients with X-linked CGD (XL-CGD) and carriers, and very few in autosomal recessive CGD (AR-CGD). Here, we present 5 patients with CGD developing LE at different ages to emphasize on the importance of appropriate follow-up and treatment in patients with CGD with clinical signs and symptoms of autoimmune diseases and even in those with negative serologic results. [ABSTRACT FROM AUTHOR]

In this study, we first tried to determine whether the expression level of 9 miRNAs in the peripheral blood CD8+ T cells of asthmatic patients varies from that of controls, and secondly, we investigated the effects of fluticasone furoate and vilanterol on the expression level of these miRNAs. Fifteen subjects including 8 healthy individuals and 7 asthmatic patients were included in this study. CD8+T cells were isolated from participants' peripheral blood by a negative selection method using magnetic-activated cell sorting (MACS). The expression of 9 miRNAs was examined between the healthy individuals and asthmatic patients. Then the expression level of 9 miRNAs before and after treatment with the drugs was examined by quantitative real-time PCR. No significant changes in the expression level of 9 miRNAs were observed in asthmatic patients compared to the healthy controls. Fluticasone and vilanterol, in combination, had the greatest effect on miRNA expression. MiR-150 and miR-106a were the most and the least miRNAs, respectively, present in CD8+ T cells of patients and controls. MiR-106a and miR-126 had a positive correlation in CD8+ cells of asthmatic patients. Although no significant difference in the expression level of studies miRNAs was observed, the correlations among miRNAs were significant. Therefore, we suggest that the correlation between miRNAs would be a very important factor in physiological and pathological conditions in healthy individuals and asthmatic patients. Such a miRNA-miRNA correlation network can be even more critical than any changes in the variation of their expression in the CD8+ T cells. [ABSTRACT FROM AUTHOR]

Purpose: Mendelian susceptibility to mycobacterial disease (MSMD) is a rare primary immunodeficiency, triggered by non-tuberculous mycobacteria or Bacillus Calmette-Guérin (BCG) vaccines and characterized by severe diseases. All known genetic etiologies are inborn errors of IFN-γ-mediated immunity. Here, we report the molecular, cellular, and clinical features of patients from 15 Iranian families with disseminated disease without vaccination (2 patients) or following live BCG vaccination (14 patients). Methods: We used whole blood samples from 16 patients and 12 age-matched healthy controls. To measure IL-12 and IFN-γ, samples were activated by BCG plus recombinant human IFN-γ or recombinant human IL-12. Immunological assessments and genetic analysis were also done for the patients. Results: Eight patients affected as a result of parental first-cousin marriages. Seven patients originated from multiplex kindred with positive history of death because of tuberculosis or finding the MSMD-related gene mutations. Two patients died due to mycobacterial disease at the ages of 8 months and 3.7 years. The remaining patients were alive at the last follow-up and were aged between 2 and 13 years. Patients suffered from infections including chronic mucocutaneous candidiasis (n = 10), salmonellosis (n = 2), and Leishmania (responsible for visceral form) (n = 2). Thirteen patients presented with autosomal recessive (AR) IL-12Rβ1 deficiency, meaning their cells produced low levels of IFN-γ. Bi-allelic IL12RB1 mutations were detected in nine of patients. Three patients with AR IL-12p40 deficiency (bi-allelic IL12B mutations) produced low levels of both IL-12 and IFN-γ. Overall, we found five mutations in the IL12RB1 gene and three mutations in the IL12B gene. Except one mutation in exon 5 (c.510C>A) of IL12B, all others were previously reported to be loss-of-function mutations. Conclusions: We found low levels of IFN-γ production and failure to respond to IL12 in 13 Iranian MSMD patients. Due to complicated clinical manifestations in affected children, early cellular and molecular diagnostics is crucial in susceptible patients. [ABSTRACT FROM AUTHOR]

T-cell receptor excision circles (TRECs) and κ -deleting recombination excision circles (KRECs) are recently used for detection of T or B cell lymphopenia in neonates based on region -specific cutoff levels. Here, we report cutoffs for TREC and KREC copies useful for newborn screening and/or diagnosis of primary immunodeficiency diseases (PID) in Iran. DNA was extracted from a single 3.2 mm punch of dried blood spots collected from 2160 anonymized newborns referred to two major referral health centres between 2014 and 2016. For refinement of the cutoffs, 51 patients with a definite diagnosis of severe combined immunodeficiency, X -linked agammaglobulinaemia and combined immunodeficiency, including ataxia telangiectasia, human phosphoglucomutase 3 and Janus kinase -3 deficiency, as well as 47 healthy controls were included. Samples from patients with an X -linked hyper -IgM -syndrome, Wiskott -Aldrich syndrome and DNA ligase 4 deficiency were considered as disease controls. Triplex -quantitative real -time PCR was used. Cutoffs were calculated as TRECs < 11 and KRECs < 6 copies with an ACTB > 700 copies with sensitivity of 100% for TREC and 97% for KREC. Among thirty anonymized newborn samples (1.5%) with abnormal results for TREC and/or KREC, only twenty -one available cases were retested and shown to be in the normal range except for three samples (0.15%). All of the patients with a definitive diagnosis were correctly identified based on our established TREC/KREC copy numbers. Determining cutoffs for TREC/ KREC is essential for correctly identifying children with PID in newborn screening. Early diagnosis of PID patients enables appropriate measures and therapies like stem cell transplantation. [ABSTRACT FROM AUTHOR]

Chronic granulomatous disease (CGD) is a rare primary immunodeficiency disorder due to a genetic defect in one of the components of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex. This complex is composed of membrane-bound gp91-phox and p22-phox subunits, and cytosolic subunits consisting of p47-phox, p67-phox, and p40-phox. A mutation in CYBB gene encoding gp91-phox located on chromosome Xp21.1, leads to X-linked CGD. Herein, we report a 4-year-old Iranian boy presented with episodes of recurrent fever, cervical lymphadenopathy, and abdominal abscesses. Mutation analysis of the CYBB gene in the patient indicated a one-nucleotide deletion, c.316delT, resulting in p.W106GfsX. [ABSTRACT FROM AUTHOR]

Chronic granulomatous disease (CGD), a rare inherited primary immunodeficiency disorder, is caused by mutation in any one of the genes encoding components of nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase enzyme. NCF2 gene (encoding P67-phox component) is one of them and its mutation is less common to cause CGD (around 5-6%). Here, we assessed mutation analysis of NCF2 in 4 CGD patients with p67-phox defect in Iran. These patients showed classical CGD symptoms. NCF2 sequence analyses revealed two different homozygous mutations including a nonsense mutation in exon 4, c.304C>T (Arg 102X) in one case and a CA deletion in exon 13 (Leu346fsX380) in one brother and sister; the latter is a new mutation which has not been reported in previous studies. In another patient in whom the attempts to amplify exon 2 individually from genomic DNA were unsuccessful, PCR amplification of exon 2 revealed no band of this exon on agarose gel. A PCR amplification mix of exon 2 and exon 7, with an internal control, confirmed the lack of exon 2 in this patient. Although a gross deletion in other exons of NCF2 has been previously reported, a large deletion encompassing exon 2 has been not reported yet. This abstract was also presented in ESID 2012, Florence, Italy. [ABSTRACT FROM AUTHOR]

Background: Chronic granulomatous disease (CGD) is a rare immunodeficiency due to a genetic defect in one of the NADPH-oxidase components. We studied CGD inheritance forms (autosomal recessive (AR) or X-linked (XL)) and AR-CGD subtypes in Iran. Methods: Clinical and functional investigations were conducted in 93 Iranian CGD patients from 75 families. Results: Most of the patients were AR-CGD (87.1%). This was related to consanguineous marriages ( p = 0.001). The age of onset of symptoms and diagnosis were lower in XL-CGD compared with AR-CGD ( p < 0.0001 for both). Among AR-CGD patients, p47 phox defect was the predominant subtype (55.5%). The most common clinical features in patients were lymphadenopathy (65.6%) and pulmonary involvement (57%). XL-CGD patients were affected more frequently with severe infectious manifestations. Conclusions: Although XL-CGD is the most common type of the disease worldwide, only 12 patients (12.9%) were XL-CGD in our study. The relatively high frequency of AR-CGD is probable due to widely common consanguineous marriages in Iran. [ABSTRACT FROM AUTHOR]

One of the rarest forms of chronic granulomatous disease (CGD) is caused by mutations in CYBA, which encodes the p22-phox subunit of the phagocyte NADPH oxidase, leading to defective intracellular killing. This study investigated eight patients (six males and two females) from seven consanguineous, unrelated families with clinical CGD, positive family history and p22-phox deficiency. Mutation analysis of CYBA showed six different novel mutations: deletion of exons 3, 4 and 5; a missense mutation in exon 6 (c.373G>A); a splice site mutation in intron 5 (c.369+1G>A); a frameshift in exon 6 (c.385delGAGC); a frameshift in exon 3 (c.174delG); and a frameshift in exon 4 (c.223delC). [ABSTRACT FROM AUTHOR]

Objective: Hemophagocytic lymphohistiocytosis (HLH) is a rare immunodeficiency disorder recognized by high fever without response to antibiotics, hepatosplenomegaly, cytopenia and hemophagocytosis. HLH can be classified into two major forms: primary and secondary. Familial hemophagocytic lymphohistiocytosis (FHL), a type of primary HLH in which mutations in PRF1 UNC13D, STX11, STXBP2 genes cause the disease. Since genetic studies in STX11 and PRF1 have been investigated in Iranian FHL patients and no mutations found in some of them, this study aims to include UNC13D gene to FHL diagnosis panel to help patients diagnose early and accurate and do prenatal and carrier status. Material and Methods: Patients with HLH diagnosis and without mutation in PRF1 and STX11 genes entered this study. Primers were designed for exon and exonintron regions of UNC13D gene. After genomic DNA was extracted from whole blood samples, PCR technique was done and followed by direct sequencing of the products. Finally, sequencing results were analyzed. Results: Mutation analysis of UNC13D gene revealed several mutations including one novel splicing mutation (c.2091+1G>A), one missense mutation (c.1933C>T), one frame shift deletion (c.627Tdel) and three single nucleotide polymorphisms (SNPs) (c.888G>C, c.2601A>G, c.3183A>G). Conclusion: Considering the mutations detected in UNC13D gene and its association with FHL, elucidation of genetic study of this gene is necessary for accurate diagnosis and appropriate treatment and can help us extend our knowledge of this heterogeneous disease. [ABSTRACT FROM AUTHOR]

Objective: Biobank is a collection of resources containing biological and genetic materials that help store valuable samples. Establishing a biobank makes easy to access samples for assessing their characteristics. Our biobank consists of four separate biobanks including DNA, HLA, serum, and cell banks. Our aim is to standardize banking the samples of patients and their families who referred to Immunology, Asthma and Allergy Research Institute (IAARI) and suffered from primary immunodeficiency (PID), asthma and allergy. Materials and Methods: IAARI's biobank was established in 2006 with DNA and serum banks, then cell and HLA banks were added in 2012. All of the cases with diagnosis of PID, asthma and allergy entered in this registry and after taking informed consent, their samples were stored in the biobank. Results: Up to now, we stored a total of 3288 DNA samples from PID patients and their relatives, 450 serums from allergic patients, 186 samples of asthmatic patients, more than 700 HLA samples of healthy donors' volunteers, and 43 cell samples from PID patients. Conclusion: This biobank can be used to early diagnosis of PID genetics, and finding matched hematopoietic stem cell transplantation (HSCT) candidates for them. In addition, it can be helpful for investigating prospective genetic and familial surveys. This also decreases need for cost and resampling from patients. [ABSTRACT FROM AUTHOR]

Objective: Leukocyte adhesion deficiency (LAD) is one of the rare immunodeficiency diseases clinically characterized by recurrent severe bacterial infections, periodontitis, delayed umbilical cord separation, absence of pus formation, periodontitis, delayed wound healing, and often persistent leukocytosis. Three types of the disease were discussed. With autosomal recessive inheritance and a prevalence of 1 in 1,000,000 live births, Type I (LADI) is caused by mutations in the gene ITGB2, located on chromosome 21 (21q22.3), encoding for CD18 protein. More than 110 distinct mutations in ITGB2 gene have been identified and registered at Human Gene Mutation Database (HGMD; http://www.hgmd.cf.ac.uk/ac/all.php).This study aims to find the disease causing mutations in ITGB2 gene for helping future life in patient families via genetic counseling and prenatal diagnosis. Material and Methods: All of the patients with LAD1 diagnosis in the base of Flow cytometry (defect in CD18 and CD11 a, b, c) who were referred to IARRI during 2 years entered the study. 2 mL EDTA blood was obtained from all patients and their parents and genomic DNA was extracted. Polymerase chain reaction (PCR) for encoding exons and exon-intron boundaries of ITGB2 gene was performed. PCR results were direct sequenced in both orientations. Results: Sequencing amplified regions by PCR revealed a total of five different new mutations in five exons. These included three missense mutations, (c.382G>A in exon 6, c.715G>A in exon 7 and c.850G>A in exon 8) and two deletions (c.1907delA in exon 15 and c.1590_1594delCGGGC in exon 13). Mutation c.1907delA was found in two unrelated families. The last mutation was not previously reported. In fact, four out of 5 mutations have been reported previously. However, we find the mutation c.1590_1594delCGGGC neither in HGMD nor in other human mutation databases. Deletion of 5 nucleotides in this mutation results in a frame shift in open reading frame of CD18 protein and causes a premature stop codon at that position, p.Tyr530*. In all cases, mutation found in the patients was observed in their parents. Conclusion: The found mutations in this study can be used for genetic counseling and prenatal diagnosis of LAD I. [ABSTRACT FROM AUTHOR]

Objective: Primary immunodeficiency disorders (PID) are a heterogeneous group of genetic disorders resulting from quantitative or functional defects in immune cells. Recently, T-cell receptor excision circles (TRECs) and κ-deleting recombination excision circles (KRECs) have been used for the detection of T and B cell lymphopenia in newborn screening programs based on region-specific cutoff levels. Here, we report the newborn screening program for obtaining the cutoff levels for TREC and KREC copy numbers that may be applied in our newborn screening program and as a diagnostic tool in infants with a family history of PID in Iran. Materials and methods: DNA was extracted from a single 3.2mm punch from dried blood spots collected from 2160 anonymized newborns, which were obtained from two major referral health centers for newborn screening between 2014 and 2016. Samples from thirty patients with a definite diagnosis of severe combined immunodeficiency, X-linked agammaglobulinaemia (XLA), ataxia telangiectasia (AT) and twenty-five healthy controls were used for determining the cutoffs. Samples from other patients including those with IgA deficiency (IgAD), Wiskott-Aldrich syndrome, Hyper-IgM syndromes (HIgM) and Dedicator of cytokinesis 2 (DOCK2) deficiency were considered as controls. TREC, KREC and beta-actin (ACTB) copy numbers were analyzed in all samples using triplex-quantitative real-time PCR. The diagnostic efficiency of this method was evaluated using ROC curve analysis. Results: Subsequent to statistical analysis, the cutoff values were calculated and obtained with a high sensitivity and specificity. Among the thirty cases with abnormal results (1.4%), twelve cases (0.6%) were retested and were subsequently shown to be in the normal range. Moreover, all of the positive, disease control patients were correctly identified. Conclusion: Determining cutoff levels with a high sensitivity and specificity for both TREC and KREC is essential for correctly identifying children with severe forms of PID in newborn screening programs, whilst minimising false positive results. Early diagnosis of patients with PID enables early implementation of appropriate preventative measures and definitive therapies such as hematopoietic stem cell transplantation. [ABSTRACT FROM AUTHOR]

One of the components of NADPH oxidase is p47‐phox, encoded by NCF1 gene. This study aims to find new genetic changes and clinical features in 38 Iranian patients with autosomal recessive chronic granulomatous disease (AR‐CGD) caused by NCF1 gene defect. Patients who had abnormal NBT and DHR‐1,2,3 assay with loss of p47‐phox in Western blotting were included in this study. After recording demographic and clinical data, PCR amplification was performed followed by direct sequencing for all exons and exon‐intron boundaries. The most common form of CGD in Iran was AR‐CGD due to consanguinity marriages. Among patients with AR‐CGD, NCF1 deficiency was found to be more common than other forms. Cutaneous involvements (53%), pulmonary infections (50%) and lymphadenopathy (29%) were more prevalent than other clinical manifestations of CGD. Mutation analysis of NCF1 gene identified five different mutations. Homozygous delta GT deletion (c.75_76delGT) was the most frequent mutation and was detected in more than 63% of families. Six families had a nonsense mutation in exon 7 (c.579G > A). Two novel mutations were found in exon 4 in two families, including a missense mutation (c.328C > T) and a nine‐nucleotide deletion (c.331_339delTGTCCCCAC). Genetic detection of these mutations may result in early diagnosis and prevention of possible complications of the disease. This could be useful for timely decision‐making for haematopoietic stem cell transplantation and for carrier detection as well as prenatal diagnosis of next children in the affected families. Our findings might help to predict outcomes, raise awareness and help effective treatment in these patients. [ABSTRACT FROM AUTHOR]

Objective: Nowadays genetic information is need to diagnose, treat, and generally manage different diseases. So genetic testing of immunodeficiency (PID) disorders is also of importance, especially for decisive diagnosis as well as bone marrow transplantation. Registration of information based on genetics has potential benefits for families possessing individuals with that disease for both present and future. This registry aims to collect genetics information about PID patients and their families' members. Materials and Methods: All of the Iranian patients suspected to PID who referred to immunology, asthma and allergy research institute (IAARI) between 2007 and 2016have been investigated regarding to PID. After taking informed consent, screening tests for PID were performed completely. Genetics data of individuals after definite diagnosis were enrolled in the genetic registry. Results: Genetic testing of 15 different PID diseases has been performed for more than 450 cases and collected in this registry. Of these 258 cases and their families had decisive mutation and genetic diagnosis. The rate of diseases was: Chronic Granulomatous Disease 22.17%, Severe Combined Immunodeficiency 21.33%, Wiskott-Aldrich Syndrome 10.46%, Congenital Neutropenia 8.36%, Leukocyte Adhesion Deficiency 7.53%, Griscelli 5.85%, Hemophagocytic Lymphohistiocytosis 5.23%, Angioedema 5.23%, Cystic Fibrosis 3.76%, Hyper IgE 2.92%, Hyper IgM 2.92%, X-linked Agammaglobulinemia 2.72%, Hermansky-Pudlak Syndrome 0.83%, Ataxia-Telangiectasia 0.41% and WHIM Syndrome 0.20%. Conclusions: This registry can be valuable for making aware of patients including decisive diagnosis, genetic counseling, carrier detection and prenatal diagnosis. In addition, this is a bank of data for future research. [ABSTRACT FROM AUTHOR]

Background: Chronic granulomatous disease (CGD) is a rare primary immunodeficiency (approximately 1 case in 200.000-250.000 newborns) caused by defect in one of the components of nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase enzyme (membrane-bound glycoproteins gp91, and cytosolic subunits p47-phox, p67-phox, p40-phox and p22-phox). This leads to defective intracellular killing by phagocyte cells. One of the components of NADPH-oxidase is p22-phox which is encoded by CYBA gene. In this study were identified new genetic changes of CYBA in 22 Iranian patients with autosomal recessive-CGD (AR-CGD). Methods: Twenty two CGD patients based on defect in NADPH oxidase activity (by NBT slide test and DHR-123 assay) and their demographic data and clinical histories were entered this study. They were referred to Immunology, Asthma and Allergy Research Institute (IAARI) for diagnosis and treatment. All patients had p22-phox deficiency based on western blotting. Genomic DNA was extracted from peripheral blood mononuclear cells (PBMCs) and PCR followed by direct sequencing to find p22-phox mutations. Results: Mutation analysis of CYBA revealed 12 different mutations, including three novel mutations: one deletion in exon 1, and two transitions in exon 3 [c.136G>A (p.Gly46Ser)], and exon 6 [c.388C>T (p.Gln130X)]. Conclusions: Three new mutations of CYBA gene in 4 of 22 Iranian patients with AR-CGD were found. These 3 novel mutations can partly complete the data base of Human Gene Mutation Database (HGMD). It can also be helpful for further prenatal diagnosis of next children in the affected families. Also it will be useful for timely decision making in bone marrow transplantation as treatment for CGD patients. [ABSTRACT FROM AUTHOR]