IgE-mediated allergic responses associated to Ailanthus altissima pollen using an animal model.

BACKGROUND: Murine models have been widely used in the study of allergy as sensitized mice can produce IgE and/or IgG1in response after the injection of an antigen/adjuvant combination. Ailanthus altissima pollen (AAP) has been recently reported as an emerging aeroallergen in Iran. So far, several AAP candidate allergens by the screening of allergen-specific IgE in the sera from AAP sensitized patients in Iran. OBJECTIVE: The aim of the present study was to detect and compare the allergens eliciting an IgE response in a mouse model, and in human, using pollen extract of A. altissima and an immunoproteomics based approach. METHODS: The pollen proteins were extracted in phosphate-buffered saline (PBS). Thirty male BALB/c mice were randomly divided into two groups of AP extract sensitized and sham that respectively received AAP PBS extract and a PBS control by intraperitoneal injections at regular intervals. The optimized AAP protein extracts were analyzed using 2D-gel electrophoresis and were subsequently confronted to pooled sera of sensitized mice. RESULTS: Two-D gel electrophoresis of AAP extract allowed the separation of 125 protein spots distributed in a wide range of pI and molecular masses. Two-DE immunoblotting using pooled sera of sensitized mice led to the detection of 14 IgE reactive spots with molecular masses ranging from 12 to 40-42kDa. CONCLUSION: The results do not correlate with our previous analyses using human AAP-sensitized sera. These findings reflect some differences in the sIgE reactivity to allergenic proteins in animal models.

Are the most common food allergens in an Iranian atopic population compatible with worldwide reports? A systemic review and meta-analysis with molecular classification of frequent allergens.

BACKGROUND: Undesirable immunological responses to alimentary allergens are one of the hallmarks of atopic diseases. The prevalence of common food allergens is dissimilar among different communities with distinct nutritional habits and genetic characteristics. AIM: To assess the prevalence of the most common food allergens in Iran, using different reliable studies. METHODS: All studies determining sensitization to common food allergens that were indexed in PubMed, Web of Science, Google Scholar, ProQuest, Scopus, Iran Medex, and Magiran were included in this review. To perform a meta-analysis, STATA 14 and metaprop command was applied. A logistic-normal random-effects model with Freeman-Tukey double arcsin transformation was applied to combine the findings of different studies and evaluate their heterogeneity. Random pooled estimate (ES) (pooled prevalence), 95% confidence interval (95% CI) and p-value were determined. RESULTS: A total of 23 studies with data from a total of 6126 children and adults met the inclusion criteria for entering this meta-analysis. The respective pooled prevalence of a positive family history of allergy and positive specific IgE to at least one food allergen was 72% (95% CI: 66-77%) and 41% (95% CI: 33-49%), respectively. Our results in the total population revealed that allergic sensitization to egg yolk, cow's milk (CM), egg white, and wheat were 25% (95% CI: 16%-35%), 24% (95% CI: 19-29%), 23% (95% CI: 18%-28%), and 9% (95% CI: 6%-14%), respectively. Walnut, peanut, and soybean sensitization was detected in 23% (95% CI: 17%-31%), 23% (95% CI: 13%-33%), and 20% (95% CI: 12%-28%) of patients, respectively. Random pooled ES for sensitization to shrimp and fish was 32% (95% CI: 21-45%) and 12% (95% CI: 6-20%), respectively. The result of analysis in different age groups revealed that allergic sensitization to milk, egg white, and egg yolk declines in higher age groups; while shrimp sensitization increases in older patients. In patients with atopic dermatitis, egg white was the most frequent food allergen 29% (95% CI = 18-42%); while wheat was the least frequent 8% (95% CI = 4-14%). CONCLUSIONS: Considering the prevalence of different food allergens, the results of the current meta-analysis revealed that egg yolk and cow's milk had the second and third rate after shrimp, respectively. The high prevalence of sensitization to shrimp may be attributed to its high consumption in coastal areas and/or cross-reactivity of shrimp with some aeroallergens such as mites.

Three novel mutations in CYBA among 22 Iranians with Chronic granulomatous disease.

Chronic granulomatous disease (CGD) is a rare primary immunodeficiency caused by defect in one of the components of nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase enzyme. The enzyme is at least composed of membrane-bound subunits gp91-phox and p22-phox (also named cytochrome b558 ), and cytosolic ones p40-phox, p47-phox and p67-phox. A defect in the enzyme activity leads to impaired intracellular killing of phagocytic cells. The CYBA gene encoding p22-phox is located on chromosome 16q24. In this study, new genetic changes of CYBA gene in 22 Iranian patients with autosomal recessive-CGD (AR-CGD) were identified. Twenty-two patients with CGD were referred to Immunology, Asthma and Allergy Research Institute (IAARI) and enrolled in this study based on defect in NADPH oxidase activity, demographic data and clinical histories. 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 was performed to find p22-phox mutations. Mutation analysis of CYBA revealed 12 different mutations, including three novel mutations: one was deletion of exon 1, and two were point mutations in exon 3 (c.136G>A (p.Gly46Ser)), and exon 6 (c.388C>T (p.Gln130X)). Three new mutations of CYBA gene in four of 22 Iranian patients with AR-CGD were found. These three novel mutations can partly complete the database of Human Gene Mutation Database (HGMD) and other related ones. It can also be helpful for further prenatal diagnosis in the affected families. Given that currently bone marrow transplantation is considered to be the curative treatment for patients with CGD, finding mutations will also be useful for timely decision-making in bone marrow transplantation.

Clinical, Laboratory, and Molecular Findings for 63 Patients With Severe Combined Immunodeficiency: A Decade´s Experience.

BACKGROUND: Severe combined immunodeficiency (SCID) is a life-threatening pediatric disease. We report on the clinical evaluation, immunological assessment, molecular analysis, and outcomes of SCID patients in a tertiary referral center in Iran. METHODS: From January 2006 to December 2015, we performed a prospective cohort study in which initial screening and advanced immunological tests were carried out on patients suspected of having SCID. Genetic analysis was also performed to confirm the diagnosis. RESULTS: A total of 63 patients were diagnosed with SCID (43 male [68.3%]). The median age at onset and diagnosis and diagnostic delay were 40 and 110 and 60 days respectively. A total of 49 patients (77.8%) had a history of BCG vaccination, and of these, one-third experienced BCG-associated complications. The most common clinical manifestations were pneumonia, recurrent oral candidiasis, chronic diarrhea, and failure to thrive. Of the thirteen patients who underwent hematopoietic stem cell transplantation, 8 survived and 5 died before they could receive the transplant. Most patients (34.9%) were classified as having T-B-NK+ SCID and had a mutation in the RAG2 or RAG1 gene. CONCLUSIONS: Autosomal recessive SCID is the most common type in Iranian patients. Providing high-quality training to physicians and patients' families to reduce the diagnostic delay should be prioritized. It is also important to raise awareness of live vaccination and to expand stem cell donor registries to speed up the transplantation process.

Quantitative polymerase chain reaction for detection of human herpesvirus-7 infection in umbilical cord blood donors.

OBJECTIVE: Umbilical cord blood (UCB) has been a reasonable alternative to granulocyte colony-stimulating factor-mobilized peripheral blood or bone marrow, as a source of hematopoietic stem cells with a lower risk of graft-versus-host disease. In immunocompromised hosts after transplantation, the risk of viral infection in adults, especially with beta-herpesviruses such as human herpesvirus-7 (HHV-7), may be increased. This virus in immunocompromised patients can be reactivated from latency and converted to an active phase. Therefore, light-upon-extension real-time polymerase chain reaction (PCR) was developed to assess the prevalence and load of HHV-7 in the plasma and buffy coat of donors. METHODS: About 825 UCB samples under standard protocol from donors were collected. Then, DNA from plasma and buffy coat was extracted and quantitative real-time PCR was performed with light-upon-extension primers. RESULTS: Overall, HHV-7 was detected in 3.64% (30/825) of UCB donors. HHV-7 DNA was detected in 26 (3.2%) buffy coat samples (latent infection), and only 4 (0.48%) of them were positive for HHV-7 DNA in plasma samples (active infection); the mean HHV-7 viral load was 1.31 × 10(1) copies/mL in latent infection, and 1.94 × 10(5) copies/mL in active infection. CONCLUSIONS: We suggest that real-time PCR in plasma and buffy coat could be a useful method to detect active and latent HHV-7 infection in UCB donors and determine its role in subsequent transmission events.

Cytokine production by activated T cells in common variable immunodeficiency.

BACKGROUND: Common variable immunodeficiency (CVID) is the most common symptomatic antibody deficiency. It is characterized by hypogammaglobulinemia, increased susceptibility to recurrent infections, autoimmunity, and malignancies. OBJECTIVES: To determine whether patients with CVID have cytokine production defects after T-cell activation and to assess whether or not these are correlated with markers of severe disease. METHODS: Twenty-seven patients with CVID and 17 healthy volunteers were investigated. Peripheral blood mononuclear cells were cultured under standard conditions in the presence and absence of phytohemagglutinin. Subsequent cell proliferation and cytokine release were measured and compared between stimulated and unstimulated cells. RESULTS: A general enhancement in cytokine production was observed in both CVID patients and controls after stimulation. However, we detected a lower production of interferon-gamma in CVID patients than in controls (P = .026). A production defect for at least 1 cytokine was observed in 12 patients. Ten of these failed to generate protective titers in response to the polysaccharide vaccine, and the frequency of bronchiectasis in this group of patients was 91.7%. Cytokine release correlated strongly with cell proliferation. CONCLUSIONS: This study indicates that some CVID patients have T-cell proliferation and secretory defects and that these may be associated with severe manifestations of disease. Screening for such defects could permit more effective monitoring and therapeutic strategies for CVID patients.

A novel RAB27A mutation in a patient with Griscelli syndrome type 2.

Griscelli syndrome type 2 is a rare autosomal recessive primary immunodeficiency disease caused by a mutation in the RAB27A gene and characterized by oculocutaneous hypopigmentation and variable cellular immunodeficiency. We report the case of a 6-month-old infant with silvery hair, eyelashes, and eyebrows who was referred to our center because of fever and hepatosplenomegaly. Bone marrow studies indicated hemophagocytosis, whilst microscopic examination of the hair showed irregular agglomerations of pigment in hair shafts. Molecular analysis revealed a novel homozygous mutation in exon 5, namely, a single-base substitution (g.42996 A>G) leading to an amino acid change (S115G) and thus confirming the diagnosis of Griscelli syndrome type 2. Griscelli syndrome could be more common than thought, especially in regions with high rates of consanguinity. As the prognosis of disease is usually poor, prompt diagnosis and appropriate treatment are vital to avoid complications.

Instability of the structure and allergenic protein content in Arizona cypress pollen.

BACKGROUND: The allergenic characteristics of pollen and their levels of expression may vary depending on the plant species, the degree of maturation and the influence of environmental factors such as climate and atmospheric pollution. The objective of this survey was the comparison of the structure and allergenic protein content in Arizona cypress (Cupressus arizonica, CA) pollen collected just after microsporangia dehiscence and 2 weeks later in urban areas. METHODS: The morphology and structure of pollen were examined by scanning electron microscopy. Pollen protein content was quantitatively and qualitatively investigated by Bradford protein assay, SDS-PAGE and densitometric analysis respectively. Fifteen allergic subjects, according to their clinical history of seasonal rhino-conjunctivitis and bronchial asthma have been selected for skin prick testing and ImmunoCap using CA standard allergen and for immunoblotting using extracts of CA mature pollen collected from Tehran. RESULTS: After 2 weeks, numerous cracks and collapses appeared in pollen surfaces. Western blotting performed by using extracts of pollen collected from Tehran, revealed that sera-specific immunoglobulin E of all allergic subjects reacted to a 35 kDa protein. The presence of this new major allergen and the decrease of Cup a 1 provide reliable explications about the low efficiency of standard commercial allergens in the diagnosis of the CA pollen allergy in Tehran. CONCLUSION: The instability of the pollen structure and protein content affects CA pollen allergenic properties. This study also suggests that to optimize CA standard allergen preparations, the eventual variability of pollen allergenic components have to be considered for each region.

Adverse drug reactions in an Iranian department of adult infectious diseases.

This study aimed to assess the frequency and severity of adverse drug reactions (ADRs) (WHO definition) in hospitalized adult patients in an infectious diseases referral ward in Tehran. Of 281 patients evaluated over 6 months, a total of 170 suspected ADRs were reported among 101 patients (35.9%). The most commonly affected organ system was gastrointestinal (47.5%), and the most common class of drugs responsible was anti-infectives (93.1%). ADRs were high among HIV-positive patients (82.9%), mainly due to anti-tuberculosis drugs. Attention to appropriate prescription of drugs is required with more careful clinical and laboratory monitoring of patients.

Molecular diagnosis of X-linked chronic granulomatous disease in Iran.

Chronic granulomatous disease (CGD) is an inherited disorder of pathogen killing by phagocytic leukocytes caused by mutations in NADPH oxidase subunits. Patients with CGD have life-threatening bacterial and fungal infections. Children's Medical Center at Tehran University is the referral center for immunodeficiency in Iran. During 2 years of study, 11 non-consanguineous families with clinically diagnosed CGD were referred to this center. In functional assays performed on neutrophils from affected children and their mothers; no activity or strongly decreased oxidase activity was detected in the patients' cells. In oxidase tests that scored this activity on a per-cell basis, a mosaic pattern was detected in the neutrophils from all 11 mothers. Western blot analysis revealed an X91 degrees phenotype in all patients. Mutation screening in the CYBB gene encoding gp91(phox) by SSCP analysis followed by sequencing showed nine different mutations, including two novel mutations. The present survey is the first study aimed to analyze the clinical features and the molecular diagnosis of X-CGD in Iranian patients.

Alterations in humoral immunity in relatives of patients with common variable immunodeficiency.

BACKGROUND AND OBJECTIVES: It has been reported that there is a high prevalence of immunodeficiency and autoimmunity in relatives of patients with common variable immunodeficiency (CVID). The aim of this study was to determine the prevalence of immunoglobulin deficiency in relatives of patients with CVID in Iran, where there is a high rate of consanguineous marriage. METHODS: A descriptive study was undertaken in 64 family members of 23 unrelated CVID patients. The group contained 17 fathers, 18 mothers, 18 sisters, 9 brothers, and 2 children. Serum immunoglobulin levels were measured by nephelometry. Immunoglobulin (Ig) G subclass levels were measured in a subgroup of 36 individuals. Serum IgA levels were confirmed by enzyme-linked immunosorbent assay for subjects with suspected IgA deficiency. RESULTS: The rate of consanguineous marriage in families containing relatives with antibody deficiencies was significantly higher than in those families in whom relatives did not have immune deficiencies. IgA deficiency was observed in 2 relatives of patients with CVID. Also CVID was observed in 2 family members. In 3 fathers and 1 brother, IgM levels were lower than normal. Three relatives had IgG4 deficiency and 1 person had combined IgG4 and IgG2 deficiency. Twenty percent of the relatives had hypogammaglobulinemia (including IgA deficiency, CVID, decreased levels of IgM, and IgG subclass deficiencies). CONCLUSION: In our study, alteration in humoral immunity in relatives of CVID patients was higher than previously reported, and this could be attributed to the high rate of consanguineous marriage in Iran. Since the family members of CVID patients are at high risk of hypogammaglobulinemia, it is advisable that they be evaluated for immunodeficiency disorders and monitored throughout their lifetimes.

T- helper 1 and 2 cytokine assay in patients with common variable immunodeficiency.

BACKGROUND: Common variable immunodeficiency (CVID) is a heterogeneous group of disorders, characterized by decreased immunoglobulin levels in serum and increased susceptibility to recurrent infections, autoimmunity, and malignancy. The pathogenesis of CVID is still unknown. OBJECTIVES: This study was performed to investigate T-helper (T(H)) 1 and 2 cytokine levels in patients with CVID. MATERIAL AND METHODS: Twenty-four cases of CVID were studied. Cytokine levels of interleukin (IL)-2, IL-4, IL-10, and interferon gamma were measured in the serum of the patients and compared with those of healthy controls. RESULTS: T(H)2 cytokine levels (IL-4 and IL-10) were significantly higher in the patient group than in the controls (median: 64.5 vs 0.0 pg/mL, P=.016 for IL-4 and 321.1 vs. 0.0 pg/mL, P=.001 for IL-10). However, there were no significant differences in T(H)1 cytokines between the 2 groups (median, 116.5 vs. 104.5 pg/mL, P=.22 for IL-2 and 50.5 vs. 42.3 pg/mL, P=.32 for IFN-gamma). CONCLUSIONS: Increased levels of IL-4 and IL-10 could indicate high activation of T(H)2 lymphocytes in this group of patients and consequently supports the concept of a bias towards T(H)2-type responses.

Quantification of Active and Latent Form of Human Cytomegalovirus Infection in Umbilical Cord Blood Donors by Real-Time PCR.

BACKGROUND: Umbilical cord blood (UCB) is believed to be a highly valuable source of hematopoietic stem cells for transplantation. Objective: To investigate the prevalence of active and latent human cytomegalovirus (CMV) infection in UCB donors in Iranian population. METHODS: A total of 825 UCB samples was collected under standard procedures and analyzed for the presence of CMV DNAs in buffy coat (latent infection) and plasma (active infection). DNA was extracted from buffy coat and plasma samples separately and tested with quantitative real-time PCR. All positive samples were checked by ELISA for IgG and IgM anti-CMV antibody. RESULTS: Latent CMV infection was detected in 17 (2%) buffy coat samples with a low level of viral load, which indicated the presence of latent viral infection in donors. None of the plasma samples were found positive for CMV DNA reflecting no active infection. In the 17 positive samples, CMV viral load was 91-104 (mean: 100) copies/mL. All samples positive for viral DNA were also found positive for CMV IgG antibody by ELISA. No CMV IgM antibody was detected in positive samples. CONCLUSION: CMV is still the most important virus that infects hematopoietic stem cells and could be dangerous, especially for immunocompromized transplant recipients. We therefore suggest using real-time PCR for the detection and quantification of the viral DNA in buffy coat and plasma of UCB donors. PCR of plasma for detection of CMV and antibody assay for CMV infection add no more sensitivity for the detection of latent CMV infection in UCB donors.

Pulmonary complications in primary hypogammaglobulinemia: a survey by high resolution CT scan.

BACKGROUND: Primary hypogammaglobulinemia disorders are a group of heterogeneous immunodeficiency syndromes with an increased susceptibility to pulmonary complications. METHODS: The aim of this study was to evaluate the extent of lung abnormalities in primary hypogammaglobulinaemic patients by high resolution computed tomography (HRCT) scan and pulmonary function test (PFT). HRCT and PFT were performed in 22 Iranian patients with primary hypogammaglobulinemia. RESULTS: Pathological bronchial findings were observed in thirteen patients: three patients showed only peribronchial thickening and the remaining ten patients suffered from both bronchiectasis and peribronchial wall thickening. Mild type of bronchiectasis and peribronchial wall thickening were the most common type, predominantly observed in the right middle and both right and left lower lobe segments of lungs. Although bullae were not found, emphysema, air-trapping, and collapse/consolidation were observed in two patients. Bronchial involvement was mostly limited to 1 up to 5 bronchopulmonary segments; only one HRCT indicated bronchial involvement in more than nine bronchopulmonary segments. Pathological bronchial findings mostly observed in the proximal bronchi; meanwhile the involvement of the distal bronchi was less common. Decreasing FEVI and FVC were observed in 65% and 55% of patients, respectively. There was a significant correlation between the HRCT score and the predicted values by PFT. The delay of diagnosis in patients with bronchiectasis was significantly higher than those without bronchiectasis. CONCLUSIONS: It seems that the majority of hypogammaglobulinaemic patients suffer from the mild type of bronchiectasis, which is mostly observed in the proximal bronchi of the lower lobe segments. The delay of diagnosis plays an important role in the occurrence of this complication in these patients.

Long-term outcomes of fludarabine, melphalan and antithymocyte globulin as reduced-intensity conditioning regimen for allogeneic hematopoietic stem cell transplantation in children with primary immunodeficiency disorders: a prospective single center study.

Reduced-intensity conditioning (RIC) has offered many primary immunodeficiency disorder (PID) patients who are ineligible for myeloablative regimens a chance of cure. However, the beneficial role of RIC was questioned following reports suggesting higher chance of rejection and lower symptom resolution rate in mixed chimerism settings. Forty-five children affected by PIDs with a median age of 21 months underwent allogeneic hematopoietic stem cell transplantation in our institute from 2007 to 2013. All patients received an identical RIC regimen. Forty-one patients had successful primary engraftment (91%). Of the successful engraftments, 80% (n=33) had stable full donor chimerism at last contact. Overall, eleven transplant-related mortalities were reported including five patients due to sepsis, three children due to grade IV acute GvHD, two due to chronic GvHD and one patient due to sepsis after primary graft failure. The median post-transplantation follow-up of deceased patients was 55 days. Five-year overall survival and disease-free survival was 75.6% and 68.89%, respectively. All surviving patients with successful engraftment became disease free, regardless of having full or mixed chimerism. Our study suggests that RIC regimen provides satisfactory rates of successful engraftment and full chimerism. Furthermore, patients with mixed chimerism were stable in long-term follow-up and this chimerism status offered the potential to resolve symptoms of immunodeficiency.

Serum concentration of selenium in healthy individuals living in Tehran.

OBJECTIVE: To investigate whether daily diet provides adequate selenium intake in healthy men and women living in Tehran, Iran. METHOD: Serum level of selenium was determined in 184 healthy individuals of both genders. The samples were divided into two age groups, adults and children, for analysis. The serum level of selenium was determined using hydride generation and flame atomic absorption spectroscopy. RESULTS: The mean and standard deviation of serum selenium levels in children (1-16 years) was 84.3 +/- 11 microg/l and there was no significant difference between genders in this group. In adults (older than 16 years) the mean serum selenium level was 100.6 +/- 13 SD microg/l; among women the mean was 93.9 +/- 14 SD microg/l and among men it was 102.2 +/- 12 SD microg/l. The mean selenium level in men was higher than in women and data analysis showed a significant difference between them (p < 0.005). There was also a positive correlation between higher selenium serum concentration and age in men (P < 0.001). Daily intake of selenium in men and women was calculated to be 67 microg and 62.1 microg respectively. CONCLUSION: Our results show that the serum concentration of selenium in an Iranian population is similar to other nationalities in the Middle East, particularly Saudi Arabia.

Distribution of primary immunodeficiency disorders diagnosed in the Children's Medical Center in Iran.

Primary immunodeficiency disorders include a variety of diseases that render patients more susceptible to infections. To determine the percentage of different primary immunodeficiency disorders diagnosed in the Children's Medical Center Hospital affiliated to Tehran University of Medical Sciences in Iran, we retrospectively reviewed the charts of the patients being referred to our hospital for immunologic evaluation of recurrent infections during a 20 year period. Among these patients, antibody deficiencies were the most frequent ones and were found in 52.6% of patients (n = 130). T-cell disorders, phagocytic disorders and complement deficiencies were found to be present in 24.69% (n = 61). 22.2% (n = 55) and 0.4% (n = 1) respectively. On the whole, common variable immunodeficiency was the most frequent disorder (n = 65), followed by ataxia telangiectasia (n = 39), X-linked agammaglobulinemia (n = 33), chronic granulomatous disease (n = 29) and selective IgA deficiency (n = 20). This study reveals that antibody deficiencies are the most common type of disorders as shown in other studies. A comparative study shows some differences between our results and other registries. This article also indicates that immunodeficiency disorders should be considered in patients with recurrent infections.