Comparison of Common Monogenic Defects in a Large Predominantly Antibody Deficiency Cohort.

BACKGROUND: Predominantly antibody deficiencies (PADs) are the most common primary immunodeficiencies, characterized by hypogammaglobulinemia and inability to generate effective antibody responses. OBJECTIVE: We intended to report most common monogenic PADs and to investigate how patients with PAD who were primarily diagnosed as suffering from agammaglobulinemia, hyper-IgM (HIgM) syndrome, and common variable immunodeficiency (CVID) have different clinical and immunological findings. METHODS: Stepwise next-generation sequencing and Sanger sequencing were performed for confirmation of the mutations in the patients clinically diagnosed as suffering from agammaglobulinemia, HIgM syndrome, and CVID. RESULTS: Among 550 registered patients, the predominant genetic defects associated with agammaglobulinemia (48 Bruton's tyrosine kinase [BTK] and 6 µ heavy chain deficiencies), HIgM syndrome (21 CD40 ligand and 7 activation-induced cytidine deaminase deficiencies), and CVID (17 lipopolysaccharides-responsive beige-like anchor deficiency and 12 atypical Immunodeficiency, Centromeric instability, and Facial dysmorphism syndromes) were identified. Clinical disease severity was significantly higher in patients with µ heavy chain and CD40 ligand mutations compared with patients with BTK (P = .003) and activation-induced cytidine deaminase (P = .009) mutations. Paralysis following live polio vaccination was considerably higher in patients with µ heavy chain deficiency compared with BTK deficiency (P < .001). We found a genotype-phenotype correlation among patients with BTK mutations regarding clinical manifestation of meningitis and chronic diarrhea. Surprisingly, we noticed that first presentations in most patients with Immunodeficiency, Centromeric instability, and Facial dysmorphism were respiratory complications (P = .008), whereas first presentations in patients with lipopolysaccharides-responsive beige-like anchor deficiency were nonrespiratory complications (P = .008). CONCLUSIONS: This study highlights similarities and differences in the clinical and genetic spectrum of the most common PAD-associated gene defects. This comprehensive comparison will facilitate clinical decision making, and improve prognosis and targeted treatment.

DNAH11 Localization in the Proximal Region of Respiratory Cilia Defines Distinct Outer Dynein Arm Complexes.

Primary ciliary dyskinesia (PCD) is a recessively inherited disease that leads to chronic respiratory disorders owing to impaired mucociliary clearance. Conventional transmission electron microscopy (TEM) is a diagnostic standard to identify ultrastructural defects in respiratory cilia but is not useful in approximately 30% of PCD cases, which have normal ciliary ultrastructure. DNAH11 mutations are a common cause of PCD with normal ciliary ultrastructure and hyperkinetic ciliary beating, but its pathophysiology remains poorly understood. We therefore characterized DNAH11 in human respiratory cilia by immunofluorescence microscopy (IFM) in the context of PCD. We used whole-exome and targeted next-generation sequence analysis as well as Sanger sequencing to identify and confirm eight novel loss-of-function DNAH11 mutations. We designed and validated a monoclonal antibody specific to DNAH11 and performed high-resolution IFM of both control and PCD-affected human respiratory cells, as well as samples from green fluorescent protein (GFP)-left-right dynein mice, to determine the ciliary localization of DNAH11. IFM analysis demonstrated native DNAH11 localization in only the proximal region of wild-type human respiratory cilia and loss of DNAH11 in individuals with PCD with certain loss-of-function DNAH11 mutations. GFP-left-right dynein mice confirmed proximal DNAH11 localization in tracheal cilia. DNAH11 retained proximal localization in respiratory cilia of individuals with PCD with distinct ultrastructural defects, such as the absence of outer dynein arms (ODAs). TEM tomography detected a partial reduction of ODAs in DNAH11-deficient cilia. DNAH11 mutations result in a subtle ODA defect in only the proximal region of respiratory cilia, which is detectable by IFM and TEM tomography.

Long-term evaluation of a historical cohort of Iranian common variable immunodeficiency patients.

OBJECTIVES: Common variable immune deficiency (CVID) is the most frequent form of symptomatic primary immunodeficiency disease, characterized by hypogammaglobulinemia, recurrent infections and increased predisposition to autoimmunity and malignancies. The aim of this study was to reconsider important points of previously performed studies on Iranian CVID patients diagnosed and followed from 1984 to 2013. METHODS: Diagnosis was made using approved criteria including reductions of serum levels of immunoglobulins and exclusion of well-known single gene defects in individuals with an age >4 years and evidence of specific antibody deficiency. RESULTS: Detailed information on demographic data, survival rates, clinical phenotypes, immunologic and genetic data and treatment of 173 patients are provided. The early onset presentation (74.5%) and rate of consanguineous marriage (61.2%) were considerably higher in our cohort. Our study revealed clinically related correlations regarding consanguinity, the population of naïve CD4(+) T cells and switched-memory B cells, cytokine levels and special genetic factors (including HLA and AID genes). CONCLUSION: Despite current efforts, more comprehensive studies are needed, especially for classification and investigation of the genetic background and prognostic factors for patients with CVID in order to better managment and followup of patinets.

Clinical and genetic features in patients with cystic fibrosis in southwestern iran.

OBJECTIVE: Cystic fibrosis (CF) is a common autosomal recessive genetic disease caused by a mutation in the CF transmembrane conductance regulatory (CFTR) gene. This study attempted to identify the most common CFTR mutations and any correlations between certain mutations and the clinical presentation of the disease in CF patients in southwestern Iran. METHODS: Twenty nine common CFTR gene mutations were examined in 45 CF patients. FINDINGS: Chronic cough, intestinal obstruction, dehydration, heat exhaustion and steatorrhea were the most common early clinical symptoms among our patients. The most common mutation was ΔF508, with an allele frequency of 21%. The homozygous ΔF508 mutation was observed in eight patients (18%), and three patients (7%) were ΔF508 carriers. The 2183AA > G mutation was observed in four patients, one of whom was also a ΔF508 carrier. The R1162X mutation was detected in two patients. The G542X, R334W and N1303K mutations were detected each in one patient, the first of whom was also a ΔF508 carrier. CONCLUSION: Out of 45 patients, 27 (60%) had none of the CFTR gene mutations we tested for. The most frequent mutations in southwestern Iranian patients with CF should be identified by sequencing the entire CFTR gene in order to optimize the design of a diagnostic kit for common regional mutations.

Interstitial lung disease in a patient with chronic granulomatous disease.

BACKGROUND: Chronic granulomatous disease (CGD) is an inherited phagocytes defect, characterized by defects of NADPH-oxidase and inability of bacterial killing, which leads to recurrent life-threatening infections. Respiratory problems, which are the major cause of morbidity in CGD, usually result from recurrent severe infections; however, vigorous inflammatory response could also cause respiratory diseases. CASE PRESENTATION: Herein, an 11 year-old patient with CGD is presented who suffered from chronic cough and dyspnea for 7 years. Considering the results of chest X-ray, high-resolution computed tomography, and pulmonary function test, the diagnosis of interstitial lung disease was made. CONCLUSION: Early recognition of manifestations associated with CGD and appropriate treatment could prevent further complications and reduce morbidity and mortality in this group of patients.

Survey of Mycoplasma pneumoniae in Iranian children with acute lower respiratory tract infections

OBJECTIVES: Mycoplasma pneumoniae is an atypical pathogen, which is one of the major causes of lower respiratory tract infections (LRTIs) worldwide. This study was performed to determine the role of M. pneumoniae in acute LRTIs in children, who were referred to main pediatric hospitals in Shiraz, Iran, with the diagnosis of LRTI. Polymerase chain reaction method on a throat-swab specimen was utilized to detect M. pneumoniae. RESULTS: One hundred patients with acute LRTIs were investigated in this study. There were 10 positive PCR for M. pneumoniae (10 percent), including 6 of 62 hospitalized patients and 4 of 38 outpatients. All patients with LRTIs due to M. pneumoniae had cough. Fever, flu like symptoms, dyspnea, pulmonary rales, wheezing, and conjunctivitis were other common signs and symptoms. CONCLUSIONS: The percentage of cases with M. pneumoniae infection in our population is similar to the reported in other parts of Asia. Precise and early detection of pathogen and appropriate antibiotic therapy are the key points in management of patients with LRTIs.

Pulmonary function test in transfusion-dependent ß-thalassemia major patients: a pilot study.

Lung involvement is one of known complications of thalassemia. The aim of this study was to determine predominant type of pulmonary dysfunction and its relationship to iron overload in ß-thalassemia children. Fifty thalassemia major children with treatment of regular blood transfusion and desferrioxamine participated in the study. Thirty-three boys and 17 girls (median age 12.5 years) with ß-thalassemia enrolled in the study. Other information including body mass index, hematocrit, and the number of years of blood transfusion were recorded. Serum ferritin level and hematocrit were 3346 ± 1667 mg/dL and 27.7 ± 2, respectively. Pulmonary function tests were performed in all subjects for detecting pulmonary dysfunction. Thirty-five patients (70%) with thalassemia had abnormal result of spirometry. Obstructive airway disease based on reduced forced expiratory volume in 1 second (FEV(1)) and FEV(1)/forced vital capacity (FVC) ratio <80% was detected in 4 patients (8%). Six patients (12%) showed restrictive pattern, as defined by a reduction FVC <80% and FEV(1)/FVC ratio ≥80%. In this study, small airway involvement based on presence of forced expiratory flow (FEF(25%-75%)) <60%, FEV(1)/FVC ratio >70%, and FVC >80% was detected in 25 subjects (50%). Decreased values of peak expiratory flow rate (PEF) were detected in 23 (46%) and low FEV(1) in 10 (20%) subjects. There was no significant correlation between abnormal pulmonary function test and serum ferritin level in children with thalassemia. This study showed small airway disease was predominant abnormality in thalassemia patients, although additional larger studies are needed to evaluate underlying mechanisms and validate these findings.

A case of hypogammaglobulinemia with enteroviral meningoencephalitis, associated with increased adenosine deaminase in cerebrospinal fluid.

We describe the development of enterovirus meningoencephalitis associated with increased adenosine deaminase in cerebrospinal fluid of a 12-year-old boy, a known case of hypogamaglobulinemia despite monthly replacement of IVIg.The patient was referred to our center with fever, headache and vomiting for 10 days. CSF analysis was compatible with aseptic meningoencephalitis but high CSF protein (>200mg/dl) and high level of adenosine deaminase in CSF (30IU/L) were against the diagnosis of simple viral meningoencephalitis. Nested PCR of CSF for entrovirus was positive. Treatment with daily high-dose IVIg was commenced, with significant clinical improvement. For patients with increased ADA and lymphocytic pleocytosis in CSF, differential diagnoses should include enteroviral meningitis. Antibodies, although crucial, cannot on their own prevent enteroviral infection in some hypogamaglbulinemic patients.

Clinical and laboratory findings in hyper-IgM syndrome with novel CD40L and AICDA mutations.

BACKGROUND: Hyper-immunoglobulin M (HIGM) syndromes are a heterogeneous group of primary immunodeficiency disorders, characterized by recurrent infections associated with decreased serum levels of immunoglobulin G (IgG) and IgA and normal to increased serum levels of IgM. These patients have immunoglobulin class switch recombination defects, caused by mutations in several genes. METHODS: In order to investigate clinical and immunological manifestations of HIGM in Iran, 23 Iranian patients with an age range of 5 months to 35 years, who were followed up over a period of 17 years, were studied. Fourteen of the 23 patients were screened for CD40L, AICDA, UNG, and CD40 gene mutations, using polymerase chain reaction followed by direct sequencing. RESULTS: All patients, except one, initially presented with infectious diseases; the most common manifestations were respiratory tract infections. Six different CD40L mutations were identified, five were novel, one splicing (IVS1+2T>C), three missense (T254M, G167R, L161P), and two frame shift deletions (T29fsX36 and D62fsX79). In addition, one novel AICDA mutation (E122X) was detected. No mutation was found in six out of 14 analyzed patients. CONCLUSION: CD40L mutations comprise the most common type of immunoglobulin class switch recombination defects. There are several patients with HIGM phenotype, in which the underlying genetic defects remain to be identified. Other defects such as those in components of the mismatch repair mechanism could be potential candidates for the latter.

Agreement rate of skin prick test with tissue eosinophil count in patients with nasal polyps.

The pathogenetic mechanism of nasal polyps remains unknown, although allergy has been cited as an important factor in the etiology of nasal polyposis. Currently there is no definite histological criterion for differentiation of allergic from inflammatory nasal polyp. However, in a few studies, tissue eosinophil count has been used for this. This study aimed to find out the agreement rate of skin prick test and tissue eosinophil count in patients with nasal polyposis. Twenty five patients (18 males, 7 females) with nasal polyp were enrolled in this study. For each patient tissue sample from polyp material was taken for histopathological investigation. Moreover, skin prick test was performed for each patient using eleven common aeroallergens. Skin prick test was positive in 48% of the patients. Tissue eosinophil count of more than 50% was found in 75% of skin prick positive and in 69.2% of skin prick negative patients. Also tissue eosinophil count of more than 50% was found in 69.2% of patients with typical allergic symptoms as well as 75% of patients without allergic symptoms. No agreement was found between skin prick tests and tissue eosinophil counts in patients with nasal polyp. Also no difference was found between the tissue eosinophil counts in allergic and non allergic patients. Considering these results, it can be concluded that having a high tissue eosinophil count in patients with nasal polyp does not indicate that the polyp is allergic.

Magnetic resonance imaging (MRI) findings of joints in young beta-thalassemia major patients: fluid surrounding the scaphoid bone: a novel finding, as the possible effect of secondary hemochromatosis.

BACKGROUND: Beta-thalassemia major is a hereditary disease with developing hemochromatosis as a consequence of repeated blood transfusions. Degenerative arthropathy may be the first manifestation of primary hemochromatosis. OBJECTIVE: The aim of this study is to determine the possible effects of secondary hemochromatosis in joints of beta-thalassemia major patients, and to evaluate the sensitivity of magnetic resonance imaging (MRI) compared with plain x-ray, in the early diagnosis of hemochromatosis of the joints. METHODS: Thirty beta-thalassemia major patients were randomly enrolled. A questionnaire regarding their history, careful physical examination, the result of their liver biopsies, and other necessary information were completed for these patients. X-ray and MRI of the left hand and wrist were taken in these patients and also in other 30 healthy people with the same age and sex as the case control group. RESULTS: The prevalence of minimal fluid surrounding the scaphoid bone in MRI was 23.3%, which is a novel and significant finding. Other interesting changes included hypo signal intensity (T1 and T2) in carpal, metacarpal, radius, and ulna bones. CONCLUSIONS: Regarding the availability of special health services for thalassemia patients, this imaging study can be used for the accurate detection of hemochromatosis in joints of thalassemia patients.

Frequency and clinical manifestations of patients with primary immunodeficiency disorders in Iran: update from the Iranian Primary Immunodeficiency Registry.

Primary immunodeficiency disorders (PID) are a heterogeneous group of diseases, characterized by an increased susceptibility to infections. A total of 930 patients (573 males and 357 females) are registered in Iranian PID Registry (IPIDR) during three decades. Predominantly antibody deficiencies were the most common (38.4%), followed by congenital defects of phagocyte number and/or function (28.3%), other well-defined immunodeficiency syndromes (17.7%), combined T- and B-cell immunodeficiencies (11.0%), complement deficiencies (2.4%), and diseases of immune dysregulation (2.3%). Common variable immunodeficiency was the most frequent disorder (20.8%), followed by chronic granulomatous disease, ataxia-telangiectasia, btk deficiency, selective IgA deficiency, and T-B-severe combined immunodeficiency. The frequency of other PID disorders was less than 50 in number (<5%). There is an increasing trend in recognition of more PID in the recent years. Construction of such registry is not only important for its epidemiological aspect but also for its role in increasing the physician's knowledge about such disorders.

A fast and easy nitroblue tetrazolium method for carrier screening and prenatal detection of chronic granulomatous disease.

BACKGROUND: Analysis of the functional activity of neutrophils is of great importance in the differential diagnosis of patients with recurrent bacterial infections. It has long been established that stimulated polymorphonuclear leukocytes reduce nitroblue tetrazolium. Application of a simple and reliable nitroblue tetrazolium method that clearly differentiates the chronic granulomatous disease patients and heterozygote carriers in some groups suspected to have chronic granulomatous disease was investigated. METHODS: This study consisted of 197 samples taken from 100 children (2 - 24-month-old) and 81 neonates (aged < 2 months) referred to our center either due to a suspected bacterial infection or suspected immunodeficiency. The sample also included 16 cord blood samples. Fifty healthy adult individuals were enrolled in this study and were diagnosed as normal control. Neutrophil reduction of nitroblue tetrazolium can be stimulated in vitro by protein kinase agonists such as phorbol myristate acetate, resulting in release of superoxide anion. RESULTS: Phorbol myristate acetate is an exceptionally powerful stimulant and when used in conjunction with glass adherence, caused nearly all normal neutrophils to become transformed and reduced nitroblue tetrazolium to formazan deposits. Of 197 blood samples, 9 were diagnosed as having unrelated chronic granulomatous disease and 7 were carriers of X-linked or autosomal recessive chronic granulomatous disease. The carriers had a range of 15 - 75% stimulated neutrophils. CONCLUSION: We have established a phorbol myristate acetate-stimulated nitroblue tetrazolium test for detection of chronic granulomatous disease patients, which clearly differentiates the chronic granulomatous disease patients from heterozygote carriers. The results in cord fetal blood indicate that this test may be used for antenatal diagnosis of affected boys, carrier females, and autosomal recessive variants of chronic granulomatous disease. The technique is simple, fast, inexpensive, and requires only a few microliters of blood.

The clinical and laboratory survey of Iranian patients with hyper-IgE syndrome.

In order to determine the clinical and laboratory findings of Iranian patients with presumed hyper-immunoglobulin E syndrome (HIES), the medical records of 22 patients from 21 unrelated families, who had been registered in the Iranian Primary Immunodeficiency Registry, were observed. The median age of patients at the time of first symptom and at the time of diagnosis was 1 month and 52.5 months, respectively, with a median diagnosis delay of 70 months. 13 families had consanguineous marriages. IgE level was higher than 2000 IU/ml in all patients, ranging from >2000 to 80,000 IU/ml. The most commonly occurring manifestations were: eczema and dermatitis, pneumonia, upper respiratory tract infections, cutaneous abscesses, diarrhoea, deep abscesses, and otitis media. Other less frequent manifestations were: mucocutaneous candidiasis, sinusitis, cutaneous ulcers, Molluscum contagiosum, herpetic keratitis, onychomycosis, conjunctivitis, septic arthritis, and meningitis. Five patients were complicated by bronchiectasis due to recurrent pneumonia and 5 patients died because of severe infections and malignancy. The HIES is a multisystem disorder that affects especially cutaneous, respiratory, skeletal and the immune system. Although HIES is a rare condition, the recurrent infections should always raise a suspicion, which deserves further evaluation for detecting the syndrome.

Consanguinity in primary immunodeficiency disorders; the report from Iranian Primary Immunodeficiency Registry.

PROBLEM: Primary Immunodeficiency Disorders (PiD) are a heterogeneous group of genetic disorders, with different modes of inheritance. This study was accomplished in order to determine the frequency of consanguineous marriages in the families of patients with PiD. METHOD: In this study, the records 515 Iranian PiD patients were reviewed during a 25-year period. RESULTS: The mean proportion of consanguineous marriages was 65.6% among PiD patients, while the overall rate was 38.6% in the country. The rate of consanguinity was 77.8% in cellular immunodeficiencies, 75.8% in combined immunodeficiencies, 72.5% in defects of phagocytic function, 58.6% in other immunodefiiencies, 54.1% in predominantly antibody deficiencies, and 50% in complement deficiencies. Moreover all patients with immunodeficiency associated with other diseases had consanguineous parents. Such marriages were most common in the parents of patients with Chediak-Higashi syndrome, severe combined immunodeficiencies, primary CD4 deficiency, ataxia-telangiectasia, selective IgG class deficiencies, chronic granulomatous disease, and Schwachman syndrome. CONCLUSIONS: It is important to inform the general population about the dangers of consanguinity, which is very common in some areas such as Iran. Premarital examination to avoid genetic diseases could be suggested, especially in a community where the rate of consanguineous marriage is high.

Hodgkin lymphoma developing in a 4.5-year-old girl with hyper-IgE syndrome.

The authors report a case of Hodgkin lymphoma developing in a 4.5-year-old female child with hyper-IgE syndrome. This is one of the few cases of malignancy reported in this syndrome. The patient had severe atopic dermatitis, asthma, recurrent pneumonia, recurrent skin infections, and growth retardation. Immunologic evaluation revealed a high level of immunoglobulin E (7000 IU/mL) and peripheral eosinophilia. She was found to have normal values for serum IgG, IgM, IgA, WBC chemotaxis, serum complement function and normal sweat chloride test. The development of fatal Hodgkin lymphoma in this patient with hyper-IgE syndrome may suggest an increased risk for developing premature malignancies in hyper-IgE syndrome, although the precise immunologic defect is still unknown.

Churg-strauss syndrome in an 8-year-old girl.

Churg-Strauss Syndrome (CSS) is a small vessel granulomatous vasculitis that characteristically affects the middle age group and is extremely rare in children. We describe an 8-year-old girl with a prolonged history of cough and eosinophilia that initially was labeled as pulmonary tuberculosis, but after complementary assays, associated with skin biopsy, eosinophilic fibrinoid vasculitis, CSS was eventually diagnosed. In this case, due to her young age and more prevalence of infectious lung diseases in our area, there was a few months delay for correct diagnosis and treatment. After meeting the CSS criteria the appropriate treatment was started and her condition improved.

Primary immunodeficiency in Iran: first report of the National Registry of PID in Children and Adults.

Epidemiological studies have shown wide geographical and racial variation in the prevalence and patterns of immunodeficiency disorders. To determine the frequency of primary immunodeficiencies (PID) in Iran, the Iranian Primary Immunodeficiency Registry (IPIDR) was organized in 1999. We extracted the patient's data, by using a uniform questionnaire from their hospital records. The diagnosis of patients was based on WHO criteria. By now, 440 patients with PID, who were observed during a period of 20 years, have been registered in our registry. Among these patients, the following frequencies were found: predominantly antibody deficiency in 45.9% of patients (n = 202), phagocytic disorders in 29.09% (n = 128), T-cell disorders in 24.31% (n = 107), and complement deficiencies in 0.68% (n = 3). Common variable immunodeficiency was the most frequent disorder (n = 98), followed by chronic granulomatous disease (n = 86), ataxia telangiectasia (n = 48), x-linked agammaglobulinemia (n = 45), selective IgA deficiency (n = 42), combined immunodeficiency (n = 15), and severe combined immunodeficiency (n = 14). This study revealed that antibody deficiencies is the most frequently diagnosed primary immunodeficiency disorder in our patients, which is similar to that observed in other registries. A comparative study shows some differences between our results and other registries.