Editorial

Innate lymphoid cells (ILCs) are identified as novel population of hematopoietic cells which protect the body by coordinating the innate immune response against a wide range of threats including infections, tissue damages and homeostatic disturbances. ILCs, particularly ILC2 cells, are found throughout the body including the brain. ILCs are morphologically similar to lymphocytes, express and release high levels of T-helper (Th)1, Th2 and Th17 cytokines but do not express classical cell-surface markers that are associated with other immune cell lineages.Three types of ILCs (ILC1, 2&3) have been reported depending upon the cytokines produced. ILC1 cells encompass natural killer (NK) cells and interferon (IFN)-g releasing cells; ILC2 cells release the Th2 cytokines, IL-5, IL-9 and IL-13 in response to IL-25 and IL-33; and ILC3 cells which release IL-17 and IL-22. ILC2 cells have been implicated in mucosal reactions occurring in animal models of allergic asthma and virus-induced lung disorders resulting in the regulation of airway remodeling and tissue homeostasis.There is evidence for increased ILC2 cell numbers in allergic responses in man but little is known about the role of ILCs in chronic obstructive pulmonary disease (COPD). Further understanding of the characteristics of ILCs such as their origin, location and phenotypes and function would help to clarify the role of these cells in the pathogenesis of various lung diseases.In this review we will focus on the role of ILC2 cells and consider their origin, function, location and possible role in the pathogenesis of the chronic inflammatory disorders such as asthma and COPD.

Aspergillus species (A flavus and A niger) are important sources of inhalant allergens. Current  diagnostic  modalities  employ  crude  Aspergillus  extracts  which  only  indicate  the source to which the patient has been sensitized, without identifying the number and type of allergens in crude extracts. We report a study on the identification of major and minor allergens of the two common airborne Aspergillus species and heterogeneity of patients’ IgE response to them.Skin prick tests were performed on 300 patients of bronchial asthma and/or allergic rhinitis and 20 healthy volunteers. Allergen specific IgE in patients’ sera was estimated by enzyme allergosorbent test (EAST). Immunoblots were performed to identify major/minor allergens of Aspergillus extracts and to study heterogeneity of patients’ IgE response to them.Positive cutaneous responses were observed in 17% and 14.7% of patients with A flavus and A niger extracts, respectively. Corresponding EAST positivity was 69.2% and 68.7%. In immunoblots, 5 allergenic proteins were identified in A niger extract, major allergens being 49, 55.4 and 81.5 kDa. Twelve proteins bound patients’ IgE in A flavus extract, three being major allergens (13.3, 34 and 37 kDa). The position and slopes of EAST binding and inhibition curves obtained with individual sera varied from patient to patient. The number and molecular weight of IgE-binding proteins in both the Aspergillus extracts varied among patients.These results gave evidence of heterogeneity of patients’ IgE response to major/minor Aspergillus allergens. This approach will be helpful to identify disease eliciting molecules in the individual patients (component resolved diagnosis) and may improve allergen-specific immunotherapy.

Inhalation of pollens from different species of Acacia is a common cause of respiratory allergy in tropical areas of the world. Acacia farnesiana is commonly used as street trees in towns and ornamental shade trees in parks and gardens throughout arid and semi-arid regions of Asia. This study aimed to produce and purify the A. farnesiana pollen profilin (Aca f 2) and evaluate its nucleotide sequence homology with profilins of common allergenic plants to predict allergenic cross-reactivity.Thirty-nine patients who were allergic to Acacia pollens were included in the study. Cloning of Acacia profilin-coding sequence was performed by polymerase chain reaction using primers from Acacia pollen RNA. The cDNA of Acacia pollen profilin was then expressed in Escherichia coli using pET-21b(+) vector and purified by metal affinity chromatography. Immunoreactivity of the recombinant Acacia profilin (rAca f 2) was evaluated by specific ELISA, immunoblotting, and inhibition assays.The coding sequence of the Acacia profilin cDNA was recognized as a 399-bp open reading frame encoding 133 amino acid residues. Eighteen patients (18/39, 46.15%) had significant specific IgE levels against Aca f 2. Immunodetection and inhibition assays indicated that purified Aca f 2 might be the same as that in the crude extract.Aca f2, the first allergen from A. farnesiana pollen, was identified as belonging to the family of profilins. The amino acid sequence homology analysis showed high cross-reactivity between Aca f 2 and other profilins from botanically unrelated common allergenic plants.

Interleukin (IL)-35 is a newly discovered suppressive cytokine and has been shown to alleviate  inflammatory  and  autoimmune  diseases.  The  purpose  of  this  study  was  to investigate immunomodulatory capacity of IL-35 in patients with allergic asthma.IL-35 mRNA expression levels in peripheral blood mononuclear cells (PBMCs) were detected  by  quantitative  real-time  PCR  (qPCR).  The  frequencies  of  cytotoxic  T  cells (Tc)1,Tc2  and  Tc17  cells  were  measured  by  flow  cytometry.  Plasma  levels  of  IL-35, interferon (IFN)-γ, IL-4, and IL-17 were examined by enzyme-linked immunosorbent assay (ELISA). The correlations between plasma IL-35 levels and Tc1, Tc2, and Tc17 cytokine production in allergic asthmatics (n = 25) and healthy controls (n = 12) were analyzed by Pearson’s test.IL-35 protein and mRNA expression levels were down-regulated in allergic asthmatics compared with healthy controls. The frequencies of Tc2 and Tc17 cells were significantly increased in patients with asthma, and the frequency of Tc1 cells did not differ between asthmatic patients and healthy controls. Similarly, plasma levels of IL-4 and IL-17 were significantly increased in asthmatic patients, while there was no difference in IFN-γ levels between allergic asthma patients  and  healthy  controls.  More importantly,  plasma  IL-35 protein levels were negatively correlated with the frequency of IL-4-producing CD8+ T (Tc2) cells and with the IL-4 level in patients with allergic asthma.Our results suggest that decreased circulating IL-35 levels could contribute to the pathogenesis of allergic asthma by regulating CD8+ T cells.

The SNP (rs11209026, Arg381Gln, R381Q) in the IL-23 receptor (IL23R) confers protection against multiple inflammatory diseases, representing one of the most significant human genetic polymorphisms in inflammatory diseases. We, therefore, investigated the association between IL-23 R R381Q gene polymorphism and asthma.This case-control study was performed on 209 patients, and 200 healthy controls. Using PCR-RFLP,  the  R381Q  variant  was  screened  in  the  IL-23R  gene  of  the  patients  and controls.Serum IgE levels were measured using ELISA technique. Eosinophil absolute count was done with Sesmex cell counter.Our results indicated that the genotype and allele frequencies of the IL-23R R381Q polymorphism is significantly different between asthmatic patients and control subjects (p<0.001; odd ratio= 0.266; 95%, CI=0.118-0.604. Moreover, the asthmatic patients had higher eosinophil count and total serum IgE levels than controls as expected (p<0.001).The present study suggested that R381Q polymorphism in IL-23 receptor may be a predisposing allele for asthma.

Respiratory virus infection is a major cause of asthma exacerbation. However, the underlying mechanisms of  this exacerbation  are unknown. Therefore,  to  determine  the mechanisms, we examined the effect of influenza infection in a murine model of asthma.Mice were divided into four groups: the phosphate-buffered saline (PBS), house dust mite (HDM), influenza, and HDM/influenza groups. The influenza group and the HDM/influenza group were infected with influenza A virus. We measured airway resistance (Penh value), examined the lung tissue for pathology, and analyzed the cells and cytokines in bronchoalveolar lavage fluid (BALF) by ELISA.At 50 mg/mL methacholine, the HDM/influenza group showed a significantly higher Penh value than the PBS, HDM, and influenza groups. The number of neutrophils in BALF was higher in the HDM/influenza group than in the HDM group. A significantly greater number of lymphocytes and macrophages were detected in the HDM/influenza group than in the HDM group. IFN-γ and IL-1β levels were higher in the HDM/influenza group than in the HDM group. IL-5 levels did not vary between the HDM and HDM/influenza groups, IL-10 was significantly lower in the HDM/influenza than in the HDM group. Chemokine (C-X-C motif) ligand 1 (CXCL1) and regulated upon activation, normal T cell expressed and secreted (RANTES) were higher in the HDM/influenza group than in the HDM group.In a murine model of asthma, influenza-induced airway inflammation appeared to becaused by simultaneous activation of neutrophilic and eosinophilic inflammation.

The purpose of this study was to assess the effect of preventive immunization on the incidence of allergies in Poland.18617 (53.8% female, 24.2% 6-7 years old, 25.4% 13-14 years old, 50.4% 20–44 years old) were selected by stratified cluster sampling method in 8 cities and 1 rural area. 4783 of whom underwent objective outpatient screening assessments. Study subjects were evaluated for any association  between  preventive  immunization  against  rubella,  measles,  typhoid  fever, smallpox and incidence of atopic dermatitis, allergic rhinitis, and asthma.There was no increased risk of allergy incidence in the majority of vaccinated subjects against rubella, measles, typhoid  fever, or smallpox (OR  from 0.42 (ppppp=0.005), varicella (OR=1.18; p=0.003); rhinitis and AR following vaccination against measles (respectively OR=1.22; pp=0.0002).  No  higher risk of allergic  diseases  was  demonstrated  in vaccinated individuals diagnosed by doctor in an outpatient setting.These data do not demonstrate a causal relationship between vaccinations and allergic conditions.

An increasing number of asthmatic patients are attracted by complementary and and alternative medicine (CAM). The aim of this study was to estimate the prevalence and describe the characteristics of CAM use by children with asthma in a paediatric allergy clinic in Istanbul, Turkey.The parents of asthmatic children were invited to participate in a cross-sectional survey study. Current asthma treatment, severity of asthma, emergency admittances and hospitalisations, education of parents, settlements, income of the family and parental use of CAM were investigated as predictors of CAM usage.Out of the 500 patients, 330 (66 %) had used CAM therapy; most popular modalities were herbal medicine (45 %), honey (41.6 %), grape syrup (37.2 %) and quail eggs (36.2 %). The most common used herbal medicine in the study group were linden (21.6%) and ginger (21.2 %). There was no significant difference in regard to the use of regular asthma treatment, the severity of asthma, the frequency of emergency admittance, hospitalisations due to asthma, education of parents and settlements between CAM users and non-CAM group. A significant inverse association was found in terms of family income and CAM usage. Parents’ own use of CAM was also associated with significant increase in the use of CAM.In conclusion; the prevalance of reported CAM use among Turkish asthmatic children was found to be high (66 %), with herbal medicine being the most popular modality. The results of this study shows that CAM use becomes more prevelant among asthmatic children from families with low income. It will be increasingly important for physicians who care for allergic children to be aware of high CAM usage.

Human Wharton’s Jelly-derived Mesenchymal Stem Cells (hWJ-MSCs) are easily available cells without transplant rejection problems or ethical concerns compared to bone-marrow-derived MSCs for prospective clinical applications. These cells display immunosuppressive properties and may be able to play an important role in autoimmune disorders. Regulatory T-cells (Treg) are important to prevent autoimmune disease development. Interleukin 35 (IL-35) induces the proliferation of Treg cell populations and reduces the activity of T helper 17 (Th17) and T helper 1 (Th1) cells, which play a central role in initiation of inflammation and autoimmune disease.Recent studies identified IL-35 as a new inhibitory cytokine required for the suppressive function of Treg cells. We created IL-35-producing hWJ-MSCs as a good vehicle for reduction of inflammation and   autoimmune   diseases.   We   isolated   hWJ-MSCs   based   on   explant   culture.   HWJ-MSCs were transduced at MOI=50 (Multiplicity of Infection) with lentiviral particles harboring murine Interleukin 35 (mIL-35). Expression of IL-35 in hWJ-MSCs was quantified by an IL-35 ELISA kit.IL-35 bioactivity was analyzed by inhibiting the proliferation of mouse splenocytes using CFSE cell proliferation kit. Frequency of CD4+CD25+CD127low/neg Foxp3+ Treg cells was measured by flow cytometry. There was an up to 85% GFP positive transduction rate, and the cells successfully released a high level of mIL-35 protein (750 ng/ml). IL-35 managed to inhibit CD4+ T cell proliferation with PHA, and improved the frequency of Treg cells.Our data suggest that transduced hWJ-MSCs overexpressing IL-35 may provide a useful approach for basic research on gene therapy for autoimmune disorders.

Individual preparation of two human T-cell lymphotropic virus type I (HTLV-I) diagnostic GST fused peptides (MTA-1 and GD21) is time-consuming and expensive. The aim of this study was to design a novel single chimeric antigen (SCA) to obviate separate expression of proteins and reduce the cost of reagent preparation.Structural protein fragments, including immunodominant B cell linear epitopes, were selected and different SCAs were designed. Tertiary structure, epitope exposure, solubility and stability were calculated for each SCA and compared with each other. The synthetic DNA  encoding  the  interested  SCA  was  sub-cloned  into  pET32a  expression  vector, expressed as a soluble form in Escherichia coli BL21 (DE3) cells and purified under native condition using affinity chromatography.The SDS-PAGE results indicated that thioredoxin-fused SCA was successfully expressed as a soluble form in E. coli BL21 (DE3) cells. The results of ELISA confirmed that SCA reacted with anti-HTLV-I antibodies in a concentration-dependent manner.Our results indicated that the designed SCA may be a good candidate for the screening of HTLV-I carriers with antigen–antibody-based tests.

The rs2476601 (R620W, C1858T) polymorphism in PTPN22 gene has been repeatedly reported to be associated with rheumatoid arthritis (RA). The rs 2476601 is widely suggested for predictive testing and risk assessment for RA. The aim of this study was to test the possible association of this SNP with RA in Iranian population.A total of 872 samples (405 confirmed RA patients and 467 healthy controls) were recruited in  this  study. Genomic  DNA  was extracted  from  whole  blood  and  the  genotyping  was performed  by  polymerase  chain  reaction–restriction  fragment  length  polymorphism  (PCR- RFLP). Genotyping for a set of samples were re-confirmed by two other rounds of genotyping, using another PCR-RFLP experiment with different enzyme and DNA sequencing.All 872 samples were genotyped as homozygous CC in first round of genotyping. Genotyping was repeated for 30% of samples by another restriction enzyme and for 10% of samples by sequencing. Again all samples showed homozygous CC genotype.This study suggests that the rs2476601 polymorphism of PTPN22 gene is mono-morphic in Iranian population, containing only C allele.Considering that previous studies in other populations reported the T allele as the risk allele at this locus, the present study concluded that rs2476601 play no role in susceptibility to RA and other autoimmune diseases in Iranian population. This finding has significant future clinical implications in determining the strategy for risk assessment and predictive testing for such diseases in Iranian population.

Interleukin-8 (IL-8) is a well-known inflammatory chemokine and suggested to be involved in the development of acne vulgaris. This study investigates IL-8 plasma levels in acne patients and healthy controls and the molecular basis for the regulation of the IL-8 gene in a Pakistani population.Patients with acne vulgaris (n=264) and healthy individuals (n=264) were enrolled in this investigation. Plasma IL-8 levels were determined by enzyme-linked immunosorbent assay (ELISA). The genotyping for IL-8 gene was performed by polymerase chain reaction (PCR) and  restriction fragment length polymorphism (RFLP).Our data showed a statistically significant increase in IL-8 levels from acne patients compared with healthy subjects (154.2 ± 52.1 pg/mL in patients vs. 101.6 ± 33.5 pg/mL in controls, p<0.0001). The IL-8-251T>A (rs4073) polymorphism was significantly higher in patients with acne compared with the control group (p=0.013). There was a significant difference between the T and A alleles from acne cases and controls (odds ratio OR=1.6, 95%CI=1.16-2.19,  p=0.003).  Logistic-regression  analysis  showed  that  the  increased IL-8  levels,  and  the  IL-8-251T>A  polymorphism  were  significantly  associated  with acne.Our data suggest that the elevated IL-8 levels and the IL-8-251T>A polymorphism may be associated with acne vulgaris in the study population.

We aimed to investigate IgG antibody levels specific to Toxocara canis (T. canis), a parasite which subsists in dog’s intestine, on serum samples obtained from patients with chronic urticaria (CU) to evaluate effective risk in CU etiopathogenesis.In this study, 73 patients diagnosed with CU and 109 healthy individuals as control group, were included. Various factors such as sex, age, education and income, daily hand washing habits, history of dog owning and soil eating were questioned in patient anamnesis. T. canis IgG antibodies were detected using an enzyme linked immunosorbent assay (ELISA) kit prepared with T. canis larval excretory-secretory antigens. Positive results were confirmed with western blot (WB) WB test.We found T. canis IgG positivity in 17.8% (n=13) of patients (n=73) with CU. But we did not observe any T. canis IgG positivity in healthy controls (n=109). Low molecular weight bands (24-35 kDa) were observed in 11 samples in WB analyses while two of the samples were weakly positive. It is revealed that dog owning history increases T. canis seropositivity12.9 times while insufficient daily hand washing habit (less than six times a day) increases seropositivity 20.7 times. Our study showed that T. canis may trigger CU since we found 17.8% seropositivity in 73 patients with CU and none in 109 healthy individuals.Moreover, various socio-demographic characteristics have been shown to affect T. canis seropositivity in patients with CU.

Severe combined immunodeficiency (SCID) represents a rare group of primary immunodeficiency disorders (PIDs), with known or unknown genetic alterations. Here, we report a new interleukin 2 receptor, gamma chain (IL-2RG) mutation in an Iranian SCID newborn.The patient was a 6-day old boy with a family history of PID. The child was screened using a molecular-based analysis for the assessment of T cell receptor excision circles (TRECs) and kappa-deleting recombination excision circles (KRECs). Moreover, a complete immunological evaluation and gene sequencing was performed.Results showed undetectable TREC but a high level of KREC copy numbers. Flow cytometric data indicated low numbers of T and NK cells, but elevated number of B cells. A novel substitution in IL2RG: c.675 C>A, leading to p.225 Ser>Arg was found. Based on the functional analysis, the mutation is predicted to be damaging. The patient was diagnosed as a T B+ NK X-linked SCID.