Siglec-F (SF) is a surface glycoprotein expressed by mouse eosinophils and induces caspase- and mitochondria-dependent apoptosis after engagement with its cognate ligand or specific antibodies. This targeting eosinophils by monoclonal antibodies may help diverse diseases associated with increased frequency of eosinophils including allergy and asthma. In this paper, production of murine and rat monoclonal antibodies (mAbs) against Siglec-F has been addressed. Balb/c mice were immunized with siglec-F1 (SF1) and siglec-F2 (SF2) synthetic peptides conjugated to a carrier protein. Rats were immunized with Chinese hamster ovary CHO cells overexpressing Siglec-F (CHO-SF) or with Siglec-F-human immunoglobulin FC fusion protein (CHO-SF-Ig). Hybridomas were produced by standard protocol and screened for their reactivity by enzyme-linked immunosorbent assay (ELISA), western blotting (WB), and flow cytometry. In parallel, polyclonal antibodies were generated in New Zealand White rabbits immunized with SF1 and SF2 peptides. Three mouse and three rat mAbs were generated against synthetic peptides and SF-Ig, respectively. All mouse monoclonal and rabbit polyclonal antibodies reacted well with immunizing molecules in ELISA and detected specific band of Siglec-F in WB. However, they failed to detect native molecule in flow cytometry analysis. Quite the contrary, rat mAbs did not reacted with the denatured protein in WB, instead exhibited significant reactivity with CHO-SF cells in flow cytometry. Based on the heavily glycosylated nature of Siglec-F, it seems that generation of anti-SF antibodies able to detect native protein needs a properly folded molecule for immunization. Monoclonal antibodies reported here are invaluable tools for studying linear and conformation epitopes of SF and tracing mouse eosinophils.

CD247 and CD226 play important roles in signaling of lymphocytes. Single nucleotide polymorphisms (SNPs) of genes encoding CD247 and CD226 have been associated with the risk of several autoimmune disorders. This study aimed to evaluate the possible association between CD226 and CD247 genes SNPs and risk of systemic sclerosis (SSc) in Iranian population. Study participants were 455 SSc patients and 455 age, sex and ethnic -matched healthy individuals. Genotyping of rs2056626 and rs763361 at CD247 and CD226 genes, respectively, was carried out using TaqMan MGB-based allelic discrimination real-time PCR. Neither alleles nor genotypes of both SNPs showed significant association with the risk of SSc. Furthermore, association analysis of the genotypes with clinical manifestations of the disease revealed that rs763361 variants were associated with the forced vital capacity (FVC) in SSc patients. Our results suggest that genetic variants of CD226 and CD247 genes may not be a contributing factor in pathogenesis of SSc in Iranian population.

Despite the increasing prevalence of anaphylaxis, there is little information about the characteristics and practice of healthcare providers in treating anaphylaxis, so this study was conducted to record the characteristics and therapeutic approaches of anaphylaxis from May 2012 until April 2015, the data of all patients diagnosed with anaphylaxis in the Allergy department of three referral university hospitals in Tehran, Iran were recorded. Thereafter, the demographics, clinical features, triggers and therapeutic approach were evaluated. This study investigated 136 individuals, 64 males (47%) between 6 months and 68 years old, as well as 72 others (52.94%) under 18 years of age (pediatric). The following were the most common organs involved: Skin 86.02% (pediatric 91.66% vs adult 79.68%), respiratory tract 51.47% (pediatric 43.05% vs adult 60.93%), cardiovascular 50.73% (pediatric 54.16% vs adult 46.87%), gastrointestinal 20.58% (pediatric 27.7% vs adult 12.5% ) and neurologic system 5.88% (only in adults). The following were the most identified causing foods 69 (50.37%)[42 pediatric (children) and 27 adults], drugs 34( 25%)[14 pediatric and 20 adults], idiopathic 16( 11.77%)[3 pediatric and 13 adults], insect sting 7( 5.15%)[3 pediatric and 4 adults] , exercise 6( 4.42%) [1 pediatric and 5 adults]. Milk, egg and wheat were the most common causative foods in pediatric cases but sesame, as well as egg and milk were the most common causes in adults. Epinephrine injection, auto injector epinephrine prescription as a discharging plan and referral to an allergist were: 10.78, 1.96 and 7.8 %, respectively. In this case series we found that, cutaneous, respiratory, cardiovascular and gastrointestinal complains were the most common manifestations and food, drug and idiopathic were the most common causes.In this study, the diagnosis of anaphylaxis, epinephrine subscription and referral to an allergist were significantly lower in comparison to other studies.

The aim of this study was to evaluate the associations between caregiver-reported use of medications, alcohol, cigarette and/or waterpipe (WP), and exposure to pesticides/detergents during pregnancy with childhood-onset asthma. The study design consisted of a case-control study, conducted between December 2015 and April 2016, recruited 1503 children, aged between 3-16 years old. A questionnaire assessed the sociodemographic characteristics (age, gender, education level of both parents), the family history of asthma, and other known risk factors of asthma (heating system at home, child history of recurrent otitis, humidity in the house, child went to a daycare, smoking and drinking alcohol during pregnancy, exposure to pesticides and detergents). The multivariate analysis showed that children living in North and South Lebanon and the children living in areas where pesticides are frequently used had an increased risk of asthma (ORa=1.625, CI 1.034-2.554, p=0.035, ORa=13.65, CI 3.698-50.385; p<0.001 and ORa=3.307, CI 1.848-5.918, p<0.001 respectively). Smoking WP during pregnancy and cigarette during lactation would increase the risk of asthma in children (ORa=6.11; CI 1.244-30.008; p=0.026 and ORa=3.44; CI 1.024-11.554; p=0.046 respectively). We conclude that asthma may originate from the environmental exposure to toxics such as pesticides and tobacco (cigarettes and WP) or to alcohol and prescribed medications during pregnancy and lactation. Spreading awareness by health professionals about these preventable causes can help educate the parents and children to prevent asthma and its exacerbation. 

In recent studies, mesenchymal stromal cells (MSCs) have been increasingly employed to treat various diseases like pulmonary fibrosis (PF). There are very few MSCs in tissues so in order to obtain their sufficient numbers for therapeutic applications, their in vitro expansion is necessary. The aim of this study was to investigate the effects of long-term culture of the human umbilical cord vein MSCs (hUCV-MSCs) on pulmonary fibrosis in mice. MSCs were first isolated from human umbilical cord vein and cultured up to 18 passages. In C57BL/6 mice, 15 min after belomycin instillation, UCV-MSCs at passages (P) 0, 4, 8, 12, and 18 (long-term culture) were transplanted intratracheally. Mice were weighted every 5 days and were euthanized on day 21. For histopathological examination, the lung sections were stained with hematoxylin-eosin (HE) and Masson’s trichrome. The mRNA expression of TGF-β1, alpha-smooth muscle actin (α-SMA), and collagen type I alpha 1 (COL1A1) in lung tissues were assessed using RT-PCR. For cell tracking, human cytochrome B DNA was detected in mice lung tissues by PCR. The weight of mice receiving long-term culture of UCV-MSCs increased compared to other mice (p=0.056). Also, transplantation of UCV-MSCs at P18 led to increased alveolar space and decreased connective tissue and collagen deposition of the lung tissues. The mRNA expression of TGF-β1, α-SMA, and COL1A1 also decreased in this group. The results showed that intratracheally transplanted long-term culture of the UCV-MSCs attenuated lung fibrosis in mice. 

Recent studies suggest that imbalances in the ratios of CD4+ T helper cell subsets, T helper-17 (Th17) and regulatory T (Treg) cells play a crucial role in the pathogenesis of acute lung injury (ALI). However, studies of the imbalance of Th17/Treg in paraquat (PQ)-induced ALI have not been reported. Therefore, we investigated whether the ratio of Th17/Treg cells in a mouse model of PQ-induced ALI contributes to pathogenesis of ALI. Male Kunming mice were randomly treated with saline (control group) or PQ (PQ-poisoned (PQP) group); mice were sacrificed at either 12 hours (PQP-12h) or 24 hours (PQP-24h and control) post-treatment. Hematoxylin-eosin and TUNEL staining procedures were performed to examine inflammation and apoptosis. The presence of Th17 and Treg cells was measured by flow cytometry; the expression of putative Th17 cytokines and transcription factors was measured by ELISA and western blot analysis. Compared with control mice, lung inflammation and apoptosis were dramatically increased in PQP mice at 12 and 24 hours after poisoning. In addition, poisoned mice displayed significant increases in the presence of CD4+IL-17+ T cells (Th17) and in the expression of IL-17A and IL-17, as measured by flow cytometry and western blot assays. This increase was most notable after 24 hours of PQ exposure. Furthermore, poisoned mice displayed marked decreases in the presence of CD4+CD25+Foxp3+ T cells (Treg) and in the expression of IL-35 and the transcription factor Foxp3. These results suggest that an imbalanced ratio of Th17/Treg cells may contribute to the pathogenesis of PQ-induced ALI.

Serous otitis media with effusion (OME) is a middle ear inflammatory response to allergens and microbes which stimulate leukocytes to produce different inflammatory mediators after obstruction of Eustachian tube. Here, we investigated the levels of these mediators, IL-17 and IL-23, in serum and middle ear fluids of children with OME. 75 patients with otitis media and 75 age and sex-matched healthy controls were enrolled in this study. IL-17 and IL-23 levels in serous secretion of the patients and their serum levels were measured in both groups by ELISA. Serum IL-17 levels were significantly higher in the patients than controls (p=0.001). There was no significant difference between serum IL-23 levels in patients and controls. Patients’ serous levels of both cytokines of IL-17 and IL-23 were higher than those in serum according to different parameters of sex, age, and duration of the disease. This study shows an elevated presence of IL-17 and IL-23, as pro inflammatory cytokines, in OME. These finding may represent the contribution of such cytokines in the pathogenesis of OME. Blocking such molecules may yield new non-surgical therapeutics. 

Asthma is a chronic inflammation of the airways affecting over 300 million people worldwide. As in the autoimmune diseases, it is well described that women are the most affected by asthma. The higher number of women presenting this pathology suggests the involvement of female sex hormones in the construction of the allergic immune response. Female Balb / c mice were used for the experiments. Thirty-eight animals were separated into four groups: OVX-Ova; Sham-Ova; OVX-Sal; Sham-Sal. Then animals underwent acute allergic induction protocol by Ovalbulmin (OVA). Ovariectomized animals showed greater number of leukocytes in bronchoalveolar lavage (BAL) and elevated white blood cells recruitment to the lung environment observed by histological analysis. There was a significant increase of eosinophils and mast cells in inflammatory sites at pulmonary tissue. The relative uterine and body weight were lower in ovariectomized animals and higher in Sham mice, respectively. Moreover, the lack of the sex hormones induced an increase in interleukin (IL)-4 and titers of immunoglobulin G1 (IgG1) antibodies. However, increased production of IL-17A was only observed in Sham animals. Altogether, data this study suggest that ovariectomy induces the formation of a stronger Th₂ response in allergic animal. However, the immune processes involved in the allergic response in females currently remain unclear.

The liver has unique microenvironment which is known to induce tolerance of cytolytic CD8⁺ T cells to hepatic and extra hepatic antigens, resulting in persistence of infection of the liver by the hepatitis B and C viruses. However, under some conditions, functional immune responses can be elicited in the liver in particular to show preferential retention of activated CD8⁺ T cells. It is not clear whether this retention depends on the type of the exogenous immunostimulatory or the endogenous innate immune cells. The T cell receptor (TCR) transgenic OT-1 (CD8⁺) mouse model was used in which OT-1 cells were harvested from the spleen of the donor and transferred into recipient mice followed by immunization with OVA peptide followed by injection of GM-CSF, CCL21 chemokine, or cytokines (IL-2, IL-12, or IL-15), or the toll-like receptor 3 agonist poly(I:C). Co-administration of any of these immunostimulatory agents relatively augmented the retention of CD8⁺ T cells with different levels of effects. Compared to spleen, the Ag-specific CD8⁺ T cells in the liver showed higher activities including expansion, proliferation, apoptosis and memory responses as well as cytolytic function. While depletion of natural killer cells significantly decreased the hepatic retention of the antigen-specific T cells, depletion of Kupffer cells showed opposite effect. Taken together, the antigen reactive T cells in the liver have higher activities than their counterparts in the peripheral tissues such as spleen. These data have important clinical implications for designing immunotherapeutic protocols toward the liver diseases.

The present study aimed to compare the levels of high-mobility group box 1(HMGB1) and soluble triggering receptor expressed on myeloid cells (sTREM1) in the gingival crevicular fluid (GCF). This cross-sectional cohort trial investigated two groups of 22 eligible chronic periodontitis and 22 periodontally healthy individuals (student volunteers) both before and after the periodontal treatment.  GCF was collected from the deepest pockets with clinical attachment loss≥3 mm. Both groups received oral hygiene instructions, and scaling and root planning were performed in the test group. Enzyme-linked immunosorbent assay kit (ELISA) was used to measure the levels of HMGB1 and sTREM1 in GCF samples collected before and 1 month after non-surgical periodontal treatment. The results showed that HMGB1 levels were significantly higher in the chronic periodontitis patients than those of the healthy individuals before treatment (p<0.02) and decreased significantly after periodontal treatment, which reduced gingival inflammation. Furthermore, the levels of sTREM1 marker were significantly higher in periodontitis patients before (p<0.001) and 1 month after treatment than in healthy individuals (p<0.003) although its crevicular levels decreased after periodontal therapy in periodontitis group. The higher levels of sTREM1 and HMGB1 cytokines in GCF of periodontitis patients and the significant decrease after the introduction of the periodontal treatment underlines the importance of HMGB1 and sTREM1 in pathogenesis of periodontitis.

Eosinophilic granulomatosis with polyangiitis is a systemic vasculitis. It could affect respiratory system, kidney, and central nervous system frequently; however, all body organs could be involved. Asthma and eosinophilic pneumonia are predominant manifestations in respiratory system. Bronchoalveolar lavage or lung biopsy may be used for diagnosis, but endobronchial lesion is not considered as a manifestation of eosinophilic granulomatosis with polyangiitis. Here we present a case of eosinophilic granulomatosis with polyangiitis with unusual endobronchial lesion which was confirmed by endobronchial biopsy.

An acute coronary syndrome (ACS) occurring during the course of an allergic reaction is called Kounis syndrome (KS). The second case of KS induced by diclofenac potassium (DP) is presented in this report. A 67-year-old man was brought to our emergency department with the possible diagnosis of anaphylactic shock by the ambulance staff. It emerged that widespread erythema and pruritus developed after taking DP. Then, he lost consciousness. Diffuse urticarial lesions were detected on physical examination at the emergency department. He complained of chest pain during his observation, and progressive ST segment elevation was seen in the inferior leads on serial electrocardiograms. His coronary angiography showed 100% occlusion of the right coronary artery.  Then, KS was diagnosed. The patient was discharged on the second day, and he was doing well on the control visit 2 weeks later. All allergic reactions may trigger an ACS so physicians should be aware of KS and always keep that unique clinical entity in mind to recognize it promptly and direct the therapy at suppressing the allergic reaction and improving the coronary circulation simultaneously when encountering a patient with symptoms suggesting an allergic reaction and a concomitant ACS.