The major histocompatibility complex (MHC) genes are the most polymorphic loci in the human genome and have been widely studied in various populations and ethnic groups. Investigations into the HLA genes and proteins have been useful tool for anthropological, transplantation and disease association studies. The polymorphism of the HLA class I (A, B, C) and class II (DRB1, DQA1, DQB1) genes were investigated in 90 unrelated Iranian individuals from Yazd province located in the center of Iran using sequence-specific primers (PCR-SSP). Allele and haplotype frequencies, expected/observed heterozygosity, unbiased expected heterozygosity, number of effective alleles, deviations from Hardy-Weinberg (HW) equilibrium and genetic diversity were computed. A total of 23, 48, 23, 24, 13 and 16 alleles for HLA-A, -B,-C, -DRB1, -DQA and -DQB loci were determined, respectively in the population study. The most frequent allele identified in this study were A*02:01 (18.889%), HLA-B* 51:01 (12.778%), HLA-C* 12:03 (17.033%), HLA-DRB* 11 (24.4%), HLA-DQA* 05:05 (20.55%) and HLA-DQB*03:01 (22.8%).Furthermore, the most frequent 3-locus haplotypes were DRB*11-DQA*05:01-DQB*03:01 (21.1%), HLA-A*02:01- B *50:01- DRB*07:01 (4.9%) and A*26:01 –B* 38:01 –C*12:03(5%). The most 4-locus haplotype were A*11:01 –B* 52:01 –C*12:03 –DRB!*15(2.5%) and A*02:01 –B* 50:01 –DRB1*07:01 –DQB1*02:01(4.5%). The heterozygosity of the study population was confirmed the expected value and not deviated from Hardy-Weinberg equilibrium for all loci (p>0.05). Our study shows a close relatedness between Yazd population and other ethnic group of Iran despite some differences, which may be due to admixture of each one of these groups with each other or foreigner subpopulations during centuries. Moreover, the results of this study suggest that the Iranian population from Yazd province is in close vicinity with the Caucasians populations and far from the Korean and Japanese populations. 

There are confirmed beneficiary effects of exercise on atherosclerotic inflammation of ischemia-associated heart diseases. The purpose of this study was to evaluate the effect of aerobic exercise on T-regulatory cell markers of IL-35 as well as FoxP3 and T-helper2 marker of IL-33 in patients with ischemic heart disease (IHD). This research was performed on 44 asymptomatic male patients with ischemic heart disease. The participants were randomly assigned into two groups of submaximal aerobic exercise and control group. Blood samples were collected before and after the termination of the exercise protocol. Serum levels of IL-35 and IL-33 as well as the amount of FoxP3 gene expression in peripheral blood mononuclear cells were measured by Elisa and Real time PCR, respectively. Serum levels of IL-35 (p=0.001) as well as the amount of FoxP3 gene expression increased significantly (p=0.012)  in exercise group even after controlling the likely confounding effects of age, length of ischemia, duration of the disease, and the amount of such factors before exercise (p≤0.042). It seems that exercise may yield a better control of atherosclerotic inflammation in patients with ischemic heart disease through the induction of regulatory T cells. 

Multiple sclerosis (MS) is an inflammatory, multifocal, immune-mediated disease of the central nervous system that women are at a higher risk to acquire than men. Myxovirus resistance protein A (MxA) is used as a predictive marker of bioactivity of interferon-beta (IFN-β) therapy in MS patients. This study was undertaken in west of Iran to investigate gender differences in the expression level of MxA in relapsing-remitting MS (RRMS) patients receiving IFN-β therapy, compared with untreated normal individuals. The expression level of the MxA gene in RRMS samples were compared to untreated normal individuals using the extracted RNA from whole blood of 50 RRMS patients (31 females and 19 males) and 50 normal controls (29 females and 21 males). All patients were HLA-DRB1*15 negative and responded to IFN-β with a normal vitamin D level. The level of MxA gene expression was measured by quantitative RT-PCR. The levels of gene expression were decreased in RRMS patients compared with normal counterparts (p=0.025). This decrease was significant in females (p=0.009) compared to males (p>0.05). The level of expression varied across different female age-groups with no significant difference in women younger than 30 years, but a significant decrease in expression in women between 30 to 40 years or above 40 years of age was seen. There was neither linear correlation between the MxA expression level and risk of expanded disability status scale of Kurtzke (EDSS); nor were there any significant correlation between expression status of MxA and duration of the disease. In conclusion, the decrease in the level of MxA expression in MS patients treated with IFN-β when compared to normal individuals was significantly lower in females than males.  This demonstrated a gender bias in the response to IFN-β therapy that will need to be confirmed and further investigated in more detail.

The inhibition of the inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2) and nuclear factor-κB (NF-κB) production are research targets of attract in the field of anti-inflammatory drug development. Therefore, this study was designed to investigate the anti-inflammatory effects of novel thiazolidinone derivatives using a cellular model of lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7. In the present study, five new derivatives (A to E) of thiazolidinone were synthesized and screened for anti-inflammatory activities. Cell viability of LPS-stimulated RAW 264.7 macrophages clearly decreased in >55 μg/mL of synthesized A-E compounds especially in the presence of C; therefore, up to 50 μg/mL of compounds were selected for the subsequent analysis. A majority of these compounds showed significant inhibition on the production of NO in LPS-stimulated macrophages in a dose-dependent manner. Compounds B and D (10-50 μg/mL) significantly inhibited LPS-induced NF-κB (p65) production in a dose-dependent manner. The effects of B and D on iNOS and COX-2 mRNA and protein expression in LPS-stimulated RAW 264.7 cells were detected by real time-PCR and western blot. B derivative significantly suppressed the iNOS and COX-2 mRNA level and as well as protein expression. Taken together, these results reveal that compound B as new thiazolidinone derivative decreased expression of the inflammatory-related signals (NO, iNOS and COX-2) through regulation of NF-κB; hence, this compound could be suggested as a novel therapeutic strategy for inflammation-associated disorders.

Serum IgE assay is a mainstay step in the allergy work up. Allergenic extracts and molecular components are available at present. This real life study compared the serum specific IgE levels against allergenic extracts with allergenic molecules in patients allergic to Parietaria, Betulaceae, and mites. This retrospective real life study included 489 subjects with respiratory allergy. Inclusion criteria were 1) documented diagnosis of allergic rhinitis (AR) and/or allergic asthma, and 2) documented allergy to Parietaria judaica (Par j) 2 (216 patients: 112 females, mean age 42 years), or to Betula verrucosa (Bet v) 1 (62 patients: 35 females, mean age 3 years), or Dermatophagoides pteronyssinus (Der p) 1 (211 patients: 107 females, mean age 34 years); and mono-allergy. Serum IgE, specific both for total/crude allergen extracts and individual purified/recombinant allergens, were assessed by ImmunoCap system. The serum IgE levels to birch extract were very strongly (R²=0.96) related to IgE to Bet v 1. There was a strong (R²=0.71) correlation between Dermatophagoides pteronyssinus IgE and Der p 1. A very strong (R²=0.87) correlation also existed between Parietaria extract IgE and Par j 2 IgE levels. However, there was discrepancy between percentages of positivity between allergenic extracts and molecules. Therefore, allergen molecular diagnostics may represent a useful way in allergy work up, but deserves caution in particular circumstances.

Cow’s milk allergy (CMA) is the most frequent food allergy in children and oral immunotherapy (OIT) is a promising approach for treatment of patients. The most challenging cases are anaphylactic with coexisting asthma and proposing safe protocols is crucial especially in high risk groups. Considering that CMA varies among patients, an individualized OIT protocol would be beneficial to achieve a safer and more efficient method of desensitization. 18 children more than 3 years of age with IgE-mediated CMA were enrolled. CMA was confirmed by positive skin prick test (SPT) and positive oral food challenge (OFC) and 60% of individuals had a convincing history of persistent asthma. SPT with milk extracts, whole fresh milk and serially diluted milk concentrations were performed.  The dilution of milk that induced 3-5 mm of wheal in each individual was selected as the starting dilution for OIT. Desensitization began by 1 drop of the defined dilution and continued increasingly. Overall, 16 out of 18 children (88.8%) achieved the daily intake of 120 mL of milk. Four out of these 16 children accomplished the protocol without any adverse allergic reactions. 12 patients experienced mild to severe reactions. Wheal and erythema in SPT (p≤0.001), and sIgE (p≤0.003) to most milk allergens were significantly decreased following desensitization. We successfully desensitized 16 of 18 children with IgE-mediated CMA by individualized desensitization protocol. Individualizing the OIT protocol would be helpful to save time and perhaps to relieve the allergic symptoms after ingesting cow’s milk intake.

Fractional exhaled nitric oxide (FeNO) has been suggested as a non-invasive biomarker of airway inflammation, which is increased in atopic subjects. Whether sensitization to particular allergens is a predictive factor for increased FeNO levels is not yet fully understood. We conducted a retrospective cross-sectional study. From October to December in 2015, the medical documents of 127 mild, steroid-naive asthmatic children and 34 healthy age-matched children were enrolled in this study. The results of the FeNO measurements, skin prick test, and the spirometry were collected for analysis. Sensitization patterns to the 18 aeroallergens (5 categories: mites, molds, animal dander, pollen, and other) were determined in study population. A significant increase in FeNO level was observed in poly-sensitized asthmatic children (34.7 part per billion, (ppb) [28.3-41.1 p.p.b]), compared with mono-sensitized asthmatics (30.7 p.p.b [18.3-43.2 p.p.b]) and with non-sensitized asthmatics (17.3 p.p.b [10.8-24.5 p.p.b]). With sensitization to perennial allergens (mites, mold, and animal dander), blood eosinophil counts were significantly associated with increased FeNO (p<0.05 for all). The highest FeNO level was identified in children sensitized to a combination of the perennial, seasonal, and other allergens, when compared with those sensitized to one category of allergen alone (p=0.004). Our study showed that variations in FeNO level were associated with individuals’ sensitization patterns. Being sensitized to some particular allergens might contribute to prompt the airway inflammation. 

There are many studies related to the production of a ELISA kit for diagnosing virus infections. However, production of most kits depends on purification of whole virus particles, which involves the use of costly equipment and reagents. The purpose of this study was to check out if the anti-CP42 antibodies could be used as a diagnostic assay for detection of Grapevine fanleaf Virus (GFLV). In this study, recombinant GFLV coat protein gene related to selected antigenic determinants was inserted into pET-28a bacterial expression vector and the construct (pET-28a CP42) was cloned into E. coli strain (DE3). Expressed protein was verified with western blotting assay by the use of commercially available anti-GFLV antibody. The recombinant protein was purified using nickel–nitrilotriacetic acid (Ni–NTA) resin. Balb/c mice were immunized with purified protein and splenocytes of hyperimmunized mice were fused with murine myeloma Sp2/0 cells. Positive hybridomas were selected by ELISA using CP42 as coating antigen. The results showed that monoclonal antibody (MAb) specific to CP42 has been successfully generated. Efficiency of produced antibody was analyzed by ELISA and western blotting assay using some confirmed grapevine samples. The infection was confirmed previously based on morphological features and ELISA assay, performed using commercial anti-GFLV antibody. The monoclonal antibody reacted with antigen in ELISA and immunoblot method. Our results demonstrated that anti recombinant CP42 monoclonal antibodies are able to diagnose whole virus in infected grapevine sample using ELISA test.

Etiologic factors for recurrent miscarriage (RM) include autoimmune diseases, the most frequently antiphospholipid syndrome and thyroiditis. Some women who suffer from RM might also have an altered immune system. We aimed to evaluate possible associations between anti-thyroid and anti-phospholipid antibodies in women with RM. In a retrospective case series 1 on 156 women with RM, major outcome parameters were antibodies against cardiolipin, β2-glycoprotein I, thyreoperoxidase (TPO-Ab), and thyroglobulin (TG-Ab). Significant (p<0.05) positive correlations were found between TPO-Ab and TG-Ab (r=0.577), TPO-Ab and IgG anti-cardiolipin antibodies (r=0.284), TPO-Ab and IgG anti- β2-glycoprotein I antibodies (r=0.196), and TG-Ab and IgG anti-cardiolipin antibodies (r=0.193), as well as between all types of anti-phospholipid antibodies. Women with both increased TPO-Ab and TG-Ab levels revealed higher (p<0.001) IgG anti-cardiolipin and IgG anti-β2-glycoprotein I antibodies. Anti-thyroid antibodies were linked to anti-phospholipid antibodies and should be in the focus of future research on RM.

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