Evaluation of CD137 and CD137L Transcript Levels and the Serum sCD137 in Immune-mediated Polyneuropathy.

Background: Abnormal humoral and cellular immune responses have been reported in immune-mediated polyneuropathies. CD137, as a costimulatory molecule and a TNF receptor superfamily member, has been demonstrated to have a key role in the pathogenesis of many autoimmune as well as inflammatory disorders. Objective: To evaluate the transcripts levels of CD137, its ligand (CD137L), and the serum levels of soluble CD137 (sCD137) in patients with immune-mediated polyneuropathy. Methods: A total of 45 patients and 46 sex and age-matched healthy individuals were enrolled in the study. CD137 and CD137L transcript levels were assessed by the Real-Time PCR, and the serum level of sCD137 was measured using the ELISA technique. The Bayesian regression model was used for statistical analysis at the 0.05 significance level in R 4.1.0 statistical environment. Results: Transcript levels of the CD137 and CD137L were higher in polyneuropathy patients in comparison with the healthy subjects (P=0.006 for both). Conversely, the mean level of sCD137 was significantly lower in the sera of patients compared to the controls (P<0.001). Conclusion: Our findings point to the possible role of CD137 and CD137L in immune-mediated polyneuropathy pathogenesis. More investigations are required to clarify the exact contributions of the mentioned molecules to the pathogenesis of immune-mediated polyneuropathies.

Over-expression of IL-6 coding gene in the peripheral blood of migraine with aura patients.

Migraine is a common disorder which is placed among the top ten reasons of years lived with disability. Cytokines are among the molecules that contribute in the pathophysiology of migraine. In the current study, we evaluated expression levels of IL-6 coding gene in the peripheral blood of 120 migraine patients (54 migraine without aura and 66 migraine with aura patients) and 40 healthy subjects. No significant difference was detected in expression of IL-6 between total migraine patients and healthy controls (Posterior beta = 0.253, P value = 0.199). The interaction effect between gender and group was significant (Posterior beta =-1.274, P value = 0.011), therefore, we conducted subgroup analysis within gender group. Such analysis revealed that while expression of this gene is not different between male patients and male controls (Posterior beta =-0.371, P value > 0.999), it was significantly over-expressed in female patients compared with female controls (Posterior beta = 0.86, P= 0.002). Expression of IL-6 was significantly higher in patients with aura compared with controls (Posterior beta = 0.63, adjusted P value = 0.019). However, expression of this cytokine coding gene was not different between patients without aura and healthy subjects (Posterior beta = 0.193, adjusted P value = 0.281). Therefore, IL-6 might be involved in the pathophysiology of migraine among females and migraine with aura among both sexes.

Assessment of Apoptosis Pathway in Peripheral Blood of Autistic Patients.

Autism spectrum disorder (ASD) includes a number of severe neurodevelopmental disorders known by defects in social interaction, impaired verbal and non-verbal interactions, and stereotypic activities and limited interests. Dysregulation of apoptotic pathways have been demonstrated in brain tissues of affected individuals. In the present study, we evaluated expression levels of apoptosis-related genes and miRNAs in peripheral blood of ASD patients compared with healthy subjects. Transcript levels of BCL2, CASP8, and hsa-29c-3p were significantly lower in total ASD patients compared with total normal children (P values = 0.003, 0.002, and 0.01 respectively). When sex of study participants was considered in the analysis, the difference in transcript levels of these genes was significant only in male subjects. Peripheral expression of BCL2 and hsa-29c-3p had 100% sensitivity 92% specificity in ASD diagnosis. The diagnostic power of combination of transcript levels of these genes was estimated to be 78% based on the calculated AUC value. The present study provides evidences for dysregulation of apoptotic pathways in peripheral blood of ASD patients and suggests certain apoptosis-related genes as biomarkers in this regard.

MALAT1 Genomic Variants and Risk of Multiple Sclerosis.

Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is a long noncoding RNA (lncRNA) with a possible role in the regulation of immune responses. A previous study has demonstrated down-regulation of this lncRNA in multiple sclerosis (MS) patients. In the current study, we genotyped two MALAT1 single nucleotide polymorphisms (SNPs) in 428 Iranian MS patients and 505 healthy subjects. The G allele of the rs619586 was significantly under-represented in MS patients compared with controls (OR (95% CI) = 0.65 (0.46-0.92), adjusted P value = 0.03). This SNP was associated with lower MS risk in dominant model (OR (95% CI) = 0.63 (0.43-0.91), adjusted P value = 0.03). The rs3200401 was not associated with MS risk in any inheritance model. Moreover, the A T haplotype (rs619586 and rs3200401, respectively) within MALAT1 was associated with MS risk. The current study provides additional evidences for contribution of MALAT1 in the pathogenesis of MS.

Effect of Extremely Low Frequency Electromagnetic Fields on Expression of T-bet and GATA-3 Genes and Serum Interferon-γ and Interleukin-4.

This study investigated the effect of various magnetic flux densities of extremely low frequency electromagnetic fields (ELF-EMF) on expression of T-box transcription factor (T-bet) and GATA binding protein-3 (GATA-3) genes in the spleen and thymus of rats injected with human serum albumin (HSA). Moreover, serum levels of interferon (IFN)-γ and interleukin (IL)-4 were evaluated at two phases, that is, prestimulation and poststimulation with HSA. Eighty rats were separated into five groups, and four groups were exposed daily to 50 Hz EMF of 1, 100, 500, and 2000 µT magnetic flux densities for 60 days. To activate the immune system, 100 µg HSA was intraperitoneally injected into each rat on days 31, 44, and 58 of the regimen. Splenic and thymic T-bet and GATA-3 messenger RNA (mRNA) expression on day 61 was evaluated by reverse transcription quantitative PCR. Serum IFN-γ and IL-4 (in blood on day 31 before HSA and again on day 61) levels were evaluated by enzyme-linked immunosorbent assay. Expression of T-bet and GATA-3 mRNA was decreased in the spleen in hosts exposed to densities of 1 and 100 µT. Serum IFN-γ and IL-4 levels were also significantly decreased in 100 µT-exposed rats, but only at the prestimulation phase. From these findings, it appears that (30 and 60 days) ELF-EMF exposure could suppress the expression of some key genes associated with T helper (Th) cells and on some of their associated functions, that is, the ability to generate (in some cases, spontaneously) select cytokines. Whether this is attributable to effects on Th1/Th2 levels in the hosts and/or due to potential effects of the EMF on cellular functions remains to be determined.

Extremely Low Frequency Electromagnetic Fields Decrease Serum Levels of Interleukin-17, Transforming Growth Factor-ß and Downregulate Foxp3 Expression in the Spleen.

The study aimed to determine effect of extremely low frequency (50 Hz) electromagnetic fields (ELF-EMFs) exposure on serum levels of interleukin-17 (IL-17) and transforming growth factor-ß (TGF-ß) as signature cytokines of Th17 and regulatory T (Treg) cells, respectively. Retinoid-related orphan receptor γT and transcription factor forkhead box P3 (Foxp3) expression levels as lineage defining of Th17 and Treg cells were also assessed in the spleen and thymus. Eighty male rats were separated into 4 exposed groups (1, 100, 500, and 2,000 µT magnetic flux intensities) and a control. All rats were immunized by human serum albumin after 1 month of the exposure and the experiment was continued in the same manner for 1 month more. The results demonstrated that the weight of thymuses was significantly declined at intensity of 2,000 µT. At the preimmunization phase, the serum levels of IL-17 and TGF-ß were significantly decreased at intensities of 1 and 100 µT. The expression of Foxp3 was also downregulated at intensities of 1 and 100 µT. In conclusion, low intensities of ELF-EMF may reduce the serum levels of IL-17 and TGF-ß and downregulate the expression of Foxp3 in spleen.

Single-Nucleotide Polymorphisms in Interleukin 6 (IL-6) Gene Are Associated with Suicide Behavior in an Iranian Population.

Previous association studies have demonstrated the association between immune regulatory genes and suicide behavior. Among these genes are those coding for cytokines. In the present study, we genotyped two interleukin 6 (IL-6) variants (rs2069845 and rs1800795) in 320 suicide attempters, 236 suicide completers, and 341 individuals without any history of psychiatric disorders or suicide ideation. The rs2069845 was not associated with suicide behavior. The rs1800795 C allele was significantly more common among suicide completers compared with suicide attempters (OR (95% CI) = 1.33 (1.04-1.71)), adjusted P = 0.04). Besides, the rs1800795 was associated with the lethality of suicide attempt in recessive model (adjusted P value = 0.01). Consequently, the present study supports the role of IL-6 in suicide behavior and warrants further researches to confirm its effect and explain the underlying mechanism.

Cytokine profile in autistic patients.

The etiology of Autism Spectrum Disorders (ASDs) as severe neurodevelopmental ailments is not known. However, several evidences point to dysregulation of immune system as an underlying cause of ASD. In the present study we evaluated the mRNA expression levels of TNF-α, TGF-ß, IFN-γ, CXCL8, IL-1ß, IL-2, 1L-4, IL-6, IL-17 in whole blood samples of 30 ASD patients and 41 age and sex-matched healthy subjects with means of real-time PCR. TNF-α, IL-6 and IL-17 have been shown to be significantly up-regulated in ASD patients compared with healthy subjects (P < 0.0001, P = 0.001 and P < 0.0001 respectively). IL-2 has been shown to be significantly down-regulated in total ASD patients (P < 0.0001). No significant difference has been found in expression levels of other cytokines between patients and healthy subjects. The present study provides further evidences for dysregulation of immune response in ASD patients.

Expression Analysis of Long Non-coding RNAs in the Blood of Multiple Sclerosis Patients.

Multiple sclerosis (MS) is a chronic immune-mediated disorder of the central nervous system (CNS) with multiple genetic and environmental risk factors. Long non-coding RNAs (lncRNAs) have been recently reported to participate in the regulation of immune responses. Consequently, aberrant expression of lncRNAs has been suggested as an underlying cause of MS. In the present study, we evaluated the expression of three lncRNAs with putative roles in the regulation of immune response, namely TNF-α and heterogeneous nuclear ribonucleoprotein L (THRIL), Fas cell surface death receptor- antisense 1 (FAS-AS1), and plasmacytoma variant translocation 1 (PVT1) in circulating blood cells of 50 Iranian relapsing-remitting multiple sclerosis (RRMS) patients compared with healthy subjects by means of quantitative real-time polymerase chain reaction (PCR). We detected a significant downregulation of PVT1 and FAS-AS1 expressions in RRMS patients while a significant upregulation of THRIL in patients compared with controls (P < 0.001). Correlation analyses between lncRNA expression levels and clinical data of MS patients revealed no significant correlation between lncRNAs expression levels and Expanded Disability Status Scale (EDSS), a moderate correlation between PVT1 expression levels and duration of the disorder and no significant correlation between lncRNAs expression levels and age at onset. In addition, we demonstrated correlations between the expression levels of PVT1 and THRIL as well as expression levels of THRIL and FAS-AS1 in RRMS patients. In brief, we have demonstrated dysregulation of three lncRNAs in MS patients. Further studies are needed to explore the exact mechanisms by which these lncRNAs participate in regulation of immune responses.

The Importance of VEGF-KDR Signaling Pathway Genes should Not Be Ignored When the Risk of Developing Multiple Sclerosis is Taken into Consideration.

Vascular endothelial growth factor (VEGF) and its receptor kinase insert domain-containing receptor (KDR) pathway trigger the process of angiogenesis as well as inflammation, which contributes to the development and progression of demyelinating lesions in multiple sclerosis. This work is a case-control study comprising of a total of 400 subjects with multiple sclerosis and 400 healthy controls. Participants were subjected to neurological examination and peripheral blood sampling for genotyping. Polymorphisms in the VEGF and KDR genes were assessed using the restriction fragment length polymorphism (RFLP-PCR) method. A significantly higher frequency of the T allele and TT genotype of the VEGF 936C > T (rs3025039) polymorphism was found in the multiple sclerosis group than in the healthy control group (P = 0.01 [OR = 1.41] and P = 0.01 [OR = 3.12], respectively). In addition, VEGF 936C  >  T showed an association with patients in a recessive model. However, the KDR -604T > C (rs2071559) polymorphism showed no significant difference in either allelic or genotype frequency between the two groups. Taken together, the results of the present study suggests that the T allele of the rs3025039 in VEGF gene could be considered a risk factor for developing multiple sclerosis in the Iranian population.

Phospholipase D1 expression analysis in relapsing-remitting multiple sclerosis patients.

Multiple sclerosis (MS) is a chronic disorder resulting from destruction of the myelin or insulating covers of neurons in the central nervous system (CNS). Several lines of evidence suggest a role for immune response in the occurrence and progression of this disorder. Several disease-modifying agents (DMA) including ß-interferons (IFNß) are being used in MS patients in order to stop the disease at the early inflammatory stage, postpone disease progression and diminish future disability. Phospholipase D1 (PLD1) is a critical enzyme responsible for the making lipid second messenger phosphatidic acid. It has an established function in regulation of immune response. In the present study we have evaluated PLD1 transcript levels and plasma concentrations in 78 relapsing-remitting MS (RRMS) patients as well as 78 normal age- and sex-matched healthy subjects using real-time quantitative RT-PCR and enzyme-linked immunosorbent assay (ELISA), respectively. Significant PLD1 down-regulation has been observed in total MS patients compared with controls (P < 0.001) as well as IFN-ß responders (P = 0.034) and non-responders (P < 0.001) compared with controls, respectively. However, a significant up-regulation has been detected in IFN-ß responders compared with non-responders (P = 0.047). In both males and females groups, significant down-regulations have been detected in patients compared with controls (P = 0.014 and P = 0.002, respectively). The same results have been detected in PLD1 plasma concentrations. In conclusion, PLD1 transcripts in blood and its plasma concentrations can be used as putative biomarkers for evaluation of therapeutic responses to IFN-ß in RRMS patients. However, this result should be validated in future studies.

Evaluation of antibodies to cytomegalovirus and Epstein-Barr virus in patients with autism spectrum disorder.

Autism is a neurodevelopmental disease that manifested by a wide range of behavioral disorders. Although the etiology of autism is remained unknown but it is suggested that ASD have a complex etiology, including genetic and environmental factors, which may explain the observed different behavioral disorders in these patients. One of the proposed reasons for autism is viral infection in the early stages of development. The mechanism by which viral infection could lead to autism is still unclear.Previous studiesemphasized on the role of family membersof Herpesviruses in autism susceptibility. In this study, anti-Cytomegalovirus (CMV) and anti-Epstein-Barr virus (EBV) antibodies in the serum of 45 children with autism and 45 healthy individuals were evaluated. Serum samples were isolated from 5 ml blood of the patients and controls. Sandwich ELISA was used to quantitatively measure antibodies against the mentioned viruses. Results analyzed by SPSS software showed an increased amount of anti-CMV IgG and IgM antibodies in the blood of patients with Autism but not statistically significant (P< 0.05). The anti-EBV IgM antibody in the blood of patients with Autism was not only increased but also statistically significant (P< 0.05), however, the IgG level against EBV in the serum of ASD patients showed no significant difference in comparison to healthy controls. So it can be said that although the mechanisms of viral infection in autism is unknown, but probably EBV infection is associated with an increased risk of autism.

Frequency of viral infections and environmental factors in multiple sclerosis.

OBJECTIVES: Multiple sclerosis (MS) is a complicated disease which occurs due to relationship between genes and environmental factors that causes tissue damage by autoimmune mechanisms.We investigated and illustrated the hypotheses correlated to the evidence of several putative environmental risk factors for MS onset and progression in this part of Iran. MATERIALS AND METHODS: Univariate logistic regression was used to detect the effects of environmental factors on the risk of MS. Data were analyzed using SPSS version 16. RESULTS: The childhood history of patients with rubella, measles and chickenpox increased the risk of MS significantly. Moreover, low consumption of dairy products, avoidance of seafood consumption, cigarette smoking and exposure to tobacco smoke, stress, anxiety disorders, depress and disturbing thoughts, negative and disturbing thoughts, developing a sudden shock upon hearing bad news, having obsessive-compulsive and being depressed increased the risk of MS significantly. CONCLUSIONS: The results of the current research partially solved the puzzling question of complex interplay between environmental factors and MS disease in this part of Iran. Incorporating these factors enables more powerful and accurate detection of novel risk factors with diagnostic and prognostic methods.

TRAIL gene expression analysis in multiple sclerosis patients.

BACKGROUND: Multiple sclerosis (MS) as an autoimmune disorder in which the insulating covers of neurons in the Central Nervous System are destructed. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is an immunomodulatory molecule to protect against T cells hyper activation. METHODS: In this Case-control study, we compare TRAIL gene expression in peripheral blood between 50 relapse remitting MS patients and 50 healthy controls by TaqMan Real time PCR. All the patients were negative for HLA-DRB1*15 susceptible allele, normal serum vitamin D, responder to Interferon beta. All the health individuals were matched to patients. Also, we tried to find correlation between TRAIL gene expression and clinical characteristics of patients. RESULTS: No statistically significant difference was found in TRAIL mRNA expression between MS patients and controls (p> 0.05). There was no correlation in the TRAIL expression and age of onset, disease duration and Expanded Disability Status Scale of Kurtzke (EDSS). As IFN-b may have stimulatory effects on immunoregulatory function of TRAIL and all of our patients were treated with interferon beta and were responder, it lead to no significant change in TRAIL expression. We suggest comparing between responders and non-responders should be investigated.