Therapeutic Efficacy of Mesenchymal Stem Cells and Mesenchymal Stem Cells-derived Neural Progenitors in Experimental Autoimmune Encephalomyelitis

BACKGROUND AND OBJECTIVES: The goal of treatment for MS is to reduce the inflammation and induce the regeneration of degenerated axons. Considering the anti-inflammatory and regenerative capacity of mesenchymal stem cell (MSCs), in this study the therapeutic efficacy of allogeneic MSCs and MSCs-derived neural progenitor cells (MSCs-NPs) was investigated in cellular therapy of chronic experimental autoimmune encephalomyelitis (EAE). METHODS AND RESULTS: MSCs, MSCs-NPs and MSCs+MSCs-NP were administered intravenously to EAE mice on days 22, 29, and 36 post immunization. The levels of cytokines and PGE2 in sera or supernatant of in vitro cultured splenocytes derived from treated mice were measured by ELISA. The results of this study showed that in comparison to MSCs monotherapy, MSCs-NPs administration had a more profound capability of inhibiting the proliferation of pathogenic MOG35–55-specific T cells, decreasing IFN-γ production and increasing anti-inflammatory IL-10 cytokine production. These findings could be explained by higher ability of in vitro cultured MSCs-NPs in production of PGE2 compared to MSCs. In line with these findings, while the administration of MSCs and MSCs-NPs significantly decreased the clinical scores of EAE in comparison with the untreated EAE group, MSCs-NPs were significantly more efficient in reducing clinical score compared to MSCs. Of interest, combined therapy with MSCs and MSCs-NPs did not provide any benefit over monotherapy with MSCs-NPs. CONCLUSIONS: In comparison to MSCs, allogenic MSCs-NPs are more potent in the attenuation of EAE.

Protective effect of edaravone against cyclosporine-induced chronic nephropathy through antioxidant and nitric oxide modulating pathways in rats

Background: Cyclosporine A [CsA] is an immunosuppressant with therapeutic indications in various immunological diseases; however, its use is associated with chronic nephropathy. Oxidative stress has a crucial role in CsA-induced nephrotoxicity. The present study evaluates the protective effect of edaravone on CsA-induced chronic nephropathy and investigates its antioxidant and nitric oxide modulating property

Methods: Male Sprague-Dawley rats [n=66] were distributed into nine groups, including a control [group 1] [n=7]. Eight groups received CsA [15 mg/kg] for 28 days while being treated. The groups were categorized as: - Group 2: Vehicle [n=10]. - Groups 3, 4, and 5: Edaravone [1, 5, and 10 mg/kg] [n=7 each]. - Group 6: Diphenyliodonium chloride, a specific endothelial nitric oxide synthase [eNOS] inhibitor [n=7]. - Group 7: Aminoguanidine, a specific inducible nitric oxide synthase [iNOS] inhibitor [n=7]. - Group 8: Edaravone [10 mg/kg] plus diphenyliodonium chloride [n=7]. - Group 9: Edaravone [10 mg/kg] plus aminoguanidine [n=7]. Blood urea nitrogen and serum creatinine levels, malondialdehyde, superoxide dismutase, and glutathione reductase enzyme activities were measured using standard kits. Renal histopathological evaluations and measurements of eNOS and iNOS gene expressions by RT-PCR were also performed. Data were analyzed using one-way analysis of variance [ANOVA] followed by Tukey's test [SPSS software version 18.0]

Results: Edaravone [10 mg/kg] significantly attenuated CsA-induced oxidative stress, renal dysfunction, and kidney tissue injury. Aminoguanidine improved the renoprotective effect of edaravone. Edaravone reduced the elevated mRNA level of iNOS, but could not alter the level of eNOS mRNA significantly

Conclusion: Edaravone protects against CsA-induced chronic nephropathy using antioxidant property and probably through inhibiting iNOS gene expression

Serum levels of monocytechemoattractant protein-1 correlate withpoor clinical grades in cerebralaneurysms

Background: Ruptured cerebral aneurysms [ICAs] are the most common non-traumaticcause of subarachnoid hemorrhage [SAH] that is associated with life threateningcomplications such as Vasospasm, Infarction, and Hydrocephalus [HCP]. The activeparticipation of macrophage/monocyte-mediated inflammatory response in thepathogenesis of cerebral aneurysm as labeled with Monocyte ChemoattractantProtein-1[MCP-1] is suggested

Objective: To measure the serum level of MCP-1 in rupturedCAs in different time intervals

Methods: We measured the serum levels of MCP-1 inSAH patients who had CAs and compared it with that of MCP-1 in two control groups:including patients with SAH without CAs, and the normal population of blood donors.We also measured the MCP-1 levels in patients with CAs one week afterward toevaluate the effect of treatment. Serum level of MCP-1 was measured by a commercial ELISA assay

Results: Mean serum MCP-1 level in patients with SAH and CAs was188.2168 Pg/ml and 331.3982 Pg/ml in the normal population. There was nostatistically significant difference between serum levels of MCP-1 on the first[mean=188.2168 Pg/ml] and 7th days after SAH onset [mean=171.8450 Pg/ml][p=0.739]. Serum level of MCP-1 increased significantly as Glasgow Coma Scaledecreased [p=0.078] and Hunt and Hess score increased [p=0.089]

Conclusion: Ourresults did not show an increasing MCP-1 serum level in patients with aneurysmalSAH. There was a relationship between poor clinical grade and MCP-1 levels inpatients with CAs. MCP-1 may be a local inflammatory marker for cerebral aneurysmswithout systemic manifestation

Association between tumor necrosis factor- alpha-308 G/A polymorphism and multiple sclerosis: a systematic review and meta-analysis

Multiple sclerosis [MS] is a complex polygenic disease in which gene-environment interactions are important. A number of studies have investigated the association between tumor necrosis factor-alpha [TNF-alpha] -308 G/A polymorphism [substitution G[rightwards arrow]A, designated as TNF1 and TNF2] and MS susceptibility in different populations, but the results of individual studies have been inconsistent. Therefore, performing a systematic review and meta-analysis of the published studies is desirable. We sought to quantitatively summarize the association between TNF-alpha-308 G/A polymorphism and MS. The Medline and Scopus databases were searched to identify potentially relevant case-control studies published in English journals up to January 2010. A meta-analysis of these studies was performed. Summary odds ratios [ORs] and 95% confidence intervals [CIs] were calculated under fixed and random effects models. Twenty-one eligible studies, comprising 2880 patients with MS and 3579 controls, were included in the meta-analysis. The overall pooled ORs [95%CI] for TNF2 versus TNF1 and TNF2 carriers [2/2+2/1] versus non-carriers [1/1] were 1.02 [0.86-1.21] and 0.99 [0.8-1.24], respectively. In the European populations, the pooled ORs [95%CI] for TNF 2/1 versus 1/1 were 0.85 [0.73-0.98], which was statistically significant. However, the other results did not support this finding. The pooled ORs [95%CI] for TNF 2/1 versus 1/1 and TNF 2/2 versus 2/1 were not statistically significant in the overall population. In addition, the pooled ORs for TNF2/2 versus TNF2/1+1/1 and TNF2/2 versus TNF1/1 were not statistically significant. Our meta-analysis does not support the role of TNF-alpha -308 G/A polymorphism in developing MS

Evaluation of the immunomodulatory effect of curdlan on maturation and function of mouse spleen-derived dendritic cells

T helper 1 and T helper 17 cells play important roles in immunity against foreign invaders. Differentiation of these Th subsets is affected by state of maturation and cytokines that are produced by dendritic cells [DCs]. Curdlan is a linear [1-3]-beta- glucan and has shown activity against tumors and infectious agents. This study aims to investigate whether curdlan plays its role through affecting the maturation and cytokine production by DCs. DCs were isolated from the spleen of BALB/c mice by MACS method. After an overnight culture of DCs in the presence of curdlan, the expression levels of CD40, CD86, and MHC-II molecules were determined by flow cytometry. The production of cytokines involved in Th1 and Th17 cell differentiation [IL-12 and IL-6, respectively] was also evaluated by ELISA. Lipopolysaccharide [LPS] treated and untreated cells were considered as positive and negative controls, respectively. The results of this study did not show a significant difference in the levels of surface expression of CD40 [p=0.82], CD86 [p=0.79], and MHC class II [p=0.84] molecules upon exposure to curdlan. However, LPS increased the intensity of CD40 expression on dendritic cells [p=0.04]. In addition, it was revealed that curdlan-exposed DCs are not able to produce a significant amount of IL-6 and IL-12 cytokines. Conversely, LPS-treated DCs were able to make a significant amount of IL-12 [p=0.005]. The results of the present study suggest that curdlan has no effect on Th1 or Th17 differentiation while LPS may induce Th1 deviation by induction of CD40 expression and IL-12 production.

Anti-cardiolipin and anti-neutrophil cytoplasmic antibodies in Iranian patients with behcet's disease

The prevalence of anti-Neutrophil Cytoplasmic Antibodies [ANCAs] and anti-Cardiolipin Antibodies [anti-CL Ab] in Behcet's Disease [BD] and also their roles in vascular involvement is controversial. To assess the prevalence of ANCAs and anti-CL Ab as well as their correlations with clinical manifestations in Iranian patients with BD. In this case/control study, the sera from 88 patients with BD and 88 healthy controls were evaluated. The levels of ANCAs and anti-CL Ab were measured using indirect ELISA method. The levels of anti-CL, anti-PR3 and anti-MPO [Myeloperoxidase] IgG autoantibodies between BD patients and healthy controls were not statistically different [p=0.21, p=0.28 and p=0.74, respectively]. In addition, there were no significant deferences between BD patients with and without vascular involvement in the levels of anti-CL [1.42 +/- 1.24 GPLU/ml and 1.58 +/- 1.18 GPLU/ml, respectively; p=0.71], anti-PR3 [0.0 +/- 0.0 U/ml and 0.08 +/- 0.27 U/ml, respectively; p=0.10] and anti MPO [0.48 +/- 0.23 U/ml and 0.52 +/- 0.22 U/ml, respectively; p=0.41] IgG autoantibodies. Nevertheless, mean titer of anti-CL IgG was higher in male patients with skin rash than those without skin rash [2.2 +/- 0.88 GPLU/ml and 1.11 +/- 1.22 GPLU/ml, respectively; p=0.017]. While anti-CL, antiPR3 and anti-MPO IgG autoantibodies do not play a major role in susceptibility to BD or pathogenesis of vascular involvement in our patients, anti-CL Ab might be involved in skin lesion development in Iranian male BD patients. However, the results should be confirmed in other studies

Interleukin-8-251 A/T and CXCR2 +1208 C/T genes polymorphisms in chronic rhinosinusitis

IL-8 is one of the pro-inflammatory cytokines which can play an essential role in the pathogenesis of chronic rhinosinusitis [CRS] as well as nasal polyposis [NP]. The ability of individuals in producing IL-8 is partially determined by IL-8-251 A/T polymorphism. Hence, the aim of the present study was to investigate the association between IL-8-251 A/T and CXCR2 +1208 C/T genes polymorphisms and susceptibility to CRS and NP. Two hundred and forty five CRS patients and 204 healthy controls were included in this study. CRS patients were categorized by the existence or absence of NP. IL-8 promoter-251 A/T and CXCR2 +1208 C/T gene polymorphisms were genotyped via the allele specific PCR [AS-PCR] method. While no remarkable difference was demonstrated between patients and controls for both CXCR2 +1208 C/T and IL-8 -251 A/T polymorphisms, a significant increase in IL-8-251 AA genotype was detected in CRS patients with NP compared to those without it [29.3% and 16.2%, respectively; P=0.03]. Interestingly, this association got far stronger when only non-asthmatic CRS patients were taken into consideration [P=0.001]. The results of the present study indicate that the inheritance of IL-8-251 Aallele is associated significantly with NP development in CRS patients. Therefore, NP formation might be a result of the exposure to an intense inflammatory environment, which is more likely in genetically susceptible CRS patients

Effect of Fas-670 A/G gene polymorphism on corneal allograft endothelial rejection

Human cornea expresses functional Fas-ligand capable of killing Fas+ activated lymphocytes. Fas expression is partly regulated by -670 A/G polymorphism in the promoter region of Fas gene. The aim of the present study is to determine the association between Fas-670A/G polymorphism and survival of corneal transplantation. In 276 graft recipients who mainly underwent penetrating keratoplasty because of keratoconus, bullous keratopathy and corneal opacity, Fas -670 A/G polymorphism was determined by allele specific oligonucleotide polymerase chain reaction [ASO-PCR] techniques. There was no statistically significant relationship between Fas -670 A/G polymorphism and rejection episode [p=0.35]. Moreover, the relationship between this polymorphism and rejection episode outcome [transplant recovery vs failure] was not statistically significant [p=0.13]. The results of the present study show no significant correlation between corneal graft rejection, rejection recovery and Fas -670A/G gene polymorphism

Cloning and expression of Leishmania major superoxide dismutase B1: a potential target antigen for serodiagnosis of Leishmaniasis

Leishmaniasis- a neglected public health problem- is a group of diseases affecting an estimated 12 million people worldwide. In the present study, recombinant Leishmania major superoxide dismutase B1 [rLmSODB1] has been utilized as a potential antigen for the serodiagnosis of human cutaneous [CL] and visceral leishmaniasis [VL] in the endemic regions of southern part of Iran. Additionally, the sensitivity and specificity of ELISA-based serodiagnosis using rLmSODB1 and the soluble Leishmania antigen [SLA] were compared. For the first time, rLmSODB1 has been cloned successfully and used for ELISA-based serodiagnosis. Sera from 30 CL and 24 VL cases were included in this study. Additional studies were also done for the evaluation of cross-reactivity using sera from 41 endemic controls including normal endemic donors [n= 20], systemic lupus erythematosus patients [n=5], rheumatoid arthritis patients [n= 5], and patients with tuberculosis [n=11]. Analysis indicated that rLmSODB1 was recognized by62.5% and 13.3% of sera from patients with VL and CL, showing a sensitivity of 72.7% and 53.6%, respectively. However 95.8% of VL and 30% of CL sera reacted with SLA, revealing sensitivities of 96% and 58.8%, respectively. Additionally, from 41 sera collected either from healthy subjects or patients affected with other diseases, 97.5% were negative with SLA or rLmSODB1 [specificity 97.6%]. These results show that rLmSODB1 almost does not react with sera from patients with tuberculosis and autoimmune diseases and may be considered as a candidate antigen for the specific immunodiagnosis of visceral leishmaniasis

NOS2A gene polymorphism and its association with nitric oxide production in peripheral blood leukocytes of normal individuals

In peripheral blood polymorphnuclear and mononuclear cells Nitric Oxide [NO] could be synthesized by an enzyme called inducible NO synthase [iNOS]. iNOS gene [NOS2A] is located on chromosome 17 at position 17q11.2-q12. NO is released during inflammatory responses. In the present studies the frequency of NOS2A gene polymorphisms and their effects on NO production were investigated in the Peripheral Blood polymorphonuclear and mononuclear cells of normal individuals. In this study the frequency of NOS2A gene polymorphisms at positions -1659 C/T and +150 C/T of 232 normal subjects were investigated using PCR-Allele specific and PCR-RFLP methods, respectively. To study the effect of 1659C/T and +150C/T polymorphisms on NO production, polymorphonuclear and mononuclear cells of peripheral blood of 92 normal subjects were isolated and then stimulated by E.coli culture supernatants [ATCC 25922] for induction of iNOS enzyme and NO production. After 24h, the level of NO production in the culture supernatant were measured by Griess reaction. Polymorphisms as mentioned above were also studied in these normal cases. The results showed no significant difference in the level of NO production of various genotypes in the polymorphonuclear and mononuclear cells of peripheral blood of normal subjects. Our results indicated no significant correlation between NOS2A genotypes and NO production. No significant difference was observed between Gambia and China normal population and normal subjects of this study in NOS2A- 1659 C/T and +150 C/T polymorphisms. In Iran these differences are due to the genetic and ethnic differences among the studied populations, which indicates the importance of NOS2A polymorphism in the NO production, suggesting further studies in other ethnic groups

Aqueous humor levels of soluble fas and fas-ligand in patients with primary open angle and pseudoexfoliation glaucoma

Glaucoma is one of the most common causes of blindness and is usually associated with elevated intraocular pressure. In patients with primary open angle glaucoma the number of trabecular meshwork cells is decreased. Death of the trabecular meshwork cells may be a result of apoptosis. To investigate the aqueous humor levels of soluble Fas [sFas] and Fas-Ligand [sFasL] in glaucomatous patients. Concentration of sFas and sFasL were measured by ELISA in 41 eyes with glaucoma [21 with pseudoexfoliation and 20 with primary open angle glaucoma] and 39 eyes with cataract as controls. The sFas concentration was lower in the primary open angle than the pseudoexfoliation glaucoma and the cataract groups [p=0.002 and p= 0.004, respectively]. The sFasL level did not show any significant difference in the three groups. A lower level of sFas may provide proper microenvironment for increased apoptosis of trabecular meshwork cells in primary open angle glaucoma

Investigation of Fc gamma RIIA and Fc gamma RIIIA polymorphism in multiple sclerosis: a case control study

Multiple Sclerosis [MS], the most common demyelinating disease of the CNS, is immunologically mediated in genetically susceptible individuals. Receptors for the Fc fragment of IgG [Fc gamma R] might induce inflammatory responses through linking the humoral and cellular immune responses by targeting immune complexes to effector cells. Polymorphisms in some Fc gamma R genes are associated with various infectious and autoimmune diseases, probably due to their effects on different binding capacities of encoded receptors for IgG containing immune complexes. To investigate the importance of Fc gamma R polymorphisms in susceptibility to MS. One hundred and fifty MS patients and 136 age and sex matched controls were genotyped for Fc gamma RIIA and Fc gamma RIIIA gene polymorphisms using PCR-RFLP method. The allelic and genotypic frequencies of the Fc gamma RIIA and Fc gamma RIIIA did not differ significantly between the MS patients and controls. There was no association between allelic polymorphism of Fc gamma RIIIA and severity of disease based on Expanded Disability Status Scale [EDSS] score. However, significant association between inherited Fc gamma RIIA genotype and disease activity [p=0.001] or progression index was revealed [p=0.014]. EDSS values showed that Fc gamma RIIA [H/H] and [H/R] genotypes were associated with a lower EDSS score in relapsing-remitting MS and in the total MS population [P=0.001] but not [R/R] genotype. Considering the detrimental role of autoantibodies in the pathogenesis of MS, our results suggest that the inherited Fc gamma RIIA alleles could affect the severity of MS by influencing the clearance rate of immune complexes and autoantibodies. The results of the present study add the Fc gamma RIIA gene to the gene networks which determine the severity of MS in southern Iran

Evaluation of cellular immune response against purified antigen 85 in patients with tuberculosis

Tuberculosis [TB] remains an important health problem throughout the world. Despite its significance in public health, mechanisms of protective immunity against Mycobacyerium Tuberculosis in humans have not yet been understood. To evaluate cell mediated immune response against purified Ag 85, PPD and Phytohemagglutinin [PHA] in patients with tuberculosis and healthy tuberculin positive and negative individuals. Thirty patients with tuberculosis and 60 healthy tuberculin skin test positive and negative volunteers were participated in this study. Cell mediated immunity was assessed by measuring [[3]H]-thymidine uptake and detection of IFN-gamma in the culture supernatant using commercial ELISA test. In the present study, we showed that IFN-gamma production and cell proliferation response to Ag 85 were significantly higher in tuberculin positive than tuberculin negative individuals [P<0.01]. Among tuberculous patients, IFN-gamma production and cell proliferative responses to Ag 85 was significantly lower in contrast to healthy tuberculin positive individuals [P<0.01]. In addition, IFN- gamma response in patients with cavitary tuberculosis was lower than patients without cavitation [P<0.05]. Based on the higher cell mediated immune responses to Ag 85 in healthy tuberculin positive volunteers compared to patients [especially with advanced disease], purified Ag 85 can be used as a sensitive marker for analysis of immune responses in tuberculosis

T-cell tolerance following bacterial glutamic acid decarboxylase [GAD] feeding in streptozotocin-induced diabetes

Autoimmune type 1 diabetes mellitus is caused by T-cell mediated immune destruction of the insulin-producing a-cell in pancreatic islets of Langerhans. Specificity of the auto-antibodies and of the auto-reactive T-cells has been investigated, in which several auto-antigens were proposed. To determine whether glutamic acid decarboxylase [GAD] feeding would induce oral tolerance of either T-cell or B-cell compartment in streptozotocin [STZ] diabetic rats. Rats in the experimental group were fed 2 mg/kg of GAD [extracted from Escherichia coli] 14 days before intra-peritoneal injections of streptozotocin [30 mg/kg body weight for 5 consecutive days]. Two control groups were considered: diabetic control group, which underwent STZ injections without receiving GAD, and normal control group. Systemic response was compared between the three groups. T-cells response was assessed by a proliferation assay of spleen cells and those of the B-cells by enzyme-linked immunosorbent assay [ELISA] for anti-GAD specific antibodies in serum. Compared with the diabetic control group, a significant reduction was observed only in the proliferative response of spleen cells, but not in the level of anti-GAD antibody. GAD feeding induces systemic T-cell tolerance in STZ-induced diabetes

Interleukin-10 promoter polymorphisms and breast cancer risk in Iranian women

IL-10 is an anti-inflammatory cytokine which is involved in tumorigenesis. Over production of IL-10 and elevated number of IL-10 generating mononuclear cells in breast tumor tissue has already been shown. To determine the association of IL-10 promoter polymorphisms with increased risk of breast cancer and its association with breast cancer prognostic factors. Peripheral blood samples from 275 female breast cancer patients and 320 cancer free controls were used to detect three single nucleotide polymorphisms in IL-10 promoter region [-1082, -819, -592] by PCR method. The frequency of genotypes and alleles of three mentioned regions of IL-10 promoter and their haplotypes [GCC, ATA, and ACC] showed no statistically significant difference between patients and controls. In the case of prognostic factors, progesterone receptor [PR] status exhibited significant relation with -1082 genotypes [P=0.03] and haplotypes [P=0.02]. -1082 AA genotype was associated with negative PR expression whereas AG and GG genotypes of this site were positively associated with PR expression. Similarly GCC haplotype correlated with positive PR expression and ATA and ACC with negative PR expression. The data of this study showed that IL-10 promoter gene polymorphisms may not be considered as one of the risk factors for breast cancer in Iranian patients