Polymorphisms in CD14 Gene May Modify Soluble CD14 Levels and Represent Risk Factors for Multiple Sclerosis.

BACKGROUND: Besides the central role of the adaptive immune system, a disturbance of innate immune system is also suggested to be involved in the pathogenesis of multiple sclerosis (MS). CD14, a receptor upregulated in activated microglia, is known to be an essential mediator of inflammation in innate immune responses. Therefore, in this study we aimed to assess possible roles of CD14-159 and -260 gene polymorphisms in MS susceptibility and the effects of those polymorphisms to its protein producing capacity in Iranian population. METHODS: In this case control study, CD14-159 and -260 polymorphisms were genotyped using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) in 200 MS patients and 200 healthy controls matched in age and gender. Serum levels of soluble CD14 (sCD14) was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: There were significant differences in genotype distribution of CD14-159 and -260 polymorphisms between patients and controls (P = 0.01, for-both). Mean serum level of sCD14 was significantly higher in MS patients than in control subjects (3340.30 ± 612.50 ng/ml vs 2353.73 ± 539.07 ng/ml; P < 0.01). CONCLUSION: In summary, we conclude that CD14-159 and -260 polymorphisms are associated with the risk of MS in Iranian population and affects CD14 promoter activity, thereby regulating CD14 expression. Furthermore, our study provides preliminary evidence for the activation of innate immunity in the pathogenesis of MS. In addition, the findings of the present study suggest serum level of sCD14 as candidate biomarker of MS severity.

TNF-alpha Production by Peripheral Blood Monocytes in Multiple Sclerosis Patients and Healthy Controls.

BACKGROUND: Aberrant immune responses are evident in the pathogenesis of multiple sclerosis (MS) and it has been proposed that the spectrum of cytokines influence disease outcomes. Leptin and lipopolysaccharide (LPS) of Gram-negative bacteria are both potent cellular stimulators for production of pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α). The aim of this study was to compare the TNF-α production by peripheral blood monocytes from MS patients with healthy controls. METHODS: Peripheral blood samples were stimulated with LPS or leptin. After blocking the Golgi apparatus, intracellular cytokine production was assessed using a monoclonal antibody against human TNF-α by the flow cytometry technique. Moreover, plasma level measurement of cytokines was performed using enzyme-linked immunosorbent assay (ELISA). RESULTS: Intracellular levels of TNF-α were 16.80 ± 8.21 and 16.52 ± 8.23in MS patients and healthy controls which showed no statistically significant difference between them (p = 0.850). Leptin-stimulated and LPS-stimulated TNF-α production showed no significant difference between MS patients and the control group (p = 0.263 and p = 0.191, respectively). However, after treatment with leptin, a weak significant difference was shown between cases and control group (p = 0.049). There were significant differences between cases and controls regarding serum levels of IL-6 and Toll-like receptor-4 (TLR-4) before and after stimulation with leptin and LPS, separately (p < 0.05). CONCLUSION: Taken together, we cannot definitely conclude that TNF-α does not play an important role in pathogenesis of MS. However, other characteristics of monocyte activation such as IL-6 or TLRs can elucidate implication of peripheral blood monocytes in MS pathogenesis.

A Novel Approach to Discriminate Subgroups in Multiple Sclerosis.

Multiple sclerosis (MS) is an autoimmune disease of central nervous system. Since different types of immune cells are involved in MS pathogenesis, in this study we aimed to evaluate serum levels of several immunological components including soluble CD4 (sCD4), sCD8, sCD163, and immunoglobulins as markers of activity of T-cells, macrophages, and B-cells in different types of MS. Serum levels of sCD4, sCD8, and sCD163 of patients with relapsing-remitting MS (RRMS, n=61), primary progressive MS (PRMS, n=31), secondary progressive MS (SPMS, n=31), clinical isolated syndrome (CIS, n=31) and neuromyelitis optica (NMO, n=31), and healthy controls (n=49) were measured using enzyme-linked immunosorbent assay (ELISA). Serum levels of Ig-G, Ig-M, and Ig-A were determined using nephelometric technique. Serum levels of sCD4, sCD8, sCD163, Ig-G, Ig-M, and Ig-A were significantly different in five groups of cases (p<0.05). Furthermore, application of stepwise method of discriminant analysis yielded 4 significant discriminant functions of classification due to the presence of six levels of categorical variables in the analysis. The most important function explained 85.5% of the total variance with the correlation value of 0.79. Taken together, our preliminary analysis suggests that although we found some functions to discriminate most of the patients, further studies will be required to individuate immunological markers characterizing the different type of MS including RRMS, PPMS, SPMS, CIS and NMO as proved by the data on sCD4, sCD163, Ig-M, and Ig-G in blood.

Increased serum level of IL-37 in patients with multiple sclerosis and neuromyelitis optica.

Multiple sclerosis (MS) is a common autoimmune disease of central nervous system in which neurodegenerative and inflammatory mechanisms cause alternate neurological impairments. Many inflammatory and anti-inflammatory cytokines were suggested as contributor in MS pathogenesis, and the balance between these opposing cytokines can regulate MS severity. IL-37, an anti-inflammatory cytokine, is the most recently identified member of IL-1 family, which acts as a natural inhibitor of innate immunity. However, the role of IL-37 in MS has not investigated so far. Therefore, in this study, we aimed to measure serum level of IL-37 in patients with relapsing remitting multiple sclerosis (RRMS) and neuromyelitis optica (NMO). In a case-control study, plasma was collected from healthy controls (n = 49) and also patients with RRMS (n = 122) and NMO (n = 31). Serum level measurement of IL-37 was performed using enzyme-linked immunoassay (ELISA) method. The serum levels of IL-37 were 247.46 ± 74.02 and 312.00 ± 86.72 and 114.63 ± 20.58 in RRMS and NMO patients and healthy controls, respectively, showing statistically significant difference between them (P = 0.00). Furthermore, we found a positive correlation between the serum levels of IL-37 and EDSS of patients (r = +0.31 and P = 0.00). In summary, the serum level of IL-37 was found to be significantly increased in MS patients compared to healthy controls. Furthermore, the mean serum level of IL-37 was correlated with disease severity. This suggests that IL-37 may be part of a feed-back loop to control underlying inflammation in MS pathogenesis. However, further studies will be required to indicate exact role of IL-37 in the MS pathomechanisms.