This study aims to evaluate the effects of treatment with IFN-β 1α on the expressions of NLRP3, NLRP1, NLRC4, and AIM2, as inflammasomes, and caspase-1, IL-1β, and IL-18, as the downstream molecules of inflammasomes, in a population of Iranian multiple sclerosis (MS) patients. In this study, 30 MS patients (22 women and 8 men) participated. Before receiving any medication and 6 months after treatment with standard doses of IFN-β 1α 30 mcg injected intramuscularly once a week, blood samples were taken and then the leukocytes isolated, total RNAs extracted, and complementary DNAs (cDNAs) synthesized. Gene expressions of NLRP3, NLRP1, NLRC4, AIM2, and ASC were evaluated at messenger RNA (mRNA) levels using real-time PCR method; for assessing caspase-1 at protein level, the Western blot method was used. The amounts of IL-1β and IL-18 were measured in plasma using enzyme-linked immunosorbent assay method. Analysis of the results before and after therapy with IFN-β 1α in all patients shows significantly decreased expressions of NLRP3, NLRC4, and AIM2. The plasma levels of IL-1β, after treatment with IFN-β 1α, were significantly decreased in the MS patients. Based on our results, it appears that NLRP3, NLRC4, and AIM2 play critical roles in the progression of MS, probably by mediating Th1 and Th17 responses. It seems that decreased expression of IL-1β is related to decreased production and also functions of inflammasomes.

Non-viral gene delivery methods are considered due to safety and simplicity in human gene therapy. Since the use of cationic peptide and niosome represent a promising approach for gene delivery purposes we used recombinant fusion protein and cationic niosome as a gene carrier. A multi-domain fusion protein including nuclear localization motif (NLS) and two DNA-binding (Mu) domains, namely NLS-Mu-Mu (NMM) has been designed, cloned and expressed in E. coli DE3 strain. Afterward, the interested protein was purified by affinity chromatography. Binary vectors based on protein/DNA and ternary vectors based on protein/DNA/niosome were prepared. Protamine was used as a control. DNA condensing properties of NMM and protamine were evaluated by various experiments. Furthermore, we examined cytotoxicity, hemolysis and transfection potential of the binary and ternary complexes in HEK293T and MCF-7 cell lines. Protamine and Lipofectamine™2000 were used as positive controls, correspondingly. The recombinant NMM was expressed and purified successfully and DNA was condensed efficiently at charge ratios that were not harmful to cells. Peptidoplexes showed transfection efficiency (TE) but ternary complexes had higher TE. Additionally, NMM ternary complex was more efficient compared to protamine ternary vectors. Our results showed that niosomal ternary vector of NMM is a promising non-viral gene carrier to achieve an effective and safe carrier system for gene therapy.

Objectives: Interferon-β 1a (IFN-β 1a) is a common strategy therapy for multiple sclerosis (MS) with unknown mechanisms. S100A12 (S100 calcium-binding protein A12) is a damage-associated molecular pattern molecule which binds to its receptor, RAGE (receptor for advanced glycation end products), and activates nuclear factor-κB (NF-κB). NF-κB is transcribed from proinflammatory molecules, which may participate in the pathogenesis of MS. Therefore, the aims of this study were to compare mRNA levels of S100A12, RAGE, and NF-κB in newly diagnosed MS patients with healthy controls and determine whether IFN-β 1a therapy affects the expression of the molecules. Methods: S100A12, RAGE, and NF-κB mRNA levels in 30 new cases of untreated MS patients and 35 healthy controls were evaluated using the real-time PCR technique. The mRNA levels were also evaluated in the MS patients after 6 months of IFN-β 1a therapy. Results: S100A12, RAGE, and NF-κB mRNA levels were significantly decreased in the new cases of untreated MS patients in comparison to healthy controls. IFN-β 1a therapy results in upregulation of RAGE in MS patients, but not S100A12 and NF-κB. Conclusions: It appears that S100A12 participates in the pathogenesis of MS, and it seems that IFN-β 1a modulates immune responses in an S100A12-independent manner. Based on the reported anti-inflammatory effects of RAGE, it seems that RAGE may be considered as a mechanism by IFN-β 1a to modulate immune responses. NF-κB is produced permanently in the human cells and is inactive in the cytoplasm; therefore, the effects of IFN-β 1a may be related to its functions rather than expressions.

The aims of this study were to compare mRNA levels of melanoma differentiation‐associated protein 5 (MDA5) and retinoic acid‐inducible gene 1 (RIG‐1) in multiple sclerosis (MS) patients in comparison to the healthy controls as well as investigating the effects of IFN‐β 1a on the expression of these molecules. In this study, mRNA levels of MDA5 and RIG‐1 in peripheral leukocytes of 30 new cases of MS patients and 35 healthy controls were evaluated using the real‐time‐PCR method. mRNA levels of MDA5 and RIG‐1 were determined in the MS patients 6 months after treatment with standard doses of IFN‐β 1a. mRNA levels of MDA5 and RIG‐1 were significantly decreased in the MS patients in comparison to the healthy controls. The analysis also revealed that IFN‐β 1a therapy leads to the upregulation of RIG‐1, but not MDA5, in the total MS patients and the female group. MS patients suffer from insufficient expression of MDA5 and RIG‐1, and IFN‐β 1a therapy results in the upregulation of RIG‐1 in the patients, especially in the female patients. Thus, it seems that IFN‐β 1a not only decreased pathogenic inflammatory responses but also modulated the expression of RIG‐1 to protect the patients from infectious diseases and upregulation of IFN‐I in a positive feedback.

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