Dual effects of cell free supernatants from lactobacillus acidophilus and lactobacillus rhamnosus GG in regulation of MMP-9 by up-regulating TIMP-1 and down-regulating CD147 in PMA- differentiated THP-1 cells

Objective: recent studies have reported dysregulated expression of matrix metalloproteinases [MMPs], especially MMP-2, MMP-9, tissue inhibitor of metalloproteinase-1, -2 [TIMP-1, TIMP-2], and extracellular matrix metalloproteinase inducer [EMMPRIN/CD147] in activated macrophages of patients with inflammatory diseases. Therefore, MMP-2, MMP-9, and their regulators may represent a new target for treatment of inflammatory diseases. Probiotics, which are comprised of lactic acid bacteria, have the potential to modulate inflammatory responses. In this experimental study, we investigated the anti-inflammatory effects of cell-free supernatants [CFS] from Lactobacillus acidophilus [L.acidophilus] and L. rhamnosus GG [LGG] in phorbol myristate acetate [PMA]-differentiated THP-1 cells

Materials and Methods: in this experimental study, PMA-differentiated THP-1 cells were treated with CFS from L. acidophilus, LGG and uninoculated bacterial growth media [as a control]. The expression of MMP-2, MMP-9, TIMP-1, and TIMP-2 mRNAs were determined using real-time quantitative reverse transcription polymerase chain reaction [RTPCR]. The levels of cellular surface expression of CD147 were assessed by flow cytometry, and the gelatinolytic activity of MMP-2 and MMP-9 were determined by zymography

Results: our results showed that CFS from both L. acidophilus and LGG significantly inhibited the gene expression of MMP-9 [P=0.0011 and P=0.0005, respectively], increased the expression of TIMP-1 [P<0.0001], decreased the cell surface expression of CD147 [P=0.0307 and P=0.0054, respectively], and inhibited the gelatinolytic activity of MMP-9 [P=0.0003 and P<0.0001, respectively] in PMA-differentiated THP-1 cells. Although, MMP-2 expression and activity and TIMP-2 expression remained unchanged

Conclusion: our results indicate that CFS from L. acidophilus and LGG possess anti-inflammatory properties and can modulate the inflammatory response

role of M2000 as an anti-inflammatory agent in toll-like receptor 2/microRNA-155 pathway

Background: M2000 is a newly designed and safe Non-Steroidal Anti-Inflammatory Drug [NSAID]. The aim of this study was to assess the effects of M2000 on expression levels of Suppressor of Cytokine Signaling-1 [SOCS-1] and Src Homology-2 domain containing inositol-5'-phosphatase 1 [SHIP1] proteins via Toll-Like Receptor [TLR] 2/microRNA-155 pathway

Methods: HEK293 TLR2 cell line and Peripheral Blood Mononuclear Cells [PBMCs] were treated by different concentrations of M2000 in MTT assay. RNA was extracted by miRN easy Mini kit. Then, cDNA was synthesized and the expression levels of SOCS1, SHIP1 and miRNA155 were evaluated by Quantitative Real time PCR

Results: Our results showed that M2000 significantly increased the expression levels of SOCS1 and SHIP-1 in Lipopolysachride [LPS]-treated and non-treated cells. Moreover, M2000 decreased expression level of miR-155 in LPS treated PBMCs

Conclusion: M2000 can be used as NSAID in LPS induced inflammation and decrease inflammatory cytokines production by targeting SOCS1, SHIP1 and miR-155 in autoimmune and inflammatory diseases

The Effect of Lactobacillus acidophilus PTCC 1643 on Cultured Intestinal Epithelial Cells Infected with Salmonella enterica serovar Enteritidis

OBJECTIVES: Gastrointestinal disorders caused by Salmonella enterica serovar Enteritidis (SesE) are a significant health problem around the globe. Probiotic bacteria have been shown to have positive effects on the immune responses. Lactobacillus acidophilus was examined for its capability to influence the innate immune response of HT29 intestinal epithelial cells towards SesE. The purpose of this work was to assess the effect of L. acidophilus PTCC 1643 on cultured intestinal epithelial cells infected with SesE. METHODS: HT29 cells were cultured in Roswell Park Memorial Institute medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. The cells were treated with L. acidophilus PTCC 1643 after or before challenge with SesE. At 2 and 4 hours post-infection, we measured changes in the expression levels of TLR2 and TLR4 via real-time polymerase chain reaction. RESULTS: Treatment with L. acidophilus inhibited SesE-induced increases in TLR2 and TLR4 expression in the infected HT29 cells. Moreover, the expression of TLR2 and TLR4 in cells that were pretreated with L. acidophilus and then infected with SesE was significantly higher than that in cells infected with SesE without pretreatment. Taken together, the results indicated that L. acidophilus had an anti-inflammatory effect and modulated the innate immune response to SesE by influencing TLR2 and TLR4 expression. CONCLUSION: Our findings suggested that L. acidophilus PTCC 1643 was able to suppress inflammation caused by SesE infection in HT29 cells and reduce TLR2 and TLR4 expression. Additional in vivo and in vitro studies are required to further elucidate the mechanisms underlying this anti-inflammatory effect.

Dysbiosis of fecal microbiota and high frequency of citrobacter, klebsiella spp., and actinomycetes in patients with irritable bowel syndrome and gastroenteritis

Aim: This study was aimed to characterize putative differences of fecal microbiota between irritable bowel syndrome[IBS] and gastroenteritis patients and healthy controls

Background: New evidence proposed that gut microbiota has a deep effect on the balance between health and disease

Patients and methods: The presence of Clostridium difficile, Campylobacter spp., Enterobacteriacea and Staphylococci weredetected in the samples using selective and specific culture media. Microscopic examination of the samples was done to detectActinomycetes, yeasts, Bifidobacteria, Fusobacterium spp., as well as white blood cells, red blood cells, mucus and epithelial cells

Results: Results of this study showed relatively higher frequency of Citrobacter spp., Lactobacilli, and Actinomycetes in theIBS patients. Elevated levels of WBC, RBC secretion, and increased amounts of Klebsiella, Escherichia coli and Citrobacterspp. were characterized in the patients with gastroenteritis compared with the control group

Conclusion: Depletion of gram positive cocci and gram negative bacilli also suggested dysbiosis of intestinal microbiotain these patients

role of inflammatory mediators in the pathogenesis of Alzheimer's disease

Alzheimer's disease [AD], a neurodegenerative disorder associated with advanced age, is the most common cause of dementia globally. AD is characterised by cognitive dysfunction, deposition of amyloid plaques, neurofibrillary tangles and neuro-inflammation. Inflammation of the brain is a key pathological hallmark of AD. Thus, clinical and immunopathological evidence of AD could be potentially supported by inflammatory mediators, including cytokines, chemokines, the complement system, acute phase proteins and oxidative mediators. In particular, oxidative mediators may actively contribute to the progression of AD and on-going inflammation in the brain. This review provides an overview of the functions and activities of inflammatory mediators in AD. An improved understanding of in?ammatory processes and their role in AD is needed to improve therapeutic research aims in the field of AD and similar diseases

live vector expressing HPV16 L1 generates an adjuvant-induced antibody response in-vivo

Background: The association between human papillomavirus [HPV] infections and cervical cancer has suggested the design of prophylactic and therapeutic vaccines against genital warts. The HPV capsid has made of two L1 and L2 coat proteins that have produced late in viral infections. Regarding to the recent studies, two commercial prophylactic vaccines have based on L1 viral like particles [VLPs] could strongly induce antibody responses, and protect human body from HPV infections. However, the use of these HPV vaccines has hindered due to their high cost and some limitations. Currently, among various vaccination strategies, live vector-based vaccines have attracted a great attention

Objectives: Herein, a non-pathogenic strain of the protozoan organism known as Leishmania tarentolae has utilized to induce potent humoral immunity in mice model

Materials and Methods: At first, cloning of HPV16 L1 gene into Leishmania expression vector has performed and confirmed by PCR and digestion with restriction enzymes. The promastigotes of Leishmania tarentolae [Ltar] have transfected with linearized DNA construct by electroporation. Protein expression has analyzed by SDS-PAGE and western blotting. Then, the immunogenicity of leishmania expressing L1 protein [Ltar-L1] has assessed in mice model

Results: Our data has indicated that subcutaneous immunization of mice with the recombinant L.tar-L1 has led to enhance the levels of IgG1 and lgG2a in comparison with control groups. Furthermore, there was no significant increase in antibody levels between two and three times of immunizations

Conclusions: The recombinant live vector was able to induce humoral immunity in mice without need of any adjuvant. However, further studies have required to increase its efficiency

significance of matrix metalloproteinases in the immunopathogenesis and treatment of multiple sclerosis

Multiple sclerosis [MS] is an autoimmune disease of the central nervous system [CNS]. The major pathological outcomes of the disease are the loss of blood-brain barrier [BBB] integrity and the development of reactive astrogliosis and MS plaque. For the disease to occur, the non-resident cells must enter into the immune-privileged CNS through a breach in the relatively impermeable BBB. It has been demonstrated that matrix metalloproteinases [MMPs] play an important role in the immunopathogenesis of MS, in part through the disruption of the BBB and the recruitment of inflammatory cells into the CNS. Moreover, MMPs can also enhance the cleavage of myelin basic protein [MBP] and the demyelination process. Regarding the growing data on the roles of MMPs and their tissue inhibitors [TIMPs] in the pathogenesis of MS, this review discusses the role of different types of MMPs, including MMP-2, -3, -7, -9, -12 and -25, in the immunopathogenesis and treatment of MS

Calcium intervention ameliorates experimental model of multiple sclerosis

Multiple sclerosis [MS] is the most common inflammatory disease of the CNS. Experimental autoimmune encephalomyelitis [EAE] is a widely used model for MS. In the present research, our aim was to test the therapeutic efficacy of Calcium [Ca] in an experimental model of MS. In this study the experiment was done on C57BL/6 mice. EAE was induced using 200 micro g of the MOG[35-55] peptide emulsified in CFA and injected subcutaneously on day 0 over two flank areas. In addition, 250 ng of pertussis toxin was injected on days 0 and 2. In the treatment group, 30 mg/kg Ca was administered intraperitoneally four times at regular 48 hour intervals. The mice were sacrificed 21 days after EAE induction and blood samples were taken from their hearts. The brains of mice were removed for histological analysis and their isolated splenocytes were cultured. Our results showed that treatment with Ca caused a significant reduction in the severity of the EAE. Histological analysis indicated that there was no plaque in brain sections of Ca treated group of mice whereas 4 +/- 1 plaques were detected in brain sections of controls. The density of mononuclear infiltration in the CNS of Ca treated mice was lower than in controls. The serum level of Nitric Oxide in the treatment group was lower than in the control group but was not significant. Moreover, the levels of IFN- gamma in cell culture supernatant of splenocytes in treated mice were significantly lower than in the control group. The data indicates that Ca intervention can effectively attenuate EAE progression

Immune reactivity of Brucella melitensis-vaccinated rabbit serum with recombinant Omp31 and DnaK proteins

Brucella melitensis infection is still a major health problem for human and cattle in developing countries and the Middle East. In this study, in order to screen immunogenic candidate antigens for the development of a Brucella subunit vaccine, a cytoplasmic protein [DnaK] and an outer membrane protein [Omp31] of B. melitensis were cloned, expressed in E.coli BL21 and then purified using Ni-NTA agarose. Immunized serum was prepared from a rabbit inoculated with attenuated B. melitensis. It was proved that immunized serum contains antibodies against recombinant Omp31 [rOmp31] and DnaK [rDnaK] by Western blot and ELISA assays. The results may suggest the importance of these proteins as subunit vaccines against B. melitensis as well as targets for immunotherapy

role of reactive oxygen species in immunopathogenesis of rheumatoid arthritis

Rheumatoid arthritis is a disease associated with painful joints that affects approximately 1% of the population worldwide, and for which no effective cure is available. It is characterized by chronic joint inflammation and variable degrees of bone and cartilage erosion. Oxygen metabolism has an important role in the pathogenesis of rheumatoid arthritis. Reactive oxygen species [ROS] are produced in many normal and abnormal processes in humans, including atheroma, asthma, joint diseases, aging, and cancer. TNF-? overproduction is thought to be the main contributor to increased ROS release in patients with RA. Increased ROS production leads to tissue damage associated with inflammation. The prevailing hypothesis that ROS promote inflammation was recently challenged when polymorphisms in Neutrophil cytosolic factor 1[Ncf1], that decrease oxidative burst, were shown to increase disease severity in mouse and rat arthritis models. It has been shown that oxygen radicals might also be important in controlling disease severity and reducing joint inflammation and connective tissue damage. In this review article, our aim is to clarify the role of ROS in immunopathogenesis of Rheumatoid arthritis

Suppression of telomerase activity by pyrimethamine: implication to cancer

Although pyrimethamine [Tindurin[TM]] appears to be effective in the prevention and treatment of some infectious diseases, very little information exists on its unpredictable properties. We design this study to evaluate its anti-tumoral effect on a model of cell line. The cytotoxic influence of Pyrimethamine on prostate cell line was investigated using an in vitro colometric assay. The potential modulatory effects on metastasis, apoptosis, and immortality characteristics of cells were assessed with gelatin zymography, terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling [TUNEL] assay and telomeric repeat amplification protocol, respectively. Cytotoxicity analysis of pyrimethamine revealed a dosedependent fashion. An apoptotic influence of pyrimethamine was also confirmed by data obtained from TUNEL assay. Dose-dependent inhibitory effect on matrix metalloproteinases [MMP] was seen in pyrimethamine. A potent inhibitory effect of pyrimethamine was also established by data achieved from TRAPeze telomerase detection kit. Collectively, as induction of apoptosis together with MMP and telomerase inhibition could be indicative of cancer treatment, pyrimethamine might be considered as a chemopreventative agent in cancer

In vitro assessment of bee venom effects on matrix metalloproteinase activity and interferon production

Controversial immunomodulatory properties of bee venom [BV] have provided an appropriate field for more investigation. The aim of present research was to verify the effects of honeybee venom on matrix metalloproteinase activity and interferon production as well as cell proliferation in monocyte and fibroblast cell lines. The monocyte and fibroblast cell lines [K562, HT-1080, WEHI-164] were used in order to assess proliferative response, interferon-1 production and matrix metalloproteinase-2 [MMP-2] activity. Australian BV [ABV] and Iranian BV [IBV] preparations at concentrations of 0.025, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, and 1microg/ml were added to each overnight cultured cells. In time course study, cells were treated each with ABV and IBV. In all cases supernatants were collected 24 hours after treatment. A sample of the each medium was used for zymography and interferons assay. Nontreated cells were used as controls. The production of IFN-alpha and IFN-beta in supernatant of cell cultures was assessed using enzyme linked immunoassay procedure. MMP-2 activity, as an inflammatory index, was evaluated using zymoanalysis method. The results of this study showed that, there were no significant difference between two sources of honey bee venoms when they were added to an identical cell line, whereas, the responses of various cell lines against bee venom were different. The increasing amounts of bee venom to human monocyte cell line [K562] revealed a significant increase in proliferative response. Our findings showed that the bee venom had no influence on IFN-alpha production in cell culture media, whereas, adding the BV to K562 cell line could significantly increase the production level of IFN-beta only on day 8 post-treatment. In addition the effect of bee venom on MMP-2 activity in both cell culture media, WEHI-164 and K562 was similar. The stimulatory effect of bee venom on MMP-2 activity occurred at low doses. In contrast, its inhibitory effect was seen at high concentrations. It is concluded that, honeybee venom affects on MMP-2 activity and interferon beta production as well as cell proliferation in a time and dose-dependent manner