IL-17 and IL-22 elicited by a DNA vaccine encoding ROP13 associated with protection against Toxoplasma gondii in BALB/c mice.

Toxoplasma gondii, an intracellular parasitic protozoan, is capable of infecting man and all warm-blooded animals. Cell-mediated immunity is vital in mounting protective responses against T. gondii infection. Recent studies have shown that T-helper (Th) 17 responses may play a key role in parasite control. In this current study, we constructed a DNA vaccine encoding T. gondii ROP13 in a pcDNA vector. Groups of BALB/c mice were immunized intramuscularly with pcROP13 or controls and challenged with the RH strain of T. gondii. The results showed that immunization with pcROP13 could elicit an antibody response against T. gondii. The expression of the canonical Th17 cytokines, interleukin (IL)-17 and IL-22, were significantly increased after immunization with pcROP13 compared with control groups ( p < 0.05). Furthermore, vaccination resulted in a significant decrease in parasite load ( p < 0.05). The induction of Th17 related cytokines, using a ROP13 DNA vaccine, against T. gondii should be considered as a potential vaccine approach for the control of toxoplasmosis.

IL-17 contributes to cardiac fibrosis following experimental autoimmune myocarditis by a PKCß/Erk1/2/NF-κB-dependent signaling pathway.

Myocarditis is a common clinical cardiovascular disease, and some patients progress to dilated cardiomyopathy (DCM) with chronic heart failure. Common viral infections are the most frequent cause of myocarditis, but other pathogens and autoimmune diseases have also been implicated. T(h)17 cells are novel IL-17-producing effector T helper cells that play an important role in the development of autoimmune myocarditis. Furthermore, IL-17 is also important in post-myocarditis cardiac remodeling and progression to DCM. However, the mechanisms whereby IL-17 and IL-17-producing cells promote the progression of cardiac fibrosis remain unclear. We therefore investigated whether IL-17 directly induced cardiac fibrosis in experimental autoimmune myocarditis (EAM) and explored the possible molecular mechanisms. The EAM model was induced and serum IL-17 level was detected by ELISA; western blot, immunofluorescence and sirius red staining were used to analyze the collagen expression. PCR was used to assay the IL-17RA and IL-17RC. The results indicated that IL-17 induced cardiac fibrosis both in vitro and in vivo. The protein kinase C (PKC)ß/Erk1/2/NF-κB (Nuclear Factor κappa B) pathway was involved in the development of myocardial fibrosis and IL-17 contributed to cardiac fibrosis following EAM via this pathway. These results provide the first direct evidence for the involvement of the PKCß/Erk1/2/NF-κB signaling pathway in IL-17-induced myocardial fibrosis.

HMGB1 blockade attenuates experimental autoimmune myocarditis and suppresses Th17-cell expansion.

High-mobility group box 1 (HMGB1), a non-histone nuclear protein, has been implicated in cardiovascular diseases. Dilated cardiomyopathy (DCM), one of the leading causes of heart failure, is often caused by coxsackievirus B3-triggered myocarditis and promoted by the post-infectious autoimmune process. Th17 cells, a novel CD4(+) T subset, may be important in the pathogenesis of autoimmune myocarditis. In the present study, we attempted to block HMGB1 function with a monoclonal antibody specific for HMGB1 B box and investigated the effects of the blockade on Th17 cells and experimental autoimmune myocarditis (EAM). After induction of EAM, HMGB1 protein levels were significantly elevated both in the heart and blood. Administration of an anti-HMGB1 B box mAb attenuated cardiac pathological changes and reduced the number of infiltrating inflammatory cells in the heart during EAM. These protective effects of HMGB1 blockade correlated with a reduced number of Th17 cells in local tissues and lower levels of IL-17 in the serum. Furthermore, in vitro, studies demonstrated that HMGB1 promoted Th17-cell expansion. Therefore, we speculate that HMGB1 blockade ameliorates cardiac pathological changes in EAM by suppressing Th17 cells.

Increased frequency of Th17 cells in the peripheral blood of children infected with enterovirus 71.

Enterovirus 71 (EV71) affects the health of young children globally causing severe neurologic diseases. The relationship between EV71 infection and T helper type 17 (Th17) has not been described, although this new Th subset or interleukin-17 (IL-17) has been reported to be associated with other viral infections. The purpose of the current study was to describe the immune profile involving Th17 cells, neutrophils, and related factors and to speculate on the possible immunopathogenesis of EV71 infections. Flow cytometry and an automatic blood cell counter were used to analyze circulating Th17 cells and count neutrophils, respectively. Expression of acid-related orphan nuclear receptor gamma t (ROR γt) was evaluated by reverse-transcriptional PCR, and enzyme linked immunosorbent assays (ELISAs) were used for detecting concentrations of IL-17, IL-23, and IFN-γ. The results showed that the frequencies of Th17 cells (1.47 ± 0.87%) and the number of neutrophils (7.4 ± 4.1 × 10(9) /L) in peripheral blood samples from children infected with EV71 were significantly higher compared to controls. In addition, there was a statistically higher expression of ROR γt in peripheral blood mononuclear cells (PBMCs) and elevated concentrations of IL-17 and IL-23 in sera, but lower IFN-γ production during EV71 infections. The findings suggest that Th17 cells are mediators during the immunologic process.

Herbaspirillum species: a potential pathogenic bacteria isolated from acute lymphoblastic leukemia patient.

Herbaspirillum species, colonized the plant rhizosphere, also called rhizobacteria, are plant growth-promoting bacteria. Recently we isolated Herbaspirillum from blood cultures of acute lymphoblastic leukemia (ALL) and identified by PCR and gene sequencing. Herbaspirillum may be a potential pathogenic bacteria. Although the exact role that these species play in ALL patients is unknown, their differentiation from other species has serious implications for clinical care and patient well-being.

Effect of IL-36γ expression on chronic periodontitis with an increase in NF-KB signaling pathway.

Background: An inappropriate inflammatory response is the cause of many common diseases, especially periodontitis. Considering that no studies have been carried out to investigate the effect of IL-36γ on chronic periodontitis, this study aimed to investigate the inflammatory mechanism of IL-36γ by stimulating macrophage cells using NF-KB pathway. Methods: This experimental study was performed on 50 healthy individuals and 50 subjects with chronic periodontitis. In this study, macrophage cells were extracted first, and then RNA was isolated from all the samples using TRIzol method. Subsequently, the rate of IL-36γ gene expression was analyzed and compared using real-time PCR technique. Additionally, immunofluorescence (IF) technique was used to investigate the rate of inflammation. The rate of NF-Kb expression was also measured via western blot technique. Finally, statistical analysis of the samples was carried out using appropriate statistical methods with SPSS 17. Results: The results showed that the rate of IL-36γ expression in subjects with periodontitis was higher compared to healthy subjects (P<0.05). Moreover, the results showed that following treatment of cells with TLR4, the rate of IL36γ expression increased significantly, especially during the 12-hour period after treatment. Conclusion: This indicates that after stimulating the TLR pathways, the rate of IL-36γ expression will probably increase.

Metabolic syndrome mediates inflammatory and oxidative stress responses in patients with recurrent pregnancy loss.

Recurrent pregnancy loss (RPL) is defined as three or more consecutive pregnancy losses prior to the 20th week of gestation. Exaggerated maternal immune response and oxidative stress status have been proposed as one of the main underlying mechanisms for RPL. The aim of this study was to evaluate the role of inflammatory pathway and oxidative stress imbalance in RPL patients with or without metabolic syndrome (MetS). 21 and 28 RPL patients with (RPL-MS) and without (RPL-NMS) metabolic syndrome were enrolled in this clinical study. 42 healthy women also were considered as the control group. The levels of IL1ß, IL6, IL17, TNFα, CCL2, CXCL8 were evaluated by ELISA method. Additionally, the oxidative stress biomarkers including TAS, TOS, NO, CAT, SOD, AOPP, MPO were analyzed by spectrophotometry. The expression levels of IL1ß, IL6, IL17, TNFα, CCL2, CXCL8, NFĸB, AP1, miR-21, miR-146-a, miR-223 were also assessed by real time PCR. The frequency of Th17 and T-reg cells was also measured by flow cytometry. Significant increase in the expression levels of IL1ß, IL6, IL17, TNFα, CCL2, CXCL8, NFĸB, AP1 and miR-21 was observed in RPL-MS patients. Furthermore, significant decreased expression levels of FoxP3, miR-146-a and miR-223 was also observed in RPL-MS group. The levels of IL1ß, IL6, IL17, TNFα, CCL2, CXCL8, NO, MPO and TOS were found to be higher in RPL-MS group compared to the RPL-NMS and healthy controls. In contrast, the level of CAT and SOD in RPL-MS patients was decreased. The frequency of Th17 and Treg cells was also higher and lower in RPL-MS patients compared to the other groups, respectively. Our results support the concept that subclinical inflammatory state, oxidative stress and metabolic syndrome play a crucial role in the etiopathogenesis of RPL assisting clinicians for pregnancy consequences prediction.

Evaluation of the effect of IL-36γ expression on chronic periodontitis by enhancing the MAPK and TLR4 signaling pathways: A basic research.

Background. Periodontitis is an infectious and inflammatory disease of the supporting tissues of the tooth caused by specific microorganisms or a group of microorganisms and, if not treated, leads to progressive degradation of the supporting tissues and subsequent loss of the teeth affected. The aim of this study was to evaluate the effects of IL-36γ on periodontitis by enhancing the TLR4 and MAPK signaling pathways. Methods. In this pilot study, 50 patients with generalized moderate-to-severe chronic periodontitis and 50 individuals with healthy periodontium, who were candidates for crown lengthening (CL), were selected based on inclusion criteria. The tissue samples were taken during pocket depth surgery (for the test group) and CL surgery (for the control group). The macrophage cells of the inflammatory tissues were extracted and stimulated by TLR4 proteins in a time-dependent manner; then IL-36γ levels in macrophages were investigated. Data were analyzed using descriptive statistics (means ± standard deviations and frequency percentages). Repeat measurement test was used to compare IL36γ expression in MAPK and TLR4 pathways at different time intervals. ANCOVA was used to compare IL36γ expression at different time intervals between the two pathways. Statistical analysis was performed using SPSS 17 at a significance level of P<0.05. Results. The results of the current study showed a significant relation between TLR4 and IL-36γ (P<0.001); in tissues with generalized moderate-to-severe chronic periodontitis, there was a significant relation between the condition and IL-36γ (P<0.0001). This study also showed that TLR4 and MAPK levels increased in the presence of IL-36γ. Conclusion. According to the present study, it was concluded that IL-36γ concentrations increased in periodontitis, which could trigger MAPK and TLR4 pathways.