Association of levels of interleukin 17 and T-helper 17 count with symptom severity and etiology of chronic heart failure: a case-control study

Aim To assess the association between the levels of interleukin 17 (IL-17) and T-helper 17 count and symptom severity and etiology of chronic heart failure. Methods This single-center prospective case-control study, conducted from December 1, 2015 to January 1, 2017 in Tehran Heart Center, evaluated gene expression of IL-17, relative count of (CD4+IL17+) Th17 cells and CD4+ helper T-cells in peripheral blood mononuclear cells of 42 patients with CHF and 42 matched controls. A multiple regression model assessed the predictors of peripheral IL-17 expression and Th17 count in patients with CHF. Results IL-17 expression was increased in patients with CHF, both at baseline and after stimulation. IL-17 and Th17 counts were higher in patients with advanced New York Heart Association (NYHA) functional class (class IV) than in controls and patients with class I. Th17 cell population expanded in patients with CHF, more prominently in patients with class IV than in controls and patients with class I, regardless of the ischemic or non-ischemic CHF origin. Multiple regression model showed that NYHA was the only meaningful predictor of IL-17 levels and Th17 count. Conclusion We demonstrated the lymphocytic origin of IL-17 production in advanced CHF and the ability of disease severity to predict IL-17 levels

Challenging TaqMan probe-based real-time PCR and loop-mediated isothermal amplification (LAMP): the two sensitive molecular techniques for the detection of toxoplasmosis, a potentially dangerous opportunistic infection in immunocompromised patients

Due to defects and drawbacks of most conventional diagnostic methods including serology for the diagnosis of toxoplasmosis as a dangerous opportunistic infection in immunocompromised individuals, the accurate, rapid, and sensitive detection of infection in such patients is essential. In this study, the TaqMan probe-based real-time PCR and, a relatively new nucleic acid amplification method, the loop-mediated isothermal amplification (LAMP) technique was compared based on the repetitive elements (RE) sequence to detect Toxoplasma gondii (T. gondii) DNA in blood samples of immunocompromised individuals. During this study, 119 blood samples from immunocompromised cancer patients with renal failure, undergoing dialysis were studied. After DNA extraction from blood samples using the salt extraction method, the molecular techniques of TaqMan probe-based real-time PCR and LAMP were used to investigate the contamination of the samples with T. gondii, based on the 529 bp (RE) sequence of T. gondii. The analytical sensitivity of LAMP and real-time PCR was evaluated by duplicating the five-step serial dilutions of T. gondii tachyzoites from 0.25 to 5x10(5) spiked tachyzoites per milliliter of the Toxoplasma seronegative blood sample. The extracted DNA from other parasites and human chromosomal DNA were used to determine the specificity of the molecular methods. The obtained results were analyzed using Kappa statistical test and SPSS22 software. Out of 119 studied samples, 7 (5.8%) and 5 (4.2%) samples were positive for Toxoplasma by TaqMan probe-based real-time PCR and LAMP, respectively. The limits of detection of TaqMan probe-based real-time PCR and RE-LAMP in negative serum samples were one and five tachyzoites (CT 38), respectively. Both real-time PCR and LAMP methods were 100% specific for Toxoplasma detection. Positive results were obtained only with T. gondii DNA, while other DNA samples were negative. The TaqMan probe-based real-time PCR based on the RE sequence showed higher sensitivity to T. gondii DNA detection in blood samples of cancer patients and serial dilutions of parasitic tachyzoites. The results show that TaqMan probe-based real-time PRC is a sensitive and specific method for the detection of toxoplasmosis in immunocompromised individuals, as well as the LAMP assay, which can be used as a suitable alternative diagnostic method for the detection of toxoplasmosis in such patients, without need the for any expensive equipment. Keywords:LAMP; TaqMan probe-based real-time PCR; RE gene; Toxoplasmosis; Immunocompromised patient

Increased Levels of IL-23 in Peripheral Blood Mononuclear Cells of Patients With Chronic Heart Failure

Chronic heart failure (CHF) is a complex clinical syndrome that represents the end stage of various cardiac diseases and is characterized by the inability of the heart to meet metabolic demands of the body. Many physiological systems are involved in this disease. In particular, the activation of the immune system has received considerable interest in the last decade. Evidence from both experimental and clinical trials indicates that inflammatory mediators are of importance in the pathogenesis and progression of chronic heart failure. Excessive pro-inflammatory cytokines induce contractile dysfunction, hypertrophy, and fibrosis and cell death in Cardiac myocyte. We examined the expression of IL-23 in PBMCs between CHF patients and healthy controls. In this report, we used real-time PCR assay to compare the relative expression of IL-23 in peripheral blood mononuclear cells (PBMC) from CHF patients with various heart diseases (n=42, EF<45%, range of New York Heart Association (NYHA) 1 to 4) and matched healthy control subjects (n=42).We also determined the IL-23 concentrations of cell culture supernatant of PBMCs with ELISA. A total of 42 patients with CHF, with 42 age and sex-matched control group subjects were enrolled in the present study. The culture supernatant levels of IL-23 in PBMC of CHF patients were significantly higher (133.95±108.99 pg/mL) than in the control group (83.43±76.2 pg/mL) (P<0.05). The gene expression of IL-23 was also markedly upregulated in PBMC from CHF patients in comparison with the control group, but it was not statically significant 80. These results demonstrate that in patients with CHF and especially those with severe CHF, expression of pro-inflammatory cytokines and levels of IL-23 cytokine is markedly increased in PBMC. These finding suggested that IL-23 may play an important role in the progression of CHF among these patients