miR-1224 Expression is Increased in Human Macrophage After Infection With Bacillus Calmette-Guerin (BCG)

Introduction: Tuberculosis remains a major threat to human health. Understanding the strategies mycobacterium takes to overcome immune defense is important to control the infection. miRNAs are master regulators of most pathways in the human body. Infection with mycobacterium impacts upon host metabolic pathways to obtain the nutrition for its intracellular survival. In this study, we aimed to investigate the effect of BCG infection on the expression of three miRNAs (miR-1224, -484 and -425), which have been previously demonstrated to be important in infection and metabolic pathways. Methods: Blood-derived monocyte derived macrophage (MDM) cultures were prepared and infected with BCG at a multiplicity of infection (MOI) =10 or left uninfected as a control. 72h post-infection, the cultured cells were subjected to RNA extraction, cDNA synthesis and real-time PCR. Expression levels miRNAs were normalized to the levels of U6 snRNA (Rnu6) using the 2–ΔΔCt method Results: Infection with BCG resulted in a highly significant increase in miR-1224 expression (24.4±3.8-fold induction) in human MDMs. The induction of miR-484 (1.8±0.3-fold increase) and of miR-425 (1.2±0.2-fold increase) was less increased compared to miR-1224. Discussion: Mycobacterium tolerate an hostile microenvironment by escaping from lysosomal degradation and providing a lipid-rich niche by trigger with and re-pattering host metabolism. These data highlight the potential importance of miR-1224 in the host metabolic response to TB infection and a possible role of miR-484 in the reprogramming of metabolic pathways in infected cells. Further work is required to determine whether these miRNAs may also be useful as biomarkers for the diagnosis or monitoring of treatment in TB patients. Conclusion: This study Highlighted the potential roles of miRNAs in host immunologic responses upon mycobacterium infection. Further work is required to determine whether these miRNAs may also be useful as biomarkers for the diagnosis or monitoring of treatment in TB patients

Clinical Significance and Different Expression of Dipeptidyl Peptidase IV and Procalcitonin in Mild and Severe COVID-19

Background: Coronavirus has become a global concern in 2019-20. The virus belongs to the coronavirus family, which has been able to infect many patients and victims around the world. The virus originated in the Chinese city of Wuhan, which eventually spread around the world and became a pandemic. Materials and Methods: A total of 60 Patients with severe (n=30) and mild (n=30) symptoms of COIVD-19 were included in this study. Peripheral blood samples were collected from the patients. Real-time PCR was used to compare the relative expression levels of Procalcitonin and dipeptidyl peptidase IV (DPPIV) in a patient with severe and mild Covid-19 infection. Results: Procalcitonin and dipeptidyl peptidase IV markers in the peripheral blood of patients with severe symptoms, were positive in 29 (96.60%) and 26 (86.60%), respectively (n=30); however, positive rates in the mild symptoms patients group were 27 (90%) and 25 (83.30%), respectively. There was a statistically significant difference between these two groups in terms of DDPIV and Procalcitonin (p<0.001). Conclusion: Procalcitonin and DPPIV increase in patients with COVID-19 infection, significantly higher in the patients with more severe clinical symptoms than those with milder ones. More studies will be needed to verify the reliability of the current findings. Keywords: Procalcitonin, DPPIV, Severe symptoms, Mild symptoms, COVID-1

Immunophenotype and function of circulating myeloid derived suppressor cells in COVID-19 patients

The pathogenesis of coronavirus disease 2019 (COVID-19) is not fully elucidated. COVID-19 is due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) which causes severe illness and death in some people by causing immune dysregulation and blood T cell depletion. Increased numbers of myeloid-derived suppressor cells (MDSCs) play a diverse role in the pathogenesis of many infections and cancers but their function in COVID-19 remains unclear. To evaluate the function of MDSCs in relation with the severity of COVID-19. 26 PCR-confirmed COVID-19 patients including 12 moderate and 14 severe patients along with 11 healthy age- and sex-matched controls were enrolled. 10 ml whole blood was harvested for cell isolation, immunophenotyping and stimulation. The immunophenotype of MDSCs by flow cytometry and T cells proliferation in the presence of MDSCs was evaluated. Serum TGF-β was assessed by ELISA. High percentages of M-MDSCs in males and of P-MDSCs in female patients were found in severe and moderate affected patients. Isolated MDSCs of COVID-19 patients suppressed the proliferation and intracellular levels of IFN-γ in T cells despite significant suppression of T regulatory cells but up-regulation of precursor regulatory T cells. Serum analysis shows increased levels of TGF-β in severe patients compared to moderate and control subjects (HC) (P = 0.003, P < 0.0001, respectively). The frequency of MDSCs in blood shows higher frequency among both moderate and severe patients and may be considered as a predictive factor for disease severity. MDSCs may suppress T cell proliferation by releasing TGF-β

Susceptibility to mycobacterial disease due to mutations in IL-12Rβ1 in three Iranian patients

In the last decade, autosomal recessive interleukin-12 receptor β1 (IL-12Rβ1) deficiency, the most common cause of Mendelian susceptibility to mycobacterial disease (MSMD), has been diagnosed in a few children and adults with severe tuberculosis in Iran. Here, we report three cases referred to the Immunology, Asthma and Allergy ward at the National Research Institute of Tuberculosis and Lung Diseases (NRITLD) at Masih Daneshvari Hospital from 2012 to 2017 with Mycobacterium tuberculosis and non-tuberculous mycobacteria infections due to defects in IL-12Rβ1 but with different clinical manifestations. All three were homozygous for either an IL-12Rβ1 missense or nonsense mutation that caused the IL-12Rβ1 protein not to be expressed on the cell membrane and completely abolished the cellular response to recombinant IL-12. Our findings suggest that the presence of IL-12Rβ1 deficiency should be determined in children with mycobacterial infections at least in countries with a high prevalence of parental consanguinity and in areas endemic for TB like Iran

The Recent-Transmission of Mycobacterium tuberculosis Strains among Iranian and Afghan Relapse Cases: a DNA-fingerprinting using RFLP and spoligotyping

<p>Abstract</p> <p>Background</p> <p>Relapse of tuberculosis (TB) may develop as the result of reactivation of the endogenous primary infection, or as a result of a exogenous reinfection. This survey evaluated the rate of reactivation versus recent transmission among Iranian and Afghan relapse cases.</p> <p>Methods</p> <p>The sputum specimens were digested, examined microscopically for acid-fast bacilli, and inoculated into Löwenstein-Jensen slants by standard procedures. Thereafter, the susceptibility and identification tests were performed on culture positive specimens. Subsequently, the strains that were identified as <it>Mycobacterium tuberculosis </it>(258 isolates) were subjected to IS6110 restriction fragment length polymorphism (RFLP) and spoligotyping. Additional patient's information was collected for further epidemiological analysis. Patients whose isolates had identical genotyping patterns were considered a cluster with recent transmission episode.</p> <p>Results</p> <p>Out of 258 available isolates, 72(28%) had multi-drug resistant (MDR-TB) in ratio and 42 (16.2%) had other resistant. Notably, 38 of MDR-TB cases (52%) were isolated from Afghan patients. By IS6110-RFLP typing method, 65 patients (25%) were clustered in 29 clusters. In cluster cases, the intra-community transmissions between Iranian and Afghan patients were 41%. All MDR-TB patients in clusters had either Haarlem I or Beijing characteristic. The risk factors like sex, family history, close contact, living condition, PPD test result and site of TB infection were not associated with clustering. Although, the MDR-TB strains were more frequent in non-cluster cases (31%) than cluster one(18%) (P < 0.05). Majority of <it>M. tuberculosis </it>strains isolated from non-cluster cases were belong to EAI3 (51; 30%) and CASI(32;18.6%) superfamilies.</p> <p>Conclusion</p> <p>During the studied period, reactivation of a previous infection remain the more probable cause of recurrence. Although, the evidence of intra- community transmission between Iranian and Afghan TB cases, highlighted the impact of afghan immigrants in national tuberculosis control program (NTP) of Iran.</p

Water-pipe smoke condensate increases the internalization of Mycobacterium Bovis of type II alveolar epithelial cells (A549)

Background: Tuberculosis (TB) is a major global health problem, and there is an association between tobacco smoke and TB. Water pipe smoking has become an increasing problem not only in Middle Eastern countries but also globally because users consider it as safer than cigarettes. The presence of high levels of toxic substances in water-pipe smoke may be a predisposing factor that enhances the incidence of pulmonary disorders. For example, uncontrolled macropinocytosis in alveolar epithelial cells following exposure to water-pipe smoke may predispose subjects to pulmonary infection. Here, we studied the effects of water-pipe condense (WPC) on the internalization of Mycobacterium Bovis BCG by macropinocytosis in the alveolar epithelial cell line A549. Methods: A549 cells were exposed to WPC (4 mg/ml) for 24, 48, 72 and 96 h. Cell viability was studied using the methyl thiazolyldipenyl-tetrazolium bromide (MTT) reduction assay and proliferation by bromodeoxyUridine (BrdU) incorporation. Cells were exposed to FITC-Dextran (1 mg/ml) (as a control) and FITC-BCG (MOI = 10) for 20 min at 37 ° Cbeforecellswere collected and the uptake of BCG-FITC determined by flow cytometry. Similar experiments were performed at 4 ° Casacontrol . The Rho-associated protein kinase (ROCK) inhibitor Y-27632 (1 μ M) was used to assess the mechanism by which WPC enhanced BCG uptake. Results: WPC (4 mg/ml) increased the uptake of BCG-FITC after 72 (1.3 ± 0.1 fold, p < 0.05) and 96 (1.4 ± 0.05 fold, p < 0.05) hours. No effect on BCG-FITC uptake was observed at 24 or 48 h. WPC also significantly increased the uptake of FITC-Dextran (2.9 ± 0.3 fold, p < 0.05) after 24 h. WPC significantly decreased cell viability after 24 (84 ± 2%, p < 0.05), 48 (78±, 3%, p < 0.05), 72 (64 ± 2%, p < 0.05) and 96 h (45 ± 2%, p < 0.05). Y-27632 completely attenuated the increased uptake of BCG by WPC. Cell proliferation showed a decreasing trend in a time-dependent manner with WPC exposure. Conclusion: WPC exposure increased epithelial cell endocytosis activity and death as well as enhancing their capacity for macropinocytosis. Our in vitro data indicates possible harmful effects of WPC on the ability of lung epithelial cells to phagocytose mycobacterium

Susceptibility to tuberculosis among pulmonary tuberculosis patients: p2X7 and TNF-α gene polymorphisms

Introduction: This study was conducted to assess the rate of human P2X7 and TNF-α gene polymorphisms among Iranian pulmonary tuberculosis (TB) cases. P2X7 gene is highly polymorphic, and it is up-regulated by an inflammatory cytokine. Recently, a correlation between the expression of the P2X7 receptor and the elevated levels of TNF-α have been shown, but the apparent clinical correlation could not be demonstrated. Methodology: This study included 80 confirmed pulmonary tuberculosis (PTB) cases. Fifty control subjects were selected from the TB patients (clinical and laboratory) who had positive tuberculin tests (10–15 mm) and showed no sign of diseases (laboratory, radiological and clinical parameter). Single nucleotide polymorphisms (SNPs) in P2X7 (+1513, −762) and TNF-α (at −238, −308, −244, −857, −863) genes were assessed using PCR-RFLP and allele-specific PCR. Thereafter, haplotype and diplotype variability were compared and analyzed. Results: For 1513 loci, the heterozygosity was higher in patients (35%; 44.3%) than control subjects (12%; 24%) [(p = 0.026) ORS; 2.45 CI 95% (1.13–5.33)]. For −762 loci, the frequency of mutant alleles between patients and controls were not statistically significant. No statistical difference was observed in allele frequencies of TNF −308 and −857. However, the frequency of −238 allele were more in TB cases (72.1%) (p = 0.000)[ORs: 5.85(2.70–12.64). Data analysis showed more frequencies of haplotypes, i.e., TGGA-CA and CGGA-TA in patients (21.5%; 14.6%) than the control group (2.0%; 6.0%), respectively. Additionally, the diplotype “CCGGGGGG-CCAA” was significantly associated with susceptibility to PTB (1.9 [0.08–48.3]). Conclusions: In the studied population, polymorphisms in P2X7 (1513) and TNF-α (−238) gene was associated with the risk of developing PTB. Additionally, distribution of haplotype and diplotype variables did appear to be more specific than SNPs