Toll-like receptor 3 (TLR3) and the development of type 1 diabetes mellitus

Type 1 diabetes mellitus (T1DM) is a chronic, progressive autoimmune disease characterized by metabolic decompensation often leading to dehydration and ketoacidosis. Viral agents seem to play an important role in triggering the autoimmune destruction that leads to the development of T1DM. Among several viral strains investigated so far, the enterovirus family has been consistently associated with the onset of T1DM in humans. One of the mediators of viral damage is the double-stranded RNA (dsRNA) generated during replication and transcription of viral RNA and DNA. The Toll-like receptor 3 (TLR3) gene codes for an endoplasmic receptor of the pattern-recognition receptors (PRRs) family that recognizes dsRNA, plays an important role in the innate immune response triggered by viral infection. Binding of dsRNA to the TLR3 triggers the release of proinflammatory cytokines, such as interferons, which exhibit potent antiviral action; thus, protecting uninfected cells and inducing apoptosis of infected ones. Therefore, the TLR3 gene is a good candidate for the development of T1DM. Within this context, the objective of the present review was to address the role of the TLR3 gene in the development of T1DM. Arch Endocrinol Metab. 2015;59(1):4-12.

Role of inducible nitric oxide synthase on the development of virus-associated asthma exacerbation which is dependent on Th1 and Th17 cell responses

Asthma is characterized by airway inflammation induced by immune dysfunction to inhaled antigens. Although respiratory viral infections are the most common cause of asthma exacerbation, immunologic mechanisms underlying virus-associated asthma exacerbation are controversial. Clinical evidence indicates that nitric oxide (NO) levels in exhaled air are increased in exacerbated asthma patients compared to stable patients. Here, we evaluated the immunologic mechanisms and the role of NO synthases (NOSs) in the development of virus-associated asthma exacerbation. A murine model of virus-associated asthma exacerbation was established using intranasal challenge with ovalbumin (OVA) plus dsRNA for 4 weeks in mice sensitized with OVA plus dsRNA. Lung infiltration of inflammatory cells, especially neutrophils, was increased by repeated challenge with OVA plus dsRNA, as compared to OVA alone. The neutrophilic inflammation enhanced by dsRNA was partly abolished in the absence of IFN-gamma or IL-17 gene expression, whereas unaffected in the absence of IL-13. In terms of the roles of NOSs, dsRNA-enhanced neutrophilic inflammation was significantly decreased in inducible NOS (iNOS)-deficient mice compared to wild type controls; in addition, this phenotype was inhibited by treatment with a non-specific NOS inhibitor (L-NAME) or an specific inhibitor (1400 W), but not with a specific endothelial NOS inhibitor (AP-CAV peptide). Taken together, these findings suggest that iNOS pathway is important in the development of virus-associated exacerbation of neutrophilic inflammation, which is dependent on both Th1 and Th17 cell responses.

Mechanisms and roles of the RNA-based gene silencing

RNA silencing is a remarkable type of gene regulation. This process has been found to occur in many different organisms such as plants (co-suppression), fungi (quelling), and animals (RNA interference; RNAi). Double-stranded RNA (dsRNA) is a potent trigger in RNA silencing mechanisms operating in a wide range of organisms. This mechanism recognizes dsRNA and processes them into small 21-25nt RNAs (smRNAs). Small RNAs can guide post-transcriptional degradation of complementary messenger RNAs and in plants, transcriptional gene silencing is occurred by methylation of homologous DNA sequences. In plants, it serves as an antiviral defense, and many plant viruses encode suppressors of silencing such as helper component-proteinase of potyviruses (HC-Pro) and the p25 protein encoded by potato virus X (PVX). HC-Pro acts by preventing accumulation of smRNAs that provide specificity determinant for homologous RNA degradation, but p25 viral protein acts by targeting the mobile silencing signal. The encouraging view is that RNA silencing is part of a sophisticated network of interconnected pathways for cellular defense and development and that it may become a powerful tool to manipulate gene expression experimentally.

Acridine orange metachromasia in the cytoplasm of simian rotavirus (SA-11)-infected MA-104 cell cultures

Acridine orange metachromasia was used to determine the distribution of simian rotavirus double-stranded RNA in cultured MA-104 cells 0 to 72 h post-infection. Correlations were made among time of detection and amount of viral antigens, virus yield and the ultrastructural aspects of infected cells. RNAase-resistant cytoplasmic metachromasia appeared 48 h post-infection, 36 h after the initial detection of viral antigens or infectious virions and 24 h after the appearance of the cytopathic effect. Acridine orange staining is thus useful for monitoring the progress of rotaviral infection in cell cultures due to its simplicity and low cost, in spite of its lower sensitivity compared to other techniques evaluated