RESUMO
Rotavirus is the main cause of acute diarrhea in children up to five years of age. In this regard, probiotics are commonly used to treat or prevent gastroenteritis including viral infections. The anti-rotavirus effect of Bifidobacterium longum and Chlorella sorokiniana, by reducing viral infectivity and improving IFN-type I response, has been previously reported. The present study aimed to study the effect of B. longum and/or C. sorokiniana on modulating the antiviral cellular immune response mediated by IFN-γ, IL-10, SOCS3, STAT1, and STAT2 genes in rotavirus-infected cells. To determine the mRNA relative expression of these genes, HT-29 cells were treated with B. longum and C. sorokiniana alone or in combination, followed by rotavirus infection. In addition, infected cells were treated with B. longum and/or C. sorokiniana. Cellular RNA was purified, used for cDNA synthesis, and amplified by qPCR. Our results demonstrated that the combination of B. longum and C. sorokiniana stimulates the antiviral cellular immune response by upregulating IFN-γ and may block pro-inflammatory cytokines by upregulating IL-10 and SOCS3. The results of our study indicated that B. longum, C. sorokiniana, or their combination improve antiviral cellular immune response and might modulate pro-inflammatory responses.
Assuntos
Bifidobacterium longum , Chlorella , Interferon gama , Interleucina-10 , Probióticos , Infecções por Rotavirus , Proteína 3 Supressora da Sinalização de Citocinas , Humanos , Células HT29 , Interferon gama/metabolismo , Interleucina-10/metabolismo , Probióticos/farmacologia , Rotavirus/fisiologia , Infecções por Rotavirus/imunologia , Infecções por Rotavirus/virologia , Fator de Transcrição STAT1/metabolismo , Proteína 3 Supressora da Sinalização de Citocinas/metabolismoRESUMO
Hypoxia-inducible factor -1 (HIF-1) is a transcriptional factor that regulates the expression of several glycolytic genes. The white spot syndrome virus (WSSV) induces a shift in glycolysis that favors viral replication in white shrimp Litopenaeus vannamei. HIF-1 is related to the pathogenesis of the WSSV infection through the induction of metabolic changes in infected white shrimp. Although the WSSV infection is associated with metabolic changes, the role of HIF-1 on key glycolytic genes during the WSSV infection has not been examined. In this work, we evaluated the effect of HIF-1α silencing on expression and activity of glycolytic enzymes (Hexokinase-HK, phosphofructokinase-PFK and pyruvate kinase-PK) along with the glucose transporter 1 (Glut1), regulatory enzymes (glucose-6-phosphate dehydrogenase-G6PDH and pyruvate dehydrogenase-PDH), and metabolic intermediates of glycolysis (glucose-6-phosphate-G6P and pyruvate). The expression of Glut1 increased in each tissue evaluated after WSSV infection, while HK, PFK and PK gene expression and enzyme activities increased in a tissue-specific manner. G6PDH activity increased during WSSV infection, and its substrate G6P decreased, while PDH activity decreased and its substrate pyruvate increased. Silencing of HIF-1α blocked the WSSV-induced Glut1 and glycolytic genes upregulation and enzyme activity in a tissue-specific manner. We conclude that HIF-1 regulates the WSSV-induced glycolysis through induction of glycolytic genes contributing to glucose metabolism in tissues of infected shrimp. Also, the inhibition, and activation of regulatory genes are likely to decrease the availability of the raw materials essential for WSSV replication and increase oxidative metabolism.
Assuntos
Glicólise/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/imunologia , Penaeidae/genética , Penaeidae/imunologia , Vírus da Síndrome da Mancha Branca 1/fisiologia , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Regulação da Expressão Gênica/imunologia , Inativação GênicaRESUMO
Annual mortality rates due to infectious diarrhea are about 2.2 million; children are the most vulnerable age group to severe gastroenteritis, representing group A rotaviruses as the main cause of disease. One of the main factors of rotavirus pathogenesis is the NSP4 protein, which has been characterized as a viral toxin involved in triggering several cellular responses leading to diarrhea. Furthermore, the rotavirus protein NSP1 has been associated with interferon production inhibition by inducing the degradation of interferon regulatory factors IRF3, IRF5, and IRF7. On the other hand, probiotics such as Bifidobacterium and Lactobacillus species in combination with prebiotics such as inulin, HMO, scGOS, lcFOS have been associated with improved generalized antiviral response and anti-rotavirus effect by the reduction of rotavirus infectivity and viral shedding, decreased expression of NSP4 and increased levels of specific anti-rotavirus IgAs. Moreover, these probiotics and prebiotics have been related to shorter duration and severity of rotavirus diarrhea, to the prevention of infection and reduced incidence of reinfections. In this review we will discuss in detail about the rotavirus pathogenesis and immunity, and how probiotics such as Lactobacillus and Bifidobacterium species in combination with prebiotics have been associated with the prevention or modulation of rotavirus severe gastroenteritis.
Assuntos
Antivirais/uso terapêutico , Bifidobacterium/fisiologia , Gastroenterite/terapia , Lactobacillus/fisiologia , Prebióticos/estatística & dados numéricos , Probióticos/uso terapêutico , Infecções por Rotavirus/terapia , Rotavirus/efeitos dos fármacos , Criança , Diarreia/terapia , Diarreia/virologia , Feminino , Gastroenterite/virologia , Glicoproteínas/metabolismo , Humanos , Lactente , Inulina/uso terapêutico , Rotavirus/imunologia , Toxinas Biológicas/metabolismo , Proteínas não Estruturais Virais/metabolismoRESUMO
BACKGROUND AND OBJECTIVES: Enteric viral infections are among the leading causes of gastroenteritis in children up to five years of age worldwide. This study was aimed to determine the disease severity, incidence, and molecular genotyping of rotaviruses, noroviruses, astroviruses, and enteric adenoviruses as gastroenteritis agents among children up to five years old. MATERIALS AND METHODS: Gastroenteritis severity was determined by using the Ruuska and Vesikari score, whereas the incidence of enteric infections and their genotyping were determined by reverse transcription-polymerase chain reaction (RT-PCR) and sequence analysis. RESULTS: Rotaviruses were observed to possess the highest incidence with 10% (18/179) of the cases positives; nevertheless, noroviruses had the highest severe gastroenteritis score (13⯱â¯3 points). Results indicated that 56% (10/18) of the detected rotavirus strains were genotype G12P[8], 50% (4/8) of noroviruses were GII.4 and 25% (2/8) were genotype GI.8. Out of the sapovirus positive samples, 30% (2/6) were genotyped as GI·I and GII·I. Sixty percent of the astrovirus strains (3/5) were genotype HAstV-2, and 20% (1/5) were genotype HAstV-6. Additionally, one of the adenovirus strains was identified as human mastadenovirus C type 6 specie. CONCLUSIONS: The diarrhea severity reduction in children provides evidence that the rotavirus vaccination program in the northwest of Mexico has been successful, even among children infected by the rotavirus emergent strain G12, however, norovirus resulted as the leading severe gastroenteritis-causing agent in children with rotavirus vaccine.