RESUMO
Rabbit hemorrhagic disease virus (RHDV) causes a highly contagious disease in rabbits that is associated with high mortality. Because of the lack of a suitable cell culture system for RHDV, its pathogenic mechanism and replication remain unclear. This study found that the expression level of host protein rabbit hemoglobin subunit beta (HBB) was significantly downregulated in RHDV-infected cells. To investigate the role of HBB in RHDV replication, small interfering RNAs for HBB and HBB eukaryotic expression plasmids were used to change the expression level of HBB in RK-13 cells and the results showed that the RHDV replication level was negatively correlated with the expression level of HBB. It was also verified that HBB inhibited RHDV replication using constructed HBB stable overexpression cell lines and HBB knockout cell lines. The interaction of HBB with viral capsid protein VP60, replicase RdRp, and VPg protein was confirmed, as was the activation of the expression of interferon γ by HBB. The results of this study indicated that HBB may be an important host protein in host resistance to RHDV infection.
Assuntos
Infecções por Caliciviridae/veterinária , Proteínas do Capsídeo/metabolismo , Subunidades de Hemoglobina/metabolismo , Vírus da Doença Hemorrágica de Coelhos/química , Vírus da Doença Hemorrágica de Coelhos/metabolismo , Proteínas Virais/metabolismo , Proteínas Estruturais Virais/metabolismo , Replicação Viral , Animais , Proteínas do Capsídeo/genética , Linhagem Celular , Feminino , Subunidades de Hemoglobina/genética , Subunidades de Hemoglobina/imunologia , Vírus da Doença Hemorrágica de Coelhos/genética , Vírus da Doença Hemorrágica de Coelhos/fisiologia , Interferon gama/imunologia , Coelhos , Proteínas Virais/genéticaRESUMO
BACKGROUND: Rhinitis comprises several diseases with varying causes and different clinical manifestations and pathological features, but treated as a single clinical disorder. As heterogeneous disease, proper differential diagnosis is useful to delineate appropriate therapeutic intervention. Comparative proteomic investigation was aimed to provide information for specific differentially expressed proteins in rhino pathologic state, that could be used for diagnostic purpose and therapeutic monitoring. METHODS: Proteins extracted from nasal mucosa cells of patients with different features of rhinitis and from control subjects, were separated by 2-DE. Proteins differentially expressed were identified by mass spectrometry (MS). RESULTS: Comparative proteomic analyses led to the identification of eighteen proteins differentially expressed in patients with rhinitis, mainly related to cell defense and innate and acquired immunity. From that, at least one protein can be a possible candidate as biomarker of disease.
Assuntos
Mucosa Nasal/imunologia , Mucosa Nasal/patologia , Rinite/genética , Rinite/imunologia , Adulto , Aldeído Desidrogenase/imunologia , Família Aldeído Desidrogenase 1 , Aldeído-Desidrogenase Mitocondrial/imunologia , Antígenos de Neoplasias/imunologia , Biomarcadores , Eletroforese em Gel Bidimensional , Eosinófilos/patologia , Feminino , Glutationa S-Transferase pi/imunologia , Glutationa Transferase/imunologia , Glicoproteínas/imunologia , Subunidades de Hemoglobina/imunologia , Humanos , Isoenzimas/imunologia , Masculino , Espectrometria de Massas , Mastócitos/patologia , Pessoa de Meia-Idade , Pólipos Nasais/imunologia , Pólipos Nasais/patologia , Neutrófilos/patologia , Peroxirredoxinas/imunologia , Fosfoproteínas/imunologia , Proteômica , Retinal Desidrogenase , Proteínas S100/imunologia , Proteínas de Ligação a Selênio/imunologia , Serpinas/imunologia , Albumina Sérica/imunologia , Tiorredoxinas/imunologiaRESUMO
Hemoglobin (Hb) is the major protein component of erythrocytes in animals with red blood, but it can serve additional functions beyond the transport of oxygen. In this study, we identified polymorphism in the blood clam Tegillarca granosa Hb (Tg-Hb) genes and investigated the association of this polymorphism with resistance/susceptibility to Vibrio parahaemolyticus. Analysis of the 540 sequences revealed 28 SNPs in the coding region of three Tg-Hbs, corresponding to about one SNP per 48 bp. Three SNPS: HbIIA-E2-146, HbIIB-E2-23, HbIIB-E2-121 showed a significant association with resistance/susceptibility to V. parahaemolyticus (P < 0.05). To further demonstrate that three significant SNPs of Tg-Hbs is associated with resistance of clams to V. parahaemolyticus, SNPs were genotyped in V. parahaemolyticus resistant strain clams and the wild base population from which this strain was derived. The results indicated that the nonsynonymous mutation T allele at HbIIA-E2-146 and A allele at HbIIB-E2-23 are associated with V. parahaemolyticus resistance in the blood clam, and its association with disease resistance may be due to its cause changes in amino acid sequences to a functional polymorphism. Together with previous bacterial challenge study, these results provides direct evidence that variation at HbIIA-E2-146 and HbIIB-E2-23 are associated with disease resistance in the blood clam, and these two polymorphic loci could be potential gene markers for the future molecular selection of strains that are resistant to diseases caused by V. parahaemolyticus.