RESUMO
BACKGROUND: Respiratory syncytial virus (RSV) infects infants and children, predisposing them to development of asthma during adulthood. Epithelial neuroendocrine phenotypes may be associated with development of asthma. This study hopes to ascertain if RSV infection promotes epithelial neuroendocrine phenotypes through the NODAL signaling pathway. METHODS: The GSE6802 data set was obtained from the GEO database, and the differential genes were analyzed using the R language. An in vitro model was constructed with RSV infected human respiratory epithelial cells, and then real-time qPCR and immunofluorescence were used to detect the expression of different epithelial biomarkers and airway neuropeptides. The acute and chronic infection model of RSV infection was established by intranasal injection of RSV into guinea pigs. Immunohistochemistry and Western blot were used to detect the expression of pulmonary neuroendocrine cells markers ENO2 and neuropeptides. RESULTS: The expression levels of ENO2, SP, CGRP, and NODAL/ACTRII were significantly higher in the RSV infection group than those of the control group, which were abrogated by siRNA-NODAL. In vivo, we found that the expression levels of ENO2, SP, and CGRP were significantly higher than that of the control group. CONCLUSION: RSV promotes epithelial neuroendocrine phenotypes through the NODAL signaling pathway.
Assuntos
Células Neuroendócrinas/metabolismo , Ligantes da Sinalização Nodal/metabolismo , Infecções por Vírus Respiratório Sincicial/fisiopatologia , Animais , Asma/metabolismo , Diferenciação Celular , Linhagem Celular , China , Bases de Dados Factuais , Bases de Dados Genéticas , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Células Epiteliais/virologia , Cobaias , Células HeLa , Humanos , Pulmão/metabolismo , Células Neuroendócrinas/virologia , Neuropeptídeos/metabolismo , Proteína Nodal/genética , Proteína Nodal/metabolismo , Proteína Nodal/fisiologia , Ligantes da Sinalização Nodal/genética , Fenótipo , Infecções por Vírus Respiratório Sincicial/metabolismo , Vírus Sinciciais Respiratórios/patogenicidade , Transdução de SinaisRESUMO
BACKGROUND: The aim of this study was to investigate the role of Notch-1 signaling through Notch-1 ligands on bronchial epithelial cells (BECs) in regulating the development of T helper 2 (Th2) lymphocytes after RSV infection. METHODS: Firstly, we analyzed the expression of cytokines and Notch-1 ligands in BECs by using real-time PCR. Then, RSV-infected BECs were co-cultured with CD4+ T cells in a transwell chamber for 48 h, and differentiation of T cells in the lower chamber was determined using flow cytometry and real-time PCR. JAG1 siRNA was then used to determine the effects of Jagged/Notch-1 signaling on the differentiation of Th2. An RSV-infected mouse model was also used to analyze the secretion of Th differentiation-associated cytokines in serum and lung tissues using ELISA, the histopathological changes using HE staining, and the expression of JAG1 and JAG2 in BECs. RESULTS: The results showed that RSV promoted the expression of Th2-type cytokines and Jagged-1 and inhibited the expression of Jagged-2 in normal BECs. RSV-infected BECs induced Th2 differentiation. In addition, JAG1 downregulation inhibited the differentiation of Th2 and promoted differentiation of Th1. In the RSV-infected mouse model, the RSV titer, inflammation decreased with time. IL-4 and IL-17 increased on day 28 and 60, while IFNγ increased on day 7 and 28. Moreover, the expression of Jagged-1 increased and that of Jagged-2 decreased in BECs, which was consistent with IL-4 production in lung tissues. CONCLUSION: Our data showed that BECs had the potential to promote the differentiation of Th2 lymphocytes through Jagged-1/Notch-1 signaling.
Assuntos
Brônquios/fisiologia , Proteína Jagged-1/fisiologia , Proteína Jagged-2/fisiologia , Receptor Notch1/fisiologia , Infecções por Vírus Respiratório Sincicial/imunologia , Transdução de Sinais/fisiologia , Células Th2/citologia , Animais , Brônquios/imunologia , Brônquios/patologia , Diferenciação Celular , Citocinas/biossíntese , Células Epiteliais/fisiologia , Células HeLa , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
BACKGROUND: Respiratory syncytial virus (RSV) infection is associated with serious lung disease in infants and immunocompromised individuals and is linked to development of asthma. Infection of RSV has been shown to induce Th lymphocyte differentiation. The present study was designed to determine the effects of RSV on the expression of Notch-1 and the related mechanisms on subsequent differentiation of Th lymphocytes. METHODS: A RSV-infected animal model was established and investigated at 7, 28 and 60 days post infection. Real-time qPCR and Western blot were used to observe the expression levels of Notch-1 in CD4+ T cells and its five ligands in lung tissues. The methylation levels of CpG islands in autoimmune regulator (AIRE) and Notch-1 promoters were analysed by time-of-flight mass spectrometry. The differentiation of Th lymphocytes was assayed by real-time qPCR. The distribution of JAG1 and DLL3 in the lung tissues were assayed by immunohistochemistry. The correlation between Th17 and DLL3 was analysed by simple correlation. RESULTS: The results showed that RSV promoted the expression and de-methylation of Notch-1 promoters in CD4+ T cells. Moreover, RSV infection promoted Th1 differentiation at day 7 and day 28; Th17 differentiation at day 7, day 28 and day 60; Th2 differentiation at day 28 and day 60. At the same time, RSV infection promoted the expression of JAG1 and DLL3. Activation of Notch-1/ DLL 3 in lungs may be associated with the differentiation of Th17 lymphocytes. CONCLUSIONS: Our data showed that activation of RSV stimulated the differentiation of Th17 in airway microenvironment through activation Notch-1/DLL3, which may be associated with the occurrence and development of RSV-induced asthma.
