tetano
Editor, Senior Moderator
Virol J
. 2020 Jul 6;17(1):92.
doi: 10.1186/s12985-020-01341-x.
Pulmonary Endothelium-Derived PD-L1 Induced by the H9N2 Avian Influenza Virus Inhibits the Immune Response of T Cells
Qian Zhang[SUP] 1 [/SUP], Xiang Mu[SUP] 2 [/SUP], Hong Dong[SUP] 2 [/SUP], Ge Hu[SUP] 2 [/SUP], Tao Zhang[SUP] 2 [/SUP], Cheng He[SUP] 3 [/SUP], Naila Siddique[SUP] 4 [/SUP]
Affiliations
Abstract
Background: The PD-1/PD-L1 pathway is an inhibitory signaling pathway that maintains the balance between the immune response and immunotolerance, and its overactivation in cancer and viral infections inhibits T cell function. The target cells of various viruses, microvascular endothelial cells (MECs) have been shown to be key regulatory points in immune regulation and virion diffusion in vivo during infection with multiple influenza virus subtypes. Furthermore, avian influenza virus (AIV) infection can induce immunosuppression by causing imbalances in immune responses and immune organ damage. Thus, the aim of this study was to investigate whether the H9N2 virus inhibited the immune function of T cells that migrated across MECs by upregulating PD-L1 expression on MECs.
Methods: The susceptibility of rat pulmonary microvascular endothelial cells (RPMECs) to the H9N2 virus was evaluated by a plaque-forming assay and immunofluorescence staining. Then, we quantified the mRNA and protein levels of PD-L1 in RPMECs induced by H9N2 virus infection using quantitative real-time PCR and flow cytometry. The interaction between the activated T cells and RPMECs infected with the H9N2 virus was revealed using a coculture system. The effect of endothelial-derived PD-L1 on T cell function was investigated by using ELISA and flow cytometry with or without a PD-L1-specific antibody.
Results: Surface staining and the plaque-forming assay showed that the H9N2 virus infected and replicated in RPMECs. Both the PD-L1 mRNA level and PD-L1 protein level were upregulated in RPMECs infected with the H9N2 virus. H9N2 virus-induced PD-L1 expression significantly reduced the secretions of IL-2, IFN-γ and granzyme B and perforin expression in T cells. The above data were significantly increased after treatment with an anti-PD-L1 antibody, confirming the above mentioned findings. In addition, the induction of PD-L1 expression decreased the proliferative capacity of the cocultured T cells but did not affect the apoptosis rate of T cells.
Conclusions: Taken together, the results suggest that the H9N2 virus is able to inhibit the T cell immune response by upregulating PD-L1 expression in pulmonary microvascular endothelial cells.
Keywords: H9N2 virus; PD-L1; Primary microvascular endothelial cells; T cell immune response.
. 2020 Jul 6;17(1):92.
doi: 10.1186/s12985-020-01341-x.
Pulmonary Endothelium-Derived PD-L1 Induced by the H9N2 Avian Influenza Virus Inhibits the Immune Response of T Cells
Qian Zhang[SUP] 1 [/SUP], Xiang Mu[SUP] 2 [/SUP], Hong Dong[SUP] 2 [/SUP], Ge Hu[SUP] 2 [/SUP], Tao Zhang[SUP] 2 [/SUP], Cheng He[SUP] 3 [/SUP], Naila Siddique[SUP] 4 [/SUP]
Affiliations
- PMID: 32631356
- DOI: 10.1186/s12985-020-01341-x
Abstract
Background: The PD-1/PD-L1 pathway is an inhibitory signaling pathway that maintains the balance between the immune response and immunotolerance, and its overactivation in cancer and viral infections inhibits T cell function. The target cells of various viruses, microvascular endothelial cells (MECs) have been shown to be key regulatory points in immune regulation and virion diffusion in vivo during infection with multiple influenza virus subtypes. Furthermore, avian influenza virus (AIV) infection can induce immunosuppression by causing imbalances in immune responses and immune organ damage. Thus, the aim of this study was to investigate whether the H9N2 virus inhibited the immune function of T cells that migrated across MECs by upregulating PD-L1 expression on MECs.
Methods: The susceptibility of rat pulmonary microvascular endothelial cells (RPMECs) to the H9N2 virus was evaluated by a plaque-forming assay and immunofluorescence staining. Then, we quantified the mRNA and protein levels of PD-L1 in RPMECs induced by H9N2 virus infection using quantitative real-time PCR and flow cytometry. The interaction between the activated T cells and RPMECs infected with the H9N2 virus was revealed using a coculture system. The effect of endothelial-derived PD-L1 on T cell function was investigated by using ELISA and flow cytometry with or without a PD-L1-specific antibody.
Results: Surface staining and the plaque-forming assay showed that the H9N2 virus infected and replicated in RPMECs. Both the PD-L1 mRNA level and PD-L1 protein level were upregulated in RPMECs infected with the H9N2 virus. H9N2 virus-induced PD-L1 expression significantly reduced the secretions of IL-2, IFN-γ and granzyme B and perforin expression in T cells. The above data were significantly increased after treatment with an anti-PD-L1 antibody, confirming the above mentioned findings. In addition, the induction of PD-L1 expression decreased the proliferative capacity of the cocultured T cells but did not affect the apoptosis rate of T cells.
Conclusions: Taken together, the results suggest that the H9N2 virus is able to inhibit the T cell immune response by upregulating PD-L1 expression in pulmonary microvascular endothelial cells.
Keywords: H9N2 virus; PD-L1; Primary microvascular endothelial cells; T cell immune response.