Abstract：H4N2 subtype Avian influenza virus (AIV) has potential threat to the health of mammalian. To investigate the roles of inflammation-associated cytokines in pathopoiesis development of H4N2 subtype AIV, mice (Mus musculus) were inoculated intranasally with 106 fifty percent embryo infection dose (EID50) of P20 and P50 strains. At 1, 3 and 5 days after infection, the expression of anti-inflammatory cytokines (interleukin-2 (IL-2), IL-4 and IL-10), pro-inflammation cytokines (IL-1β, IL-6 and tumour necrosis factor-α (TNF-α)), monocyte chemoattractant protein-1 (MCP-1) and interferon-α (IFN-α), IFN-β, IFN-γ in lungs were detected by ELISA and qRT-PCR, respectively. The ELISA results showed that the expressions of IL-1β, IL-6, IL-10, MCP-1, IFN-γ and TNF-α were up-regulated, while the expressions of IL-2, IL-4 and IFN-α/β were down-regulated after the infection of both strains. Compared with P20 group, the expression level of IL-6 in P50 group very extremely significantly increased at 3 and 5 days post infection (P＜0.001), but the expression level of IL-10 was very extremely significantly higher than that in P20 group at 5 days post infection (P＜0.001). MCP-1 expression in P20 group was very extremely significantly lower than that in P50 group at 3 and 5 days post infection (P＜0.001). IFN-γ expression in P50 group was extremely significantly (P＜0.01) or very extremely significantly (P＜0.001) higher than that in P20 group at 1, 3 and 5 days post infection. The mRNA expression of IL-6 in P50 group showed significant increase compared to P20 group at 3 and 5 days post infection (P＜0.01). But the mRNA expression of IFN-γ and MCP-1 in both P20 and P50 groups had no significant changes post infection. To validate the role of IL-6 in pro-inflammation, mice were inoculated intravenously with anti-IL-6 neutralizing antibody after inoculated intranasally with P50 virus. The results showed that the expression levels of IL-6, MCP-1 and IFN-γ in each group were not statistically different, and inflammatory response was not alleviated, and it did not produce a protective effect on mice. This study showed that H4N2 subtype AIV was associated with inflammatory response in mice, but reduction of IL-6 could not alleviate the inflammatory response. The results provide a basis for the development of candidate drugs that can be used to prevent and control influenza A virus infection.
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