Role of IL10 signaling in the circadian control of host response to Influenza infection. Role of IL10 signaling in the circadian control of host response to Influenza infection

The circadian clock confers a time-of-day specific protection against influenza A Virus (IAV) infection by regulating the host response and limiting immunopathology. IL10 is a cytokine with significant immunoregulatory functions; however, the role of IL10 in IAV is unclear, with reports of both benefit and harm from its blockade early during the infection. Since the expression of IL10R shows diurnal rhythmicity in the lungs, we tested if IL10 signaling contributes to time-of-day specific protection against IAV. Overall design: Mice were intranasally infected with 30-35 PFU of mouse-adapted H1N1 (PR8) at either ZT11 or ZT23. To block IL-10 signaling, mice were treated with either IL-10R1a monoclonal antibody (Clone 1B1.3A, BioXCell BE0050) or polyclonal IgG (Clone MOPC-21, BioXCell BE0083) during the infection. Mice were treated with 100ug/10uL of IL10R1 MAb administered intranasally D0 in 10uL PBS, 150ug/100uL of IL10R1 Mab administered intraperitoneally on day 3 in 100uL of PBS, and 100ug/100uL of IL10R1 MAb administered i.p. D5.

昼夜节律时钟可通过调控宿主免疫应答并限制免疫病理损伤，为甲型流感病毒（Influenza A Virus, IAV）感染提供日间特异性的保护作用。白细胞介素10（Interleukin 10, IL10）是一类具有重要免疫调节功能的细胞因子，但其在甲型流感病毒感染中的作用尚不明确，现有研究对感染早期阻断IL10后的效果存在利弊不一的报道。鉴于IL10受体（IL10R）的表达在肺部呈现昼夜节律性，本研究验证了IL10信号通路是否参与介导针对甲型流感病毒感染的日间特异性保护作用。

整体实验设计：分别于昼夜节律时间点ZT11与ZT23时段，以30-35个噬斑形成单位（Plaque Forming Unit, PFU）的小鼠适应性H1N1（PR8毒株）经鼻感染小鼠。为阻断IL-10信号通路，感染期间分别向小鼠给予相应抗体处理：经鼻滴注100μg/10μL IL10R1a单克隆抗体（克隆号1B1.3A，购自BioXCell，货号BE0050）或同型对照多克隆IgG（克隆号MOPC-21，购自BioXCell，货号BE0083）。具体给药方案为：第0天经鼻给予10μL磷酸盐缓冲液（Phosphate Buffered Saline, PBS）配制的100μg IL10R1单克隆抗体，第3天经腹腔给予100μL磷酸盐缓冲液配制的150μg IL10R1单克隆抗体，第5天经腹腔给予100μL磷酸盐缓冲液配制的100μg IL10R1单克隆抗体。