Biosciences, College of Health, Medicine and Life Sciences, Brunel University London, Uxbridge, London UB8 3PH, UK.
School of Biosciences and Technology, Vellore Institute of Technology, Vellore 632014, India.
Int J Mol Sci. 2022 Mar 11;23(6):3045. doi: 10.3390/ijms23063045.
The Influenza A virus (IAV) is a severe respiratory pathogen. C1q is the first subcomponent of the complement system's classical pathway. C1q is composed of 18 polypeptide chains. Each of these chains contains a collagen-like region located at the N terminus, and a C-terminal globular head region organized as a heterotrimeric structure (ghA, ghB and ghC). This study was aimed at investigating the complement activation-independent modulation by C1q and its individual recombinant globular heads against IAV infection. The interaction of C1q and its recombinant globular heads with IAV and its purified glycoproteins was examined using direct ELISA and far-Western blotting analysis. The effect of the complement proteins on IAV replication kinetics and immune modulation was assessed by qPCR. The IAV entry inhibitory properties of C1q and its recombinant globular heads were confirmed using cell binding and luciferase reporter assays. C1q bound IAV virions via HA, NA and M1 IAV proteins, and suppressed replication in H1N1, while promoting replication in H3N2-infected A549 cells. C1q treatment further triggered an anti-inflammatory response in H1N1 and pro-inflammatory response in H3N2-infected cells as evident from differential expression of TNF-α, NF-κB, IFN-α, IFN-β, IL-6, IL-12 and RANTES. Furthermore, C1q treatment was found to reduce luciferase reporter activity of MDCK cells transfected with H1N1 pseudotyped lentiviral particles, indicative of an entry inhibitory role of C1q against infectivity of IAV. These data appear to demonstrate the complement-independent subtype specific modulation of IAV infection by locally produced C1q.
甲型流感病毒(IAV)是一种严重的呼吸道病原体。C1q 是补体系统经典途径的第一个亚单位。C1q 由 18 条多肽链组成。每条链都含有位于 N 端的胶原样区域和组织成三聚体结构(ghA、ghB 和 ghC)的 C 端球状头部区域。本研究旨在研究 C1q 及其单独的重组球状头部对 IAV 感染的补体激活非依赖性调节。使用直接 ELISA 和远 Western 印迹分析研究了 C1q 及其重组球状头部与 IAV 及其纯化糖蛋白的相互作用。通过 qPCR 评估补体蛋白对 IAV 复制动力学和免疫调节的影响。通过细胞结合和荧光素酶报告基因测定证实了 C1q 和其重组球状头部对 IAV 进入的抑制特性。C1q 通过 IAV 的 HA、NA 和 M1 蛋白结合 IAV 病毒颗粒,并抑制 H1N1 的复制,同时促进 H3N2 感染的 A549 细胞的复制。C1q 处理进一步触发了 H1N1 中的抗炎反应和 H3N2 感染细胞中的促炎反应,这从 TNF-α、NF-κB、IFN-α、IFN-β、IL-6、IL-12 和 RANTES 的差异表达中可以看出。此外,发现 C1q 处理降低了转染有 H1N1 假型慢病毒颗粒的 MDCK 细胞的荧光素酶报告基因活性,表明 C1q 对 IAV 感染具有进入抑制作用。这些数据似乎表明局部产生的 C1q 对 IAV 感染具有补体非依赖性的亚型特异性调节作用。