Human influenza virus infection and apoptosis induction in human vascular endothelial cells
Human influenza virus infection and apoptosis induction in human vascular endothelial cells
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DOI：
10.1002/jmv.21185
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发表时间：
2008-06-01
影响因子：
12.7
通讯作者：
Hosoya, Mitsuaki
中科院分区：
文献类型：
--
作者：
Sumikoshi, Makoto;Hashimoto, Koichi;Hosoya, Mitsuaki
流感病毒感染引起的急性脑病主要通过血管源性水肿导致脑部和全身器官衰竭，并伴有高水平的炎症细胞因子，如血液肿瘤坏死因子（TNF）-α和白细胞介素（IL）-6以及细胞色素c凋亡标记物。一种高毒力的禽流感病毒株通过感染全身器官的血管内皮细胞并诱导其中的细胞凋亡而导致鸡致命。为了验证人流感病毒在感染的人血管内皮细胞中诱导凋亡的可能性，使用组织培养方法检查纯化的流感病毒感染的人脐静脉内皮细胞（HUVEC）。当用 TNF-α 预处理时，流感病毒（菲律宾株，H3N2）促进 TNF-α 诱导的 HUVEC 凋亡。在没有 TNF-α 的情况下，感染菲律宾毒株的 HUVEC 中的病毒复制得到证实。通过使用组织培养感染剂量（TCID）方法、免疫组织化学和实时PCR测量培养物上清液中感染性病毒的量。感染后24小时，受感染的HUVEC中流感病毒基因组的数量比病毒吸附后增加了约五倍。使用TUNEL方法观察到许多TUNEL阳性流感病毒感染的HUVEC。此外，通过免疫荧光染色，在流感病毒感染的细胞中也检测到了裂解的 caspase 3。这些结果表明，人流感病毒可以感染人血管内皮细胞并在其中复制并诱导其中的细胞凋亡。
Acute encephalopathy accompanying influenza virus infection results in brain and systemic organ failure mainly through vasogenic edema with high levels of inflammatory cytokines, such as blood tumor necrosis factor (TNF)-alpha and interleukin (IL)-6, as well as the cytochrome c apoptosis marker. A highly virulent strain of avian influenza virus causes fatal infection in chickens by infecting vascular endothelial cells in systemic organs, inducing apoptosis therein. To verify the possibility of apoptosis induction by human influenza virus in infected human vascular endothelial cells, purified influenza virus-infected human umbilical vein endothelial cells (HUVECs) were examined using a tissue culture method. When pre-treated with TNF-alpha, influenza virus (Philippine strain, H3N2) promoted TNF-a induced apoptosis of HUVECs. Viral replication was confirmed in HUVECs infected with the Philippine strain in the absence of TNF-alpha. by measurement of the amount of infective virus in the culture supernatant using the tissue culture infectious dose (TCID) method, immunohistochemistry and real-time PCR. The number of influenza virus genomes in the infected HUVECs at 24 hr post-infection increased about fivefold compared to that just after virus adsorption. Many TUNEL-positive influenza virus-infected HUVECs were observed using the TUNEL method. Furthermore, cleaved caspase 3 was also detected in influenza virus-infected cells by immunofluorescence staining. These results demonstrated that human influenza virus can infect and replicate in human vascular endothelial cells and induce apoptosis therein.