牛副流感病毒3(BPIV3)是幼牛和成年牛呼吸道疾病的关键病原体。没有针对BPIV3感染的特定疗法。了解病毒的内化途径将为抗病毒治疗的发展提供新的策略。这里,使用RNA沉默和药物抑制剂分析了BPIV3进入HeLa细胞的机制.用高渗培养基处理HeLa细胞可防止BPIV3内化。这些结果表明BPIV3通过受体介导的内吞作用进入HeLa细胞。此外,通过MβCD处理去除细胞膜胆固醇阻碍了病毒渗透,但不妨碍病毒复制.此外,BPIV3感染可通过用dynamore或氯丙嗪(CPZ)预处理或敲除动力学II或网格蛋白重链来抑制。然而,病毒进入不受制霉菌素的影响,EIPA,Wortmannin,或细胞松弛素D治疗或caveolin-1敲低。这些数据表明,BPIV3进入HeLa细胞取决于网格蛋白介导的内吞作用,而不取决于caveolae介导的内吞作用或巨生胞吞作用途径。许多病毒被运送到内体,它们提供了酸性环境,并在与原代内吞囊泡分离后释放它们的基因组。然而,我们发现BPIV3感染需要内体组织蛋白酶,但不是低pH值。总之,我们展示,第一次,BPIV3通过网格蛋白介导的内吞途径进入HeLa细胞,对副粘病毒科的侵袭机制提出了新的见解。
Bovine parainfluenza virus 3 (
BPIV3) is a crucial causative agent of respiratory disease in young and adult cattle. No specific therapies are available for
BPIV3 infection. Understanding the internalization pathway of the virus will provide a new strategy for the development of antiviral therapy. Here, the mechanism of BPIV3 entry into HeLa cells was analyzed using RNA silencing and pharmacological inhibitors. Treatment of HeLa cells with hypertonic medium prevented
BPIV3 internalization. These results indicated that
BPIV3 entered HeLa cells via receptor-mediated endocytosis. Moreover, removing cell membrane cholesterol through MβCD treatment hampered viral penetration but not viral replication. In addition, BPIV3 infection was inhibited by pretreatment with dynasore or chlorpromazine (CPZ) or knockdown of dynamin II or clathrin heavy chain. However, virus entry was unaffected by nystatin, EIPA, wortmannin, or cytochalasin D treatment or caveolin-1 knockdown. These data demonstrated that the entry of BPIV3 into HeLa cells was dependent on clathrin-mediated endocytosis but not on caveolae-mediated endocytosis or the macropinocytosis pathway. Many viruses are transported to endosomes, which provide an acidic environment and release their genome upon separation from primary endocytic vesicles. However, we found that
BPIV3 infection required endosomal cathepsins, but not a low pH. In summary, we show, for the first time, that BPIV3 enters HeLa cells through the clathrin-mediated endocytosis pathway, presenting novel insights into the invasion mechanism of Paramyxoviridae.