Kolokoltsova Olga A, Grant Ashley M, Huang Cheng, Smith Jennifer K, Poussard Allison L, Tian Bing, Brasier Allan R, Peters Clarence J, Tseng Chien-Te Kent, de la Torre Juan C, Paessler Slobodan
Department of Pathology, University of Texas Medical Branch (UTMB), Galveston, Texas, United States of America.
Internal Med-Endocrinology, UTMB, Galveston, Texas, United States of America.
PLoS One. 2014 Jun 11;9(6):e99610. doi: 10.1371/journal.pone.0099610. eCollection 2014.
Junin virus (JUNV) is the etiological agent of Argentine hemorrhagic fever (AHF), a human disease with a high case-fatality rate. It is widely accepted that arenaviral infections, including JUNV infections, are generally non-cytopathic. In contrast, here we demonstrated apoptosis induction in human lung epithelial carcinoma (A549), human hepatocarcinoma and Vero cells upon infection with the attenuated Candid#1 strain of, JUNV as determined by phosphatidylserine (PS) translocation, Caspase 3 (CASP3) activation, Poly (ADP-ribose) polymerase (PARP) cleavage and/or chromosomal DNA fragmentation. Moreover, as determined by DNA fragmentation, we found that the pathogenic Romero strain of JUNV was less cytopathic than Candid#1 in human hepatocarcinoma and Vero, but more apoptotic in A549 and Vero E6 cells. Additionally, we found that JUNV-induced apoptosis was enhanced by RIG-I signaling. Consistent with the previously reported role of RIG-I like helicase (RLH) signaling in initiating programmed cell death, we showed that cell death or DNA fragmentation of Candid#1-infected A549 cells was decreased upon siRNA or shRNA silencing of components of RIG-I pathway in spite of increased virus production. Similarly, we observed decreased DNA fragmentation in JUNV-infected human hepatocarcinoma cells deficient for RIG-I when compared with that of RIG-I-competent cells. In addition, DNA fragmentation detected upon Candid#1 infection of type I interferon (IFN)-deficient Vero cells suggested a type I IFN-independent mechanism of apoptosis induction in response to JUNV. Our work demonstrated for the first time apoptosis induction in various cells of mammalian origin in response to JUNV infection and partial mechanism of this cell death.
胡宁病毒(JUNV)是阿根廷出血热(AHF)的病原体,这是一种病死率很高的人类疾病。人们普遍认为包括JUNV感染在内的沙粒病毒感染通常无细胞病变效应。相比之下,我们在此证明,用JUNV减毒Candid#1株感染人肺上皮癌(A549)细胞、人肝癌细胞和Vero细胞后,会诱导细胞凋亡,这是通过磷脂酰丝氨酸(PS)易位、半胱天冬酶3(CASP3)激活、聚(ADP-核糖)聚合酶(PARP)裂解和/或染色体DNA片段化来确定的。此外,通过DNA片段化检测发现,JUNV的致病性罗梅罗毒株在人肝癌细胞和Vero细胞中的细胞病变效应比Candid#1株弱,但在A549细胞和Vero E6细胞中诱导凋亡的能力更强。此外,我们发现RIG-I信号增强了JUNV诱导的细胞凋亡。与之前报道的RIG-I样解旋酶(RLH)信号在启动程序性细胞死亡中的作用一致,我们发现,尽管病毒产量增加,但在对RIG-I途径成分进行siRNA或shRNA沉默后,Candid#1感染的A549细胞的细胞死亡或DNA片段化减少。同样,与具有RIG-I功能的细胞相比,我们观察到JUNV感染的缺乏RIG-I的人肝癌细胞中的DNA片段化减少。此外,在I型干扰素(IFN)缺陷的Vero细胞被Candid#1感染后检测到的DNA片段化表明,存在一种不依赖I型干扰素的JUNV诱导凋亡机制。我们的研究首次证明了JUNV感染可诱导多种哺乳动物来源的细胞发生凋亡,并揭示了这种细胞死亡的部分机制。