Moreira Étori Aguiar, Locher Samira, Kolesnikova Larissa, Bolte Hardin, Aydillo Teresa, García-Sastre Adolfo, Schwemmle Martin, Zimmer Gert
Institute of Virology, University Medical Center Freiburg, D-79104 Freiburg, Germany.
Spemann Graduate School of Biology and Medicine, University of Freiburg, D-79104 Freiburg, Germany.
Proc Natl Acad Sci U S A. 2016 Nov 8;113(45):12797-12802. doi: 10.1073/pnas.1608821113. Epub 2016 Oct 24.
Two novel influenza A-like viral genome sequences have recently been identified in Central and South American fruit bats and provisionally designated "HL17NL10" and "HL18NL11." All efforts to isolate infectious virus from bats or to generate these viruses by reverse genetics have failed to date. Recombinant vesicular stomatitis virus (VSV) encoding the hemagglutinin-like envelope glycoproteins HL17 or HL18 in place of the VSV glycoprotein were generated to identify cell lines that are susceptible to bat influenza A-like virus entry. More than 30 cell lines derived from various species were screened but only a few cell lines were found to be susceptible, including Madin-Darby canine kidney type II (MDCK II) cells. The identification of cell lines susceptible to VSV chimeras allowed us to recover recombinant HL17NL10 and HL18NL11 viruses from synthetic DNA. Both influenza A-like viruses established a productive infection in MDCK II cells; however, HL18NL11 replicated more efficiently than HL17NL10 in this cell line. Unlike conventional influenza A viruses, bat influenza A-like viruses started the infection preferentially at the basolateral membrane of polarized MDCK II cells; however, similar to conventional influenza A viruses, bat influenza A-like viruses were released primarily from the apical site. The ability of HL18NL11 or HL17NL10 viruses to infect canine and human cells might reflect a zoonotic potential of these recently identified bat viruses.
最近在中美洲和南美洲的果蝠中发现了两种新型甲型流感样病毒基因组序列,暂命名为“HL17NL10”和“HL18NL11”。迄今为止,所有从蝙蝠中分离出感染性病毒或通过反向遗传学产生这些病毒的努力均告失败。构建了编码血凝素样包膜糖蛋白HL17或HL18以取代水泡性口炎病毒(VSV)糖蛋白的重组VSV,以鉴定对蝙蝠甲型流感样病毒进入敏感的细胞系。对来自不同物种的30多种细胞系进行了筛选,但仅发现少数细胞系敏感,包括II型犬肾传代细胞(MDCK II)。对VSV嵌合体敏感的细胞系的鉴定使我们能够从合成DNA中回收重组HL17NL10和HL18NL11病毒。两种甲型流感样病毒均在MDCK II细胞中建立了有效感染;然而,在该细胞系中,HL18NL11的复制效率高于HL17NL10。与传统甲型流感病毒不同,蝙蝠甲型流感样病毒优先在极化的MDCK II细胞的基底外侧膜开始感染;然而,与传统甲型流感病毒类似,蝙蝠甲型流感样病毒主要从顶端部位释放。HL18NL11或HL17NL10病毒感染犬类和人类细胞的能力可能反映了这些最近发现的蝙蝠病毒的人畜共患病潜力。