Entry of the Varicellovirus Canid herpesvirus 1 into Madin–Darby canine kidney epithelial cells is pH-independent and occurs via a macropinocytosis-like mechanism but without increase in fluid uptake

IF 2.6 2区 生物学 Q3 CELL BIOLOGY
Mohamed Eisa, Hamza Loucif, Julien van Grevenynghe, Angela Pearson
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引用次数: 4

Abstract

Canid herpesvirus 1 (CHV-1) is a Varicellovirus that causes self-limiting infections in adult dogs but morbidity and mortality in puppies. Using a multipronged approach, we discovered the CHV-1 entry pathway into Madin–Darby canine kidney (MDCK) epithelial cells. We found that CHV-1 triggered extensive host cell membrane lamellipodial ruffling and rapid internalisation of virions in large, uncoated vacuoles, suggestive of macropinocytosis. Treatment with inhibitors targeting key macropinocytosis factors, including inhibitors of Na+/H+ exchangers, F-actin, myosin light-chain kinase, protein kinase C, p21-activated kinase, phosphatidylinositol-3-kinase and focal adhesion kinase, significantly reduced viral replication. Moreover, the effect was restricted to exposure to the inhibitors early in infection, confirming a role for the macropinocytic machinery during entry. The profile of inhibitors also suggested a role for signalling via integrins and receptor tyrosine kinases in viral entry. In contrast, inhibitors of clathrin, caveolin, microtubules and endosomal acidification did not affect CHV-1 entry into MDCK cells. We found that the virus colocalised with the fluid-phase uptake marker dextran; however, surprisingly, CHV-1 infection did not enhance the uptake of dextran. Thus, our results indicate that CHV-1 uses a macropinocytosis-like, pH-independent entry pathway into MDCK cells, which nevertheless is not based on stimulation of fluid uptake.

Take Aways

  • CHV-1 enters epithelial cells via a macropinocytosis-like mechanism.
  • CHV-1 induces extensive lamellipodial ruffling.
  • CHV-1 entry into MDCK cells is pH-independent.

Abstract Image

犬疱疹病毒1型水痘病毒进入Madin-Darby犬肾上皮细胞是不依赖于ph值的,并通过类似巨噬细胞增多的机制发生,但不增加液体摄取
犬疱疹病毒1型(CHV-1)是一种水痘病毒,在成年犬中引起自限性感染,但在幼犬中发病和死亡。通过多管齐下的方法,我们发现了CHV-1进入Madin-Darby犬肾(MDCK)上皮细胞的途径。我们发现CHV-1引发宿主细胞膜广泛的板状褶皱和病毒粒子在大的、未被包裹的液泡中的快速内化,提示巨噬细胞增多症。以Na+/H+交换物、f -肌动蛋白、肌球蛋白轻链激酶、蛋白激酶C、p21活化激酶、磷脂酰肌醇-3激酶和黏附斑激酶为靶点的抑制剂治疗,可显著降低病毒复制。此外,这种作用仅限于感染早期暴露于抑制剂,证实了进入时巨噬细胞机制的作用。抑制剂的特征也表明在病毒进入过程中通过整合素和受体酪氨酸激酶信号传导的作用。相比之下,网格蛋白、小窝蛋白、微管和内体酸化抑制剂对CHV-1进入MDCK细胞没有影响。我们发现病毒与液相摄取标记葡聚糖共定位;然而,令人惊讶的是,CHV-1感染并没有增强右旋糖酐的摄取。因此,我们的研究结果表明,CHV-1使用类似巨噬细胞症的、不依赖ph的进入MDCK细胞的途径,然而,这不是基于刺激液体摄取。CHV-1通过巨噬细胞样机制进入上皮细胞。CHV-1诱导广泛的板足褶。CHV-1进入MDCK细胞与ph无关。
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来源期刊
Cellular Microbiology
Cellular Microbiology 生物-微生物学
CiteScore
9.70
自引率
0.00%
发文量
26
审稿时长
3 months
期刊介绍: Cellular Microbiology aims to publish outstanding contributions to the understanding of interactions between microbes, prokaryotes and eukaryotes, and their host in the context of pathogenic or mutualistic relationships, including co-infections and microbiota. We welcome studies on single cells, animals and plants, and encourage the use of model hosts and organoid cultures. Submission on cell and molecular biological aspects of microbes, such as their intracellular organization or the establishment and maintenance of their architecture in relation to virulence and pathogenicity are also encouraged. Contributions must provide mechanistic insights supported by quantitative data obtained through imaging, cellular, biochemical, structural or genetic approaches.
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