Infection of Caenorhabditis elegans with Vesicular Stomatitis Virus via Microinjection

Martín, A.; Rex, Emily A.; Ishidate, Takao; Lin, Rueyling; Gammon, Don B.

doi:10.21769/bioprotoc.2617

Cited by 5 publications

(4 citation statements)

References 14 publications

(31 reference statements)

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“…There are several natural enteric viruses that have been recently isolated from wild-caught nematodes, including Orsay virus, facilitating the study of host-virus interactions [185]. Further, C. elegans are potentially amenable to infection with non-natural nematode viruses via microinjection [186]. C. elegans is thus a potential model to identify host-pathogen interactions during enteric virus co-infections.…”

Section: Caenorhabditis Elegansmentioning

confidence: 99%

Enteric Viral Co-Infections: Pathogenesis and Perspective

Makimaa

Ingle

Baldridge

2020

Viruses

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Enteric viral co-infections, infections involving more than one virus, have been reported for a diverse group of etiological agents, including rotavirus, norovirus, astrovirus, adenovirus, and enteroviruses. These pathogens are causative agents for acute gastroenteritis and diarrheal disease in immunocompetent and immunocompromised individuals of all ages globally. Despite virus–virus co-infection events in the intestine being increasingly detected, little is known about their impact on disease outcomes or human health. Here, we review what is currently known about the clinical prevalence of virus–virus co-infections and how co-infections may influence vaccine responses. While experimental investigations into enteric virus co-infections have been limited, we highlight in vivo and in vitro models with exciting potential to investigate viral co-infections. Many features of virus–virus co-infection mechanisms in the intestine remain unclear, and further research will be critical.

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Section: Caenorhabditis Elegansmentioning

confidence: 99%

Enteric Viral Co-Infections: Pathogenesis and Perspective

Makimaa

Ingle

Baldridge

2020

Viruses

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“…Worms were injected with VSV∆G-AFF-1 that was pre-incubated with αVSV-G antibody (see materials and methods and Figure S4), VSV∆G-G (as a positive control) or DMEM medium (as negative control). We used drh-1 (-) (Dicer Related Helicase -1) mutant worms, as they are more susceptible to VSV infection and do not have any observable phenotypes [7,34]. Taken that VSV-G works unilaterally, while AFF-1 and EFF-1 fusogens have to be present in both fusing membranes [6,16,18,30], we hypothesized that VSV∆G-G and VSV∆G-AFF-1 will produce different infection patterns.…”

Section: Vsv∆g-aff-1 and Vsv∆g-g Infect Different Cells In C Elegansmentioning

confidence: 99%

“…Scoring viral infection using fluorescence assay was performed as described [7,34] with some modifications. Briefly, 48-72 hours post injection worms were processed for live imaging as described above.…”

Section: Scoring Viral Infectionmentioning

confidence: 99%

“…To date, however, VSVΔG coated with AFF-1, EFF-1 or any other non-viral fusogen have not been tested for infection in a living organism so far. Recently VSVΔG-G was demonstrated to infect living C. elegans [7,34]. Given that: (i) VSVΔG-AFF-1 requires a fusogen on the target cell for infection, (ii) a detailed cellular atlas of AFF-1 and EFF-1 expression and function in C. elegans is available and (iii) C. elegans eff-1 and aff-1 mutants are available, we test whether VSVΔG-AFF-1 can be exploited as a cell-specific delivery system in C. elegans.…”

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confidence: 99%

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EFF-1 promotes muscle fusion, paralysis and retargets infection by AFF-1-coated viruses inC. elegans

Meledin

Matveev

et al. 2020

Preprint

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AFF-1-coated virus infects skin cells and rarely muscle cells in C. elegans• Epithelial cell-specific infection requires eff-1 activity • Pseudoviral infection depends on the amount of eff-1 in target cells • Ectopic EFF-1 in muscle cells induces their fusion and viral infectionAbstract Viral-based vectors are widely used for fundamental and translational research; however, production of a tissue-specific targeting vector is often a challenging step. Here, we develop a cell type-specific delivery system, based on vesicular stomatitis virus lacking the viral fusion protein (fusogen) and pseudotyped with AFF-1, a C. elegans cell-to-cell bilateral fusogen from the fusexin family. Following injection of AFF-1-coated virus into the C. elegans pseudocoelom that functions as a circulatory system, we observe specific infection of different cells that express EFF-1 or AFF-1 fusogens. EFF-1-expressing epidermal cells are preferentially infected, whereas mononucleated body wall muscles (BWMs) that do not express any fusogen are rarely infected. The epidermal cells infection is abolished when the host cells do not express EFF-1, while ectopic expression of EFF-1 on BWMs induces their abnormal cell-cell fusion, produces paralyzed-dumpy animals and enhances muscle infection by AFF-1-coated viruses. Thus, our data uncover that AFF-1-coated pseudotyped viruses can be retargeted to desired cells in a controlled manner and serve as a cell-specific delivery system in nematodes with potential applications in regenerative processes, therapies such as dendritic reconnection following injuries, and studies of diverse cellular and viral fusogens.VSV-G is a class III viral fusogen whereas the Caenorhabditis elegans EFF-1 and AFF-1, which fuse somatic cells during development, are structural homologs of class II viral proteins and gamete HAP2(GCS1) fusogens from the fusexin family [14][15][16][17][18][19][20][21][22]. EFF-1 and AFF-1 also participate in maintaining neuronal architecture and neuronal reconnection following injury [23][24][25][26][27]. Studied viral glycoproteins, including VSV-G, use a unilateral fusion mechanism that depends on the expression of receptors only on the target cells [28,29]. In contrast, EFF-1 and AFF-1 are bilateral fusogens-their presence is required on the membranes of both apposing cells to mediate fusion [6,16,18,30,31]. These two fusogens can act in either a homotypic or a heterotypic manner [6,16,18,21] and mediate heterotypic cell fusion of Sf9 insect and Baby Hamster Kidney (BHK) cells [6,16]. Finally, VSV viruses containing a GFP substituting the VSV-G coding sequence (VSV∆G) [32,33] that are coated with AFF-1 (VSV∆G-AFF-1) specifically infect AFF-1 or EFF-1 expressing BHK cells [6]. Thus, in contrast to the pseudotyped virus coated with the native, unilateral G glycoprotein (VSVΔG-G), infection by VSVΔG-AFF-1 requires fusogen expression on the host cell membrane. To date, however, VSVΔG coated with AFF-1, EFF-1 or any other non-viral fusogen have not been tested for infection in a living organism ...

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Nematodes

Bodri

2021

Invertebrate Medicine

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Infection of Caenorhabditis elegans with Vesicular Stomatitis Virus via Microinjection

Cited by 5 publications

References 14 publications

Enteric Viral Co-Infections: Pathogenesis and Perspective

Enteric Viral Co-Infections: Pathogenesis and Perspective

EFF-1 promotes muscle fusion, paralysis and retargets infection by AFF-1-coated viruses inC. elegans

Nematodes

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