Tick-borne flavivirus infection in Ixodes scapularis larvae: Development of a novel method for synchronous viral infection of ticks

Mitzel, Dana N.; Wolfinbarger, James B.; Long, Rodney; Masnick, Max; Best, Sonja M.; Bloom, Marshall E.

doi:10.1016/j.virol.2007.03.057

Cited by 44 publications

(64 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition, different concentrations of the initial TBFV inoculum may also affect TBFV replication kinetics in these organs. Mitzel et al showed that different concentrations for the LGTV inoculum affect the establishment of LGTV genome replication in I. scapularis larvae (30). Similarly, we suspect that different concentrations of the initial TBFV inoculum in female I. scapularis organ cultures could reveal varied levels of TBFV replication at different time points postinfection.…”

Section: Discussionmentioning

confidence: 84%

“…ticks (30,60,61). Infectious LGTV and POWV were produced from all three infected tick organ cultures ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Unfed, female I. scapularis ticks were obtained from a laboratory colony maintained at the Oklahoma State University Tick Rearing Facility. Ticks were maintained as described by Mitzel et al (30). Ticks were surface sterilized (79) by soaking them in 3% H 2 O 2 for 15 s followed by soaking in 70% ethanol for 15 s. They were then wicked free of excess fluid on sterile Whatman paper and subsequently dissected in sterile phosphate-buffered saline (PBS) on a sterile glass microscope slide.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Flavivirus Infection ofIxodes scapularis(Black-Legged Tick)Ex VivoOrganotypic Cultures and Applications for Disease Control

Grabowski¹,

Tsetsarkin

Long³

et al. 2017

mBio

Self Cite

View full text Add to dashboard Cite

Ixodes scapularis ticks transmit many infectious agents that cause disease, including tick-borne flaviviruses (TBFVs). TBFV infections cause thousands of human encephalitis cases worldwide annually. In the United States, human TBFV infections with Powassan virus (POWV) are increasing and have a fatality rate of 10 to 30%. Additionally, Langat virus (LGTV) is a TBFV of low neurovirulence and is used as a model TBFV. TBFV replication and dissemination within I. scapularis organs are poorly characterized, and a deeper understanding of virus biology in this vector may inform effective countermeasures to reduce TBFV transmission. Here, we describe short-term, I. scapularis organ culture models of TBFV infection. Ex vivo organs were metabolically active for 9 to 10 days and were permissive to LGTV and POWV replication. Imaging and videography demonstrated replication and spread of green fluorescent protein-expressing LGTV in the organs. Immunohistochemical staining confirmed LGTV envelope and POWV protein synthesis within the infected organs.LGTV-and POWV-infected organs produced infectious LGTV and POWV; thus, the ex vivo cultures were suitable for study of virus replication in individual organs. LGTVand POWV-infected midgut and salivary glands were subjected to double-stranded RNA (dsRNA) transfection with dsRNA to the LGTV 3= untranslated region (UTR), which reduced infectious LGTV and POWV replication, providing a proof-of-concept use of RNA interference in I. scapularis organ cultures to study the effects on TBFV replication. The results contribute important information on TBFV localization within ex vivo I. scapularis organs and provide a significant translational tool for evaluating recombinant, live vaccine candidates and potential tick transcripts and proteins for possible therapeutic use and vaccine development to reduce TBFV transmission.IMPORTANCE Tick-borne flavivirus (TBFV) infections cause neurological and/or hemorrhagic disease in humans worldwide. There are currently no licensed therapeutics or vaccines against Powassan virus (POWV), the only TBFV known to circulate in North America. Evaluating tick vector targets for antitick vaccines directed at reducing TBFV infection within the arthropod vector is a critical step in identifying efficient approaches to controlling TBFV transmission. This study characterized infection of female Ixodes scapularis tick organ cultures of midgut, salivary glands, and synganglion with the low-neurovirulence Langat virus (LGTV) and the more pathogenic POWV. Cell types of specific organs were susceptible to TBFV infection, and a difference in LGTV and POWV replication was noted in TBFV-infected organs. This tick organ culture model of TBFV infection will be useful for various applications,

show abstract

Section: Discussionmentioning

confidence: 84%

“…ticks (30,60,61). Infectious LGTV and POWV were produced from all three infected tick organ cultures ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Flavivirus Infection ofIxodes scapularis(Black-Legged Tick)Ex VivoOrganotypic Cultures and Applications for Disease Control

Grabowski¹,

Tsetsarkin

Long³

et al. 2017

mBio

Self Cite

View full text Add to dashboard Cite

show abstract

“…To investigate the ability of the viruses to infect and replicate in tick cells, infection of Vero and ISE6 cells was undertaken as previously described (Mitzel et al 2007). Both cell lines were initially infected with virus at a multiplicity of infection of 1, incubated at 348C or 378C, respectively, for 1 h, followed by addition of fresh maintenance medium.…”

Section: Virus Infection In Cell Culturementioning

confidence: 99%

“…Infection of ticks by the immersion method has been previously described (Mitzel et al 2007). Briefly, 60 larvae per group were collected in a sterile 1.5-mL screw cap centrifuge tube and pretreated by exposure to a reduced relative humidity.…”

Section: Virus Infection In Mosquitoes and Ticksmentioning

confidence: 99%

Chimeric Tick-Borne Encephalitis/Dengue Virus Is Attenuated inIxodes scapularisTicks andAedes aegyptiMosquitoes

Engel

Mitzel

Hanson

et al. 2011

Vector-Borne and Zoonotic Diseases

Self Cite

View full text Add to dashboard Cite

In an effort to derive an efficacious live attenuated vaccine against tick-borne encephalitis, we generated a chimeric virus bearing the structural protein genes of a Far Eastern subtype of tick-borne encephalitis virus (TBEV) on the genetic background of recombinant dengue 4 (DEN4) virus. Introduction of attenuating mutations into the TBEV envelope protein gene, as well as the DEN4 NS5 protein gene and 3 0 noncoding region in the chimeric genome, results in decreased neurovirulence and neuroinvasiveness in mice, and restricted replication in mouse brain. Since TBEV and DEN4 viruses are transmitted in nature by ticks and mosquitoes, respectively, it was of interest to investigate the infectivity of the chimeric virus for both arthropod vectors. Therefore, parental and chimeric viruses were tested for growth in mosquito and tick cells and for oral infection in vivo. Although all chimeric viruses demonstrated moderate levels of replication in C6/36 mosquito cells, they were unable to replicate in ISE6 tick cells. Further, the chimeric viruses were unable to infect or replicate in Aedes aegypti mosquitoes and Ixodes scapularis tick larvae. The poor infectivity for both potential vectors reinforces the safety of chimeric virus-based vaccine candidates for the environment and for use in humans.

show abstract

Evidence of lumpy skin disease virus over-wintering by transstadial persistence in Amblyomma hebraeum and transovarial persistence in Rhipicephalus decoloratus ticks

et al. 2013

View full text Add to dashboard Cite

Lumpy skin disease is a debilitating cattle disease caused by the lumpy skin disease virus (LSDV), belonging to the genus Capripoxvirus. Epidemics of the disease usually occur in summer, when insect activity is high. Limited information is available on how LSDV persists during inter-epidemic periods. Transmission of LSDV by mosquitoes such as Aedes aegypti has been shown to be mechanical, there is no carrier state in cattle and the role of wildlife in the epidemiology of the disease seems to be of minor importance. Recent studies in ticks have shown transstadial persistence of LSDV in Rhipicephalus appendiculatus and Amblyomma hebraeum as well as transovarial persistence of the virus in Rhipicephalus decoloratus, R. appendiculatus and A. hebraeum. The over-wintering of ticks off the host as part of their life cycles is well known: A. hebraeum and R. appendiculatus over-winter, for example, on the ground as engorged nymphs/unfed (emergent) adults while R. decoloratus over-winters on the ground as engorged females. In this study, transstadial and transovarial persistence of LSDV from experimentally infected A. hebraeum nymphs and R. decoloratus females after exposure to cold temperatures of 5 °C at night and 20 °C during the day for 2 months was reported. This observation suggests possible over-wintering of the virus in these tick species.

show abstract

Tick-borne flavivirus infection in Ixodes scapularis larvae: Development of a novel method for synchronous viral infection of ticks

Cited by 44 publications

References 24 publications

Flavivirus Infection ofIxodes scapularis(Black-Legged Tick)Ex VivoOrganotypic Cultures and Applications for Disease Control

Flavivirus Infection ofIxodes scapularis(Black-Legged Tick)Ex VivoOrganotypic Cultures and Applications for Disease Control

Chimeric Tick-Borne Encephalitis/Dengue Virus Is Attenuated inIxodes scapularisTicks andAedes aegyptiMosquitoes

Evidence of lumpy skin disease virus over-wintering by transstadial persistence in Amblyomma hebraeum and transovarial persistence in Rhipicephalus decoloratus ticks

Contact Info

Product

Resources

About