Antivirus activity, but not thiolreductase activity, is conserved in interferon-gamma-inducible GILT protein in arthropod.

We have previously reported that gamma-interferon inducible lysosomal thiolreductase (GILT) functions as a host defense factor against retroviruses by digesting disulfide bonds on viral envelope proteins. GILT is widely conserved even in plants and fungi as well as animals. The thiolreductase active site of mammalian GILT is composed of a CXXC amino acid motif, whereas the C-terminal cysteine residue is changed to serine in arthropods including shrimps, crabs, and flies. GILT from Penaeus monodon (PmGILT) also has the CXXS motif instead of the CXXC active site. We demonstrate here that a human GILT mutant (GILT C75S) with the CXXS motif and PmGILT significantly inhibit amphotropic murine leukemia virus vector infection in human cells without alterning its expression level and lysosomal localization, showing that the C-terminal cysteine residue of the active site is not required for the antiviral activity. We have reported that human GILT suppresses HIV-1 particle production by digestion of disulfide bonds on CD63. However, GILT C75S mutant and PmGILT did not digest CD63 disulfide bonds, and had no effect on HIV-1 virion production, suggesting that they do not have thiolreductase activity. Taken together, this study found that antiviral activity, but not thiolreductase activity, is conserved in arthropod GILT proteins. This finding provides a new insight that the common function of GILT is antiviral activity in many animals.

Revisiting an old friend: new findings in alphavirus structure and assembly.

Alphaviruses are transmitted by an arthropod vector to a vertebrate host. The disease pathologies, cellular environments, immune responses, and host factors are very different in these organisms. Yet, the virus is able to infect, replicate, and assemble into new particles in these two animals using one set of genetic instructions. The balance between conserved mechanisms and unique strategies during virus assembly is critical for fitness of the virus. In this review, we discuss new findings in receptor binding, polyprotein topology, nucleocapsid core formation, and particle budding that have emerged in the last five years and share opinions on how these new findings might answer some questions regarding alphavirus structure and assembly.

ICTV Virus Taxonomy Profile: Solinviviridae.

Solinviviridae is a family of picorna/calici-like viruses with non-segmented, linear, positive-sense RNA genomes of approximately 10-11 kb. Unusually, their capsid proteins are encoded towards the 3'-end of the genome where they can be expressed both from a subgenomic RNA and as an extension of the replication (picorna-like helicase-protease-polymerase) polyprotein. Members of two species within the family infect ants, but related unclassified virus sequences derive from a large variety of insects and other arthropods. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the Solinviviridae, which is available at www.ictv.global/report/solinviviridae.

Assessment of the ability of V920 recombinant vesicular stomatitis-Zaire ebolavirus vaccine to replicate in relevant arthropod cell cultures and vector species.

V920, rVSVΔG-ZEBOV-GP, is a recombinant vesicular stomatitis-Zaire ebolavirus vaccine which has shown an acceptable safety profile and provides a protective immune response against Ebola virus disease (EVD) induced by Zaire ebolavirus in humans. The purpose of this study was to determine whether the V920 vaccine is capable of replicating in arthropod cell cultures of relevant vector species and of replicating in live mosquitoes. While the V920 vaccine replicated well in Vero cells, no replication was observed in Anopheles or Aedes mosquito, Culicoides biting midge, or Lutzomyia sand fly cells, nor in live Culex or Aedes mosquitoes following exposure through intrathoracic inoculation or feeding on a high-titer infectious blood meal. The insect taxa selected for use in this study represent actual and potential epidemic vectors of VSV. V920 vaccine inoculated into Cx. quinquefasciatus and Ae. aegypti mosquitoes demonstrated persistence of replication-competent virus following inoculation, consistent with the recognized biological stability of the vaccine, but no evidence for active virus replication in live mosquitoes was observed. Following administration of an infectious blood meal to Ae. aegypti and Cx. quinquefasciatus mosquitoes at a titer several log10 PFU more concentrated than would be observed in vaccinated individuals, no infection or dissemination of V920 was observed in either mosquito species. In vitro and in vivo data gathered during this study support minimal risk of the vector-borne potential of the V920 vaccine.

Polyprotein Processing as a Determinant for in Vitro Activity of Semliki Forest Virus Replicase.

Semliki Forest virus (SFV) is an arthropod-borne alphavirus that induces membrane invaginations (spherules) in host cells. These harbor the viral replication complexes (RC) that synthesize viral RNA. Alphaviruses have four replicase or nonstructural proteins (nsPs), nsP1-4, expressed as polyprotein P1234. An early RC, which synthesizes minus-strand RNA, is formed by the polyprotein P123 and the polymerase nsP4. Further proteolytic cleavage results in a late RC consisting of nsP1-4 and synthesizing plus strands. Here, we show that only the late RCs are highly active in RNA synthesis in vitro. Furthermore, we demonstrate that active RCs can be isolated from both virus-infected cells and cells transfected with the wild-type replicase in combination with a plasmid expressing a template RNA. When an uncleavable polyprotein P123 and polymerase nsP4 were expressed together with a template, high levels of minus-strand RNA were produced in cells, but RCs isolated from these cells were hardly active in vitro. Furthermore, we observed that the uncleavable polyprotein P123 and polymerase nsP4, which have previously been shown to form spherules even in the absence of the template, did not replicate an exogenous template. Consequently, we hypothesize that the replicase proteins were sequestered in spherules and were no longer able to recruit a template.

New insights about ORF1 coding regions support the proposition of a new genus comprising arthropod viruses in the family Totiviridae.

Analyzing the positions of 2A-like polypeptide cleavage sites in all available genomes of arthropod totiviruses we propose the limits of all ORF1 coding sequences and observed that two proteins previously predicted in infectious myonecrosis virus genome are unique in the arthropod totiviruses group. A putative protein cleavage site upstream the major capsid protein was also identified only in these genomes. In addition, protein models generated using ab initio and threading approaches revealed conserved structures possibly related to formation of viral protrusions and RNA packaging, clarifying the mechanisms involved in the extracellular transmission. These data appoints that the group formed by arthropod totiviruses are sufficient distinctive to be clustered in new genus belonging to the Totiviridae family, in agreement with previous phylogenetic analysis.

Arthropod-Borne Diseases: The Camper's Uninvited Guests.

Arthropod-borne diseases are a major problem whenever outdoor activities bring arthropods and people into contact. The arthropods discussed here include arachnids (ticks) and insects. Most arthropod bites and stings are minor, with the notable exception being bee-sting anaphylaxis. Ticks cause the most disease transmission. Key hard tick vectors include black-legged (Ixodes), dog (Dermacentor), and lone star (Amblyomma) ticks, which transmit Lyme and various rickettsial diseases. Insect repellents, permethrin sprays, and proper tick inspection reduce this risk significantly. Lyme disease and the milder southern-tick-associated rash illness (STARI) are characterized by the erythema migrans rash followed, in the case of Lyme disease, by early, disseminated, and late systemic symptoms. Treatment is with doxycycline or ceftriaxone. Indefinite treatment of "chronic Lyme disease" based on subjective symptoms is not beneficial. Rickettsial diseases include ehrlichiosis, anaplasmosis, and Rocky Mountain spotted fever, which are characterized by fever, headache, and possible rash and should be empirically treated with doxycycline while awaiting laboratory confirmation. Tularemia is a bacterial disease (Francisella) spread by ticks and rabbits and characterized by fever and adenopathy. Treatment is with gentamicin or streptomycin. Babesiosis is a protozoal disease, mimicking malaria, that causes a self-limited flu-like disease in healthy hosts but can be life threatening with immune compromise. Treatment is with atovaquone and azithromycin. Other tick-related conditions include viral diseases (Powassan, Colorado tick fever, heartland virus), tick-borne relapsing fever (Borrelia), and tick paralysis (toxin). Mosquitoes, lice, fleas, and mites are notable for their annoying bites but are increasingly significant disease vectors even in the United States.

Complete genome sequences of Chandipura and Isfahan vesiculoviruses.

Chandipura virus (CHPV) and Isfahan virus (ISFV) are two members of the genus Vesiculovirus from Asia. Both are arthropod-transmitted and are able to infect humans, but neither causes vesicular stomatitis in livestock. The complete genome sequence for each virus has been determined. The negative-sense RNA genome comprises 11,119 nt (CHPV) or 11,088 nt (ISFV). The most variable of the non-transcribed regions is the intergenic spacer at the G-L gene junction (4 bases in ISFV, 20 in CHPV). Phylogenetic analysis of deduced protein sequences shows that although CHPV and ISFV are distinct viruses, they are more related to each other than either is to the New World vesicular stomatitis viruses (VSV). The South American virus, Piry virus, is more closely related to the Asian viruses ISFV and CHPV, than it is to VSV.