ABSTRACT
A new densovirus was isolated and characterized in laboratory strains of Toxorhynchites splendens. The virus was detected by polymerase chain reaction (PCR) from mosquitoes reared in our laboratory. PCR fragments from each mosquito were compared by single strand conformation polymorphism (SSCP) assay and found to be indistinguishable. Thus, it is likely the densoviruses from these mosquitoes contain homologous nucleotide sequences. The PCR fragment corresponding to a 451 bp densovirus structural gene segment from each of 5 mosquitoes had 100% identical nucleotide sequences. Phylogenetic analysis of the structural gene sequence suggests the newly isolated densovirus is more closely related to Aedes aegypti densovirus (AaeDNV) than to Aedes albopictus densovirus (AalDNV). Analysis of offspring and predated larvae suggests that vertical and horizontal transmission are responsible for chronic infections in this laboratory strain of Toxorhynchites splendens. The virion DNA is 4.2 kb in size, is closely related to, but distinct from, known densoviruses in the genera Brevidensovirus and Contravirus. Thevirus is tentatively named Toxorhynchites splendens densovirus (TsDNV).
Subject(s)
Culicidae/virology , Densovirus/genetics , Animals , Culicidae/classification , DNA, Viral/genetics , Dengue/parasitology , Dengue/transmission , Densovirus/classification , Densovirus/isolation & purification , Humans , Larva , Likelihood Functions , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , ThailandABSTRACT
BACKGROUND: Vascular leakage and shock are the major causes of death in patients with dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Thirty years ago, complement activation was proposed to be a key underlying event, but the cause of complement activation has remained unknown. METHODS: The major nonstructural dengue virus (DV) protein NS1 was tested for its capacity to activate human complement in its membrane-associated and soluble forms. Plasma samples from 163 patients with DV infection and from 19 patients with other febrile illnesses were prospectively analyzed for viral load and for levels of NS1 and complement-activation products. Blood and pleural fluids from 9 patients with DSS were also analyzed. RESULTS: Soluble NS1 activated complement to completion, and activation was enhanced by polyclonal and monoclonal antibodies against NS1. Complement was also activated by cell-associated NS1 in the presence of specific antibodies. Plasma levels of NS1 and terminal SC5b-9 complexes correlated with disease severity. Large amounts of NS1, complement anaphylatoxin C5a, and the terminal complement complex SC5b-9 were present in pleural fluids from patients with DSS. CONCLUSIONS: Complement activation mediated by NS1 leads to local and systemic generation of anaphylatoxins and SC5b-9, which may contribute to the pathogenesis of the vascular leakage that occurs in patients with DHF/DSS.
