Tick-borne encephalitis virus (TBEV) is the most important tick-transmitted pathogen. It belongs to the
Flaviviridae
family and causes severe human neuroinfections. In this study, protective efficacy of the chimeric antibody chFVN145 was examined in mice infected with strains belonging to the Far-Eastern, European, and Siberian subtypes of TBEV, and the antibody showed clear therapeutic efficacy when it was administered once one, two, or three days after infection. The efficacy was independent of the TBEV strain used to infect the mice; however, the survival rate of the mice was dependent on the dose of TBEV and of the antibody. No enhancement of TBEV infection was observed when the mice were treated with non-protective doses of chFVN145. Using a panel of recombinant fragments of the TBEV glycoprotein E, the neutralizing epitope for chFVN145 was localized in domain III of the TBEV glycoprotein E, in a region between amino acid residues 301 and 359. In addition, three potential sites responsible for binding with chFVN145 were determined using peptide phage display libraries, and 3D modeling demonstrated that the sites do not contact the fusion loop and, hence, their binding with chFVN145 does not result in increased attachment of TBEV to target cells.
bMost Lyme borreliosis cases in Russia result from Borrelia garinii NT29 group infection. Borrelias of this group circulate exclusively in Ixodes persulcatus ticks, which are seldom found beyond Russia and the far east. Here we report the whole-genome sequence of Borrelia garinii BgVir isolated from an I. persulcatus female. L yme disease, caused by bacteria of the Borrelia genus, is the most frequent tick-borne infection in Russia (4). More than a million tick bites of humans were reported in the region last year. About 50% of the ticks investigated were infected with Borrelia, primarily of the B. garinii NT29 group (1, 2, 7). Borrelia bacteria of this group not only present a peculiar clinical picture, common in Russian patients, but are also limited in their inhabitation to Russia and northern China. These are also the only Borrelia bacteria that can effectively circulate in Ixodes persulcatus (5, 6). Hence, they are often omitted from the most prominent investigations, which usually rely on Borrelia strains common in Europe and North America. Here we report the first whole-genome sequence of a B. garinii NT29 group (strain BgVir).BgVir was isolated from an I. persulcatus female (2). Borrelia cells from the fourth passage on BSK-H media were used to avoid possible plasmid diminution. A rapid fragment library was sequenced on a Roche FLX instrument following the Titanium protocol, resulting in approximately 236 Mb of raw data with a mean read length of 341 nucleotides. A number of subsets of data, differing in coverage and mean read length, were then generated and fed to Newbler v. 2.3 to chase the best contig metrics. A 2ϫ 50-bp mate-paired library with a mean insertion size of 1 kb was additionally sequenced following the ABI SOLiD v.3.5. protocol. To achieve as narrow an insert size distribution as possible, the DNA shearing was followed by additional size selection on an agarose gel. The introduction of different sequencing platforms and library types allowed us to perform scaffolding and error correction in Roche FLX reads, which especially occurred in homopolymer stretches ubiquitous in the Borrelia genome. Gap sizes were derived from pairing of SOLiD reads. Local mapping of FLX reads was performed to the ends of each gap, followed by insertion of trimmed unmapped sequences. This operation was repeated until all gaps were closed. Validation and error correction were performed by mapping both the FLX and SOLiD reads on the obtained draft sequence. Full-length sequences of chromosome, linear plasmid lp54, and circular plasmid cp26 were obtained. The rest of the plasmids are represented by 46 scaffolds. The nucleotide sequences were annotated using GeneMark.hmm-PS and Glimmer algorithms.After alignment was performed, the chromosome nucleotide sequences of B. garinii BgVir and B. garinii PBi exhibited 98% identity. Most of the single nucleotide polymorphisms (SNPs) detected were synonymous polymorphisms. The most dramatic structural alteration is the insertion into BgVir of the chromosome sequence harboring ...
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