Novel approach for genotyping varicella-zoster virus strains from Germany

In this study, we present a novel genotyping scheme to classify German wild-type varicella-zoster virus (VZV) strains and to differentiate them from the Oka vaccine strain (genotype B). This approach is based on analysis of four loci in open reading frames (ORFs) 51 to 58, encompassing a total length of 1,990 bp. The new genotyping scheme produced identical clusters in phylogenetic analyses compared to full-genome sequences from well-characterized VZV strains. Based on genotype A, D, B, and C reference strains, a dichotomous identification key (DIK) was developed and applied for VZV strains obtained from vesicle fluid and liquor samples originating from 42 patients suffering from varicella or zoster between 2003 and 2006. Sequencing of regions in ORFs 51, 52, 53, 56, 57, and 58 identified 18 single-nucleotide polymorphisms (SNPs), including two novel ones, SNP 89727 and SNP 92792 in ORF51 and ORF52, respectively. The DIK as well as phylogenetic analysis by Bayesian inference showed that 14 VZV strains belonged to genotype A, and 28 VZV strains were classified as genotype D. Neither Japanese (vaccine)-like B strains nor recombinant-like C strains were found within the samples from Germany. The novel genotyping scheme and the DIK were demonstrated to be practical and simple and allow the highly efficient replication of phylogenetic patterns in VZV initially derived from full-genome DNA sequence analyses. Therefore, this approach may allow us to draw a more comprehensive picture of wild-type VZV strains circulating in Germany and Central Europe by high-throughput procedures in the future.

Tracking virus-specific CD4+ T cells during and after acute hepatitis C virus infection

BACKGROUND: CD4+ T cell help is critical in maintaining antiviral immune responses and such help has been shown to be sustained in acute resolving hepatitis C. In contrast, in evolving chronic hepatitis C CD4+ T cell helper responses appear to be absent or short-lived, using functional assays. METHODOLOGY/PRINCIPAL FINDINGS: Here we used a novel HLA-DR1 tetramer containing a highly targeted CD4+ T cell epitope from the hepatitis C virus non-structural protein 4 to track number and phenotype of hepatitis C virus specific CD4+ T cells in a cohort of seven HLA-DR1 positive patients with acute hepatitis C in comparison to patients with chronic or resolved hepatitis C. We observed peptide-specific T cells in all seven patients with acute hepatitis C regardless of outcome at frequencies up to 0.65% of CD4+ T cells. Among patients who transiently controlled virus replication we observed loss of function, and/or physical deletion of tetramer+ CD4+ T cells before viral recrudescence. In some patients with chronic hepatitis C very low numbers of tetramer+ cells were detectable in peripheral blood, compared to robust responses detected in spontaneous resolvers. Importantly we did not observe escape mutations in this key CD4+ T cell epitope in patients with evolving chronic hepatitis C. CONCLUSIONS/SIGNIFICANCE: During acute hepatitis C a CD4+ T cell response against this epitope is readily induced in most, if not all, HLA-DR1+ patients. This antiviral T cell population becomes functionally impaired or is deleted early in the course of disease in those where viremia persists.

1.0-M gadobutrol versus 0.5-M gadoterate for peripheral magnetic resonance angiography: a prospective randomized controlled clinical trial

PURPOSE: To perform a quantitative and qualitative comparison of gadobutrol and gadoterate in three-station contrast enhanced magnetic resonance angiography (CE-MRA) of the lower limbs. MATERIALS AND METHODS: In this prospective randomized controlled trial, 52 patients with leg ischemia were randomly assigned to one of two groups receiving either gadobutrol (1.0 mmol Gd/mL, 15 mL) or gadoterate (0.5 mmol Gd/mL, 30 mL). Three-station 3D CE-MRAs from the pelvis to the ankles were performed with moving-table technique on a 1.5T MR scanner. Injection time was identical in both groups. Signal-to-noise (SNR) and contrast-to-noise ratios (CNR) were calculated for 816 arteries. Contrast quality in 1196 vessel segments was evaluated separately by two blinded readers on a three-point scale. RESULTS: Mean SNR (61.8 +/- 7.8 for gadobutrol vs. 61.9 +/- 9.1 for gadoterate, P = 0.257), CNR (52.8 +/- 9.1 vs. 52.8 +/- 10.7, P = 0.154), and qualitative ranking (1.41 vs. 1.44, P = 0.21) for all vessels did not differ significantly between the two patient groups. The overall quality was good in 90.4% with gadoterate and 94.2% with gadobutrol (P = 0.462). CONCLUSION: High-concentration gadobutrol allows neither a higher CNR nor any qualitative advantage over the ordinary unspecific Gd agent gadoterate when the same Gd load and injection times are used in multistation CE-MRA of the peripheral arteries.