Characterization of the Molecular Determinants of Primary HIV-1 Vpr Proteins: Impact of the Q65R and R77Q Substitutions on Vpr Functions

Although HIV-1 Vpr displays several functions in vitro, limited information exists concerning their relevance during infection. Here, we characterized Vpr variants isolated from a rapid and a long-term non-progressor (LTNP). Interestingly, vpr alleles isolated from longitudinal samples of the LTNP revealed a dominant sequence that subsequently led to diversity similar to that observed in the progressor patient. Most of primary Vpr proteins accumulated at the nuclear envelope and interacted with host-cell partners of Vpr. They displayed cytostatic and proapoptotic activities, although a LTNP allele, harboring the Q65R substitution, failed to bind the DCAF1 subunit of the Cul4a/DDB1 E3 ligase and was inactive. This Q65R substitution correlated with impairment of Vpr docking at the nuclear envelope, raising the possibility of a functional link between this property and the Vpr cytostatic activity. In contradiction with published results, the R77Q substitution, found in LTNP alleles, did not influence Vpr proapoptotic activity

Simple scoring system to predict in-hospital mortality after surgery for infective endocarditis

BACKGROUND: Aspecific scoring systems are used to predict the risk of death postsurgery in patients with infective endocarditis (IE). The purpose of the present study was both to analyze the risk factors for in-hospital death, which complicates surgery for IE, and to create a mortality risk score based on the results of this analysis. METHODS AND RESULTS: Outcomes of 361 consecutive patients (mean age, 59.1\ub115.4 years) who had undergone surgery for IE in 8 European centers of cardiac surgery were recorded prospectively, and a risk factor analysis (multivariable logistic regression) for in-hospital death was performed. The discriminatory power of a new predictive scoring system was assessed with the receiver operating characteristic curve analysis. Score validation procedures were carried out. Fifty-six (15.5%) patients died postsurgery. BMI >27 kg/m2 (odds ratio [OR], 1.79; P=0.049), estimated glomerular filtration rate 55 mm Hg (OR, 1.78; P=0.032), and critical state (OR, 2.37; P=0.017) were independent predictors of in-hospital death. A scoring system was devised to predict in-hospital death postsurgery for IE (area under the receiver operating characteristic curve, 0.780; 95% CI, 0.734-0.822). The score performed better than 5 of 6 scoring systems for in-hospital death after cardiac surgery that were considered. CONCLUSIONS: A simple scoring system based on risk factors for in-hospital death was specifically created to predict mortality risk postsurgery in patients with IE

Acceptabilité et faisabilité d'un dépistage systématique du VIH en médecine générale

PARIS6-Bibl. St Antoine CHU (751122104) / SudocSudocFranceF

Evaluation de la procédure de prise en charge des accidents d'exposition au VIH au CHU d'Amiens (à propos de 628 dossiers sur deux années (2001-2002))

Un Accident d'Exposition au Sang ou aux Sécrétions (AES) est défini par un contact avec du sang ou un liquide contenant du sang possiblement infecté par le virus de l'immunodéficience humaine (VIH), mais aussi par les virus des hépatites (VHB ou VHC). Lors de ces accidents, qu'ils soient professionnels en milieu de soins ou qu'ils surviennent lors de rapports sexuels à risque, ou chez les usagers de drogue par voie intraveineuse, il existe un risque quantifiable de contamination virale. Nous avons voulu étudier, à la manière d'un audit, l'ensemble des moyens mis en œuvre au CHU d'Amiens pour la prise en charge de ces accidents très spécifiques. Nous avons repris pour cela les dossiers des 628 consultants pour AES au CHU d'Amiens sur 2001 et 2002. Il s'agit là d'une prise en charge complexe et pluridisciplinaire qui nécessite la mise au point de procédures de travail et de protocoles accessibles à tous. Une évaluation régulière des pratiques reste nécessaire au sein du CHUAMIENS-BU Santé (800212102) / SudocPARIS-BIUM (751062103) / SudocSudocFranceF

Incidence of Herpes Zoster in HIV-Infected Adults in the Combined Antiretroviral Treatment (cART) Era: Results from the FHDH-ANRS CO4 Cohort

International audienceBackground: Recent studies have shown a decrease in the incidence of herpes zoster (HZ) among HIV-infected patients since the cART era, but more data are needed on a possible increase in the risk early after cART initiation.Methods: We studied the HZ incidence and risk factors among patients followed in the French Hospital Database on HIV (FHDH) between 1992 and 2011. Standardized incidence ratios (SIR) were used for comparison with the general population between 2005 and 2008. The risk of HZ following cART initiation (0-6, ≥6 months) was studied with Poisson regression models.Results: 7167 cases of incident HZ were diagnosed among 91 044 individuals (583 125 person-years). The incidence declined significantly from 2955 per 100 000 person-years in 1992-1996 to 628 in 2009-2011. This decline was mainly explained by cART (RR=0.60; 95%CI, 0.57-0.64). The risk of HZ was associated with low CD4 cell counts, high HIV-RNA levels, low CD4/CD8 ratios and prior AIDS. Compared to the general population, the risk of HZ was higher in HIV-infected patients (overall SIR=2.7; 95%CI, 2.6-2.9), particularly between ages 15 and 45 years (SIR=4-6). In ART-naive patients a moderate increase in the HZ risk was observed during the first 6 months of cART, with a peak at 3 months (RR=1.47 95%CI, 1.26-1.73) a finding that disappeared after adjustment for the current CD4 cell count (RR=1.03; 95%CI, 0.81-1.32). Conclusions: The risk of HZ has declined markedly among HIV-infected patients in the cART era but remains 3 times higher than in the general population. The risk increases moderately during the first 6 months of cART

Interaction of the Vpr proteins from the LTNP and PR patients with hCG1, UNG2 and DCAF1 in the yeast two-hybrid system.

<p>The L40 reporter yeast strain expressing Vpr<i>Lai</i> or the indicated primary Vpr proteins from the LTNP and PR patients fused to LexA (LexA hybrid), in combination with either hCG1, UNG2 or DCAF1 fused to the Gal4 activation domain (Gal4AD hybrid), was analyzed for histidine auxotrophy. Growth in the absence of histidine indicates interaction between hybrid proteins.</p

G2-arrest and pro-apoptotic activities of Vpr proteins from the LTNP and PR patients.

<p>HPB-ALL T lymphoid cells were co-transfected with vectors for expression of HA tagged Vpr<i>Lai</i> or the indicated primary Vpr proteins from the LTNP and PR patients, in combination with the GFP expression vector. A) Cellular expression of HA-tagged Vpr proteins. Lysates from HPB-ALL transfected cells were analyzed by western-blotting using anti-GFP and anti-HA antibodies. B) G2-arrest activity of primary Vpr proteins. 48 h after transfection, cells were fixed and the DNA content of GFP-positive cells was analyzed by flow cytometry after DNA staining with propidium iodide. Results are expressed as the G2M/G1 ratios relative to that of the VprLai. C) Pro-apoptotic activity of Vpr proteins. Cells co-expressing the GFP and HA-tagged Vpr proteins were assayed by flow cytometry 72 h following transfection for cell surface phosphatidylserine exposure using AnnexinV coupled to phycoerythrin. Values are means of three independent experiments; error bars represent one standard deviation from the mean.</p

Description of patients and <i>vpr</i> alleles analyzed in the study.

<p>A) Immunological and viral profiles of the patients analyzed. These patients were selected from the HIV-1-infected patient cohort of the St-Antoine Hospital (Paris), and the samples were collected with written informed consent <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007514#pone.0007514-Lefrere1" target="_blank">[45]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007514#pone.0007514-Lefrere2" target="_blank">[46]</a>. The graphs show the time-course evolution of the blood CD4+ T cell counts (green curves) and plasmatic virus load (viral RNA, red curves); the PBMC samples analyzed in the present study are indicated by the blue arrows. B) Alignment of the amino acid sequences derived from DNA sequencing of the <i>vpr</i> alleles cloned from PBMC DNA samples. Excepted for LTNP-05 and LTNP-07 sequences that were obtained by direct sequencing of the bulk PCR fragments amplified from these samples, at least 40 independent clones were sequenced from each other samples. The sequences of the primary Vpr proteins are aligned with respect of the prototypic HIV-1<i>Lai</i> sequence (upper sequence). The <i>vpr</i> alleles from the LNTP (patient 5071) that were selected for subsequent functional analysis are in the box; the R77Q substitution identified in some <i>vpr</i> alleles is indicated in blue, while the Q65R substitution identified in the Vpr LTNP-04-2 is indicated in red.</p

Impact of the residue 77 on pro-apoptotic and G2-arrest activities of Vpr LTNP variants.

<p>The Arg77 residue from VprLai and Vpr LTNP-96 was replaced by a Gln, whereas the Gln residue from LTNP-86 and LTNP-04-1 was replaced by an Arg. The wild-type (grey bars) and mutated (hatched bars) Vpr proteins were then analyzed for apoptosis (A) and G2-arrest (B) activities as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007514#pone-0007514-g004" target="_blank">Figure 4</a>.</p

Relative frequency of primary Vpr alleles^a.

a<p><i>vpr</i> genes were amplified by PCR from each PBMC sample (indicated by blue arrows in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007514#pone-0007514-g001" target="_blank">Fig. 1A</a>), cloned into a shuttle plasmid, and DNA sequences from at least 40 independent clones were determined from each sample.</p>b<p>samples 1986, 1996 and 2004 are from the LTNP patient (5071), and sample PR is from the progressor patient (1200).</p>c<p>Vpr alleles correspond to the primary amino acid sequences reported in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007514#pone-0007514-g001" target="_blank">Fig. 1B</a>.</p