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Biomedical subjects

C Rouzioux

Publications and source records attributed to C Rouzioux.

At least 19 recordsLinked to original sources

Examination of real-time PCR for HIV-1 RNA and DNA quantitation in patients infected with HIV-1 BF intersubtype recombinant variants.

The impact of HIV-1 genetic diversity on the performance of laboratory testing is an issue that has to be monitored continuously. An "in-house" real-time PCR assay was developed by the Agence Nationale de Recherche sur le SIDA (ANRS) in France for viral load (VL) quantitation based on the amplification of the HIV-1 long terminal repeat (LTR) region. This technology has not been used in Argentina yet and considering the HIV-1 diversity in the country, a comparative analysis of this assay was undertaken versus the Versant HIV-1 RNA 3.0 Assay (b-DNA). The performance was assessed on 30 drug-naïve HIV-1 infected patients who were characterized previously by phylogenetic analysis of the pol and vpu gene. The results showed that there is a significant linear correlation between values of transformed viral load logarithms measured by both, bDNA and real-time PCR assay and that this assay can be used to quantify viral load in samples from BF-infected patients with the same accuracy and reliability as for B subtype samples. The use of "in-house" real-time PCR to measure DNA in PBMCs correlated strongly with the HIV-1 RNA levels in all specimens.

DNA, Viral↗

Increase in hepatitis C virus incidence in HIV-1-infected patients followed up since primary infection.

BACKGROUND: An increase in the incidence of sexually transmitted infections and hepatitis C virus (HCV) infections in HIV-infected men who have sex with men (MSM) has recently been reported. OBJECTIVE: To estimate HCV incidence and risk factors among HIV-1-infected patients followed up since primary HIV infection in the French PRIMO Cohort between 1996 and 2005. PATIENTS AND METHODS: All patients with at least 18 months of follow-up were studied. HCV antibody tests were performed on baseline plasma samples and repeated on the latest available sample when negative at baseline. RESULTS: In total, 402 patients with a median follow-up of 36 (range 18-104) months were eligible. HCV seroconversion was observed in 6 patients (4 men and 2 women), corresponding to an HCV incidence rate of 4.3 per 1000 person-years. Incidence rates in men and women were 3.5 and 7.8 per 1000 person-years, respectively. The incidence rate was 1.2 per 1000 person-years before January 2003 and 8.3 per 1000 person-years after January 2003 (p = 0.06). The classic risk factors for HCV infection were found in women (intravenous drug use, and body piercing), whereas the only identified risk factor for HCV acquisition was unsafe sex in the four men. CONCLUSIONS: Increase in the incidence of acute HCV infection in recently HIV-infected patients confirms the shift in sexual behaviour in the recent years, especially in HIV-infected MSM. Repeated testing for HCV antibodies should be carried out in HCV-negative HIV-infected patients and specific recommendations about protected sex should be clearly provided.

AIDS-Related Opportunistic Infections↗

Description of an adenovirus A31 outbreak in a paediatric haematology unit.

Adenovirus infections result in significant morbidity and mortality in allogeneic haematopoietic stem cell transplanted (hSCT) children. Adenovirus from species C and B account for more than 90% of adenoviruses recovered after hSCT. However, infections due to adenovirus A31 have been increasingly reported in recent years. Between April 2002 and April 2005, blood samples obtained every 2 weeks from 58 hSCT children were screened for adenovirus species A to C by quantitative real-time PCR. Phylogenetic analysis was realized after amplification and sequencing of the entire hexon gene. Fifteen cases of adenovirus infection with viraemia were recovered during this 3 years period. During spring/summer 2003, seven cases occurred and were due to an adenovirus species A. Phylogenetic analysis of the seven strains showed that they belonged to the A31 genotype and shared 100% homology. Clinical features of the seven HSCT children with A31 adenovirus viraemia are described. We describe here an epidemic spread of adenovirus genotype A31 in a paediatric haematology unit. Timing, location and hexon gene genotyping results highly suggested a nosocomial origin to this epidemic. The burden of adenovirus A31 infection needs to be further assessed in this context.

Adenovirus Infections, Human↗

Impact of highly active antiretroviral therapy on the maturation of anti-HIV-1 antibodies during primary HIV-1 infection.

OBJECTIVES: To study the impact of highly active antiretroviral therapy (HAART) on isotype switching and avidity maturation of HIV-1-specific immunoglobulin G (IgG) in patients with primary HIV-1 infection (PHI). METHODS: We studied the emergence and the evolution of anti-HIV IgG antibodies by quantitative immunoblotting to analyse IgG subclasses and IgG avidity. Serum samples were obtained from 16 PHI patients from the French PRIMO Cohort Study at various points in the first year of infection: eight patients received no treatment (group I), and eight patients received efficient HAART (group II) during the study period. RESULTS: Early initiation of HAART in PHI patients partially prevented an increase in anti-HIV-1 IgG levels. Within IgG subclasses, the amount of anti-HIV-1 IgG1 gradually increased with time in both groups, although levels remained lower in treated patients. The anti-p24 IgG2 level was always lower in group II. We observed a decrease in anti-p24 IgG3 over time in both groups. Treatment did not affect the maturation of HIV-1 IgG avidity, which increased in both groups until month 3 and then remained high until the end of the 12-month follow-up period. CONCLUSIONS: HAART in PHI partially prevents the emergence of HIV-1 IgG antibodies, but does not affect the quality of these antibodies, as reflected in their isotype and avidity.

Adolescent↗

Detailed analysis of the genetic evolution of influenza virus during the course of an epidemic.

The genetic variability of influenza virus is usually studied with sequences selected over numerous years and countries, and rarely within a single season. Here we examined the viral evolution and the correlation between genetic and clinical features during an epidemic. From a French prospective household-based study in 1999-2000, 99 infected patients were randomly selected. The HA1 genomic domain was sequenced. Phylogenetic analysis showed the existence of two groups of A/H3N2 viruses. We found no distinct pattern of genomic evolution within either group according to time. A spatial correlation with the nucleotide distances was shown. The average nucleotide diversity was 3.4x10-3 nucleotides per site, and did not differ between the groups. A lower number of segregating sites was observed in patients who experienced influenza-like symptoms during the previous epidemic. These results suggest that the influenza virus undergoes regular HA1 nucleotide changes, but without clonal expansion of mutant strains within a single epidemic.

Adult↗

The frequency of HIV-specific interferon- gamma -producing CD8 T cells is associated with both age and level of antigenic stimulation in HIV-1-infected children.

Ex vivo interferon (IFN)- gamma -producing CD8 T cells specific for human immunodeficiency virus (HIV) Env, Gag, and Pol antigens were measured in the peripheral blood of 55 children not receiving highly active antiretroviral therapy (HAART) and 70 children receiving HAART. In children not receiving HAART, the frequency of HIV-specific IFN- gamma -producing CD8 T cells was positively correlated with age and was not associated with plasma viral load or CD4 T cell levels. In children receiving HAART, the frequency of HIV-specific IFN- gamma -producing CD8 T cells was directly correlated with plasma viral load, and its association with age remained significant. In conclusion, the frequency of HIV-specific IFN- gamma -producing CD8 T cells in children is primarily determined by both age and plasma viral load.

Adolescent↗

Recent changes in the management of primary HIV-1 infection: results from the French PRIMO cohort.

OBJECTIVES: To describe the management of primary HIV infection (PHI), focusing on changes in the design of therapies and time to initiation of antiretroviral treatment, the clinical outcome, and the immuno-virological response over time to highly active antiretroviral therapy (HAART) and its tolerance. DESIGN AND METHODS: In the French PRIMO multicentre cohort, 291 patients presenting with PHI were enrolled between 1996 and 2001. Data were analysed to describe treatment prescription habits over a period of 5 years, and response to and tolerance of treatment. RESULTS: The proportion of patients who initiated treatment during PHI decreased from 92% in 1996 to 56% in 2001. At 6 months, whatever the initiated treatment, 74% of treated patients achieved a plasma viral load<400 HIV-1 RNA copies/mL and 53% achieved a viral load of<50 copies/mL. Prescription of protease inhibitor (PI)-sparing regimens has become more frequent since 1999. Despite a similar virological response, patients in the PI-containing group tended to experience a greater 1-year increase in CD4 cell count than those in the non-nucleoside reverse transcriptase (NNRTI)-containing group (218 cells/microL versus 157 cells/microL, respectively). An adverse event was recorded in 51% of treated patients. The most frequent events were gastrointestinal disorders (71%), lipodystrophy (27%) and mood disorders (19%). The main reason for modifying or stopping therapy was the occurrence of an adverse event. CONCLUSIONS: Limitations of therapy and poor tolerance to antiretroviral regimens have changed physician attitudes in PHI. This suggests the need for evaluation of better-tolerated regimens and new therapeutic strategies.

Adult↗

Field evaluation of an improved assay using a heat-dissociated p24 antigen for adults mainly infected with HIV-1 CRF02_AG strains in Côte d'Ivoire, West Africa.

The aim of this study was to evaluate the heat-dissociated p24 antigen (HD p24 Ag) assay as an alternative low-cost tool for diagnosis of HIV-1 infection and quantitation of HIV-1 RNA levels in African adults mainly infected with HIV-1 CRF02_AG strains. One hundred seventeen plasma specimens were obtained from HIV-1-seropositive subjects enrolled in the ANRS 1220 PRIMO-CI cohort (Abidjan, Côte d'Ivoire, West Africa). Results of the HD p24 Ag assay were comparable with those of HIV-1 RNA levels quantified in the same antibody-positive plasma samples by the Amplicor HIV-1 Monitor assay (1.5 version; Roche Diagnostics, Indianapolis, IN): sensitivity, 95.7% versus 96.6%, respectively; specificity (evaluated with samples from 75 seronegative subjects), 94.7% versus 100%, respectively. HD p24 Ag and HIV-1 RNA assays were weakly correlated (Spearman coefficient correlation, r = 0.33; P < 0.001) except for HIV-1 RNA levels of >/=5 log10 copies/mL (r = 0.62; P < 0.001). Quantitation of HD p24 antigenemia in 76 plasma specimens from 14 patients treated with highly active antiretroviral therapy demonstrated weaker changes during treatment than those observed with the HIV-1 RNA assay. Follow-up of infected patients using both markers showed different results. The reliability of the HD p24 Ag assay is questionable for clinical and biologic management as a surrogate tool for measurement of HIV-1 RNA levels in Africa.

Adolescent↗

[Management of the bank of biological material in the hospital-based SEROCO and HEMOCO cohorts of HIV-infected patients].

The French SEROCO and HEMOCO hospital-based cohorts have enrolled and followed HIV-infected patients, before and after the highly active antiretroviral therapy era. Among the objectives in 1988, was explicitly mentioned the constitution of a centralised bank of biological material (serums at enrollment and every 6 months, PBMC at enrollment and every 18 months). This paper details the organisation of the bank and the numerous studies performed from this bank, and presents a few simple decision rules which have been developed with the growing acquired experience.

Antiretroviral Therapy, Highly Active↗

[HIV and pregnancy].

HIV infection is a complex model of mother-to-child transmission. Antiretroviral therapy can induce a dramatic reduction of the risk of transmission, actually the rate of transmission is about 1 to 2 % in France. The actual most important stake concerns the diagnosis and the follow up of infected women living in countries of high endemy where the programs are inappropriate at the moment. The knowledge of the mechanisms and the timing of HIV mother-to-child transmission need to be more developed in order to design adapted antiretroviral posology and to try to reduce infant's exposure to drug toxicity.

Anti-HIV Agents↗

Immunological and virological features of HIV-infected patients with increasing CD4 cell numbers despite virological failure during protease inhibitor-based therapy.

OBJECTIVES: To investigate the extent of functional T cell recovery and to characterize plasma virus and virus producing cells in patients with increasing CD4 cell counts despite virological failure during protease inhibitor (PI) based therapy. METHODS: The study group included 13 patients who were treated for at least 12 months with a PI based regimen and were selected on the basis of a sustained immunological response (increase of > 70 CD4 cells/microL) despite virological failure (< 1 log10 copies/mL decrease in HIV-1 RNA plasma levels). RESULTS: Compared to a historical series of 11 complete responders with less advanced disease, the proportion of memory CD4 T cells was significantly higher (67.8+/-17.8 vs. 52.8+/-11.0; P=0.045) and the proportion of naive CD4 T cells significantly lower (30.5+/-14.8 vs. 45.0+/-10.4, P=0.021) in patients who were immunological responders/virological nonresponders. In those patients, ongoing viral replication was associated with a strong activation of circulating CD8 T lymphocytes; interleukin-2 production remained decreased. CD4 T cell reactivity to cytomegalovirus proteins was observed in nine of 11 patients tested. In the study group, the proportion of infectious virus present in plasma as well as the levels of intracellular viral replication were similar to those measured in untreated patients. Virological failure in this group of patients probably resulted from pre-existing mutations in the reverse transcriptase gene. CONCLUSIONS: This study of patients with increasing CD4 cell numbers despite virological failure shows the persistence of immune activation and partial immune restoration with no evidence of specific viral dynamics in vivo.

Adult↗

Semen alterations in HIV-1 infected men.

BACKGROUND: Couples in whom the man is infected by human immunodeficiency virus (HIV) increasingly request assisted reproductive technology (ART) to allow safe procreation. Semen quality is critical in such situations. METHODS: Semen characteristics were evaluated in 189 HIV-infected men requesting ART. At the time of semen analysis all men were healthy and 177 were receiving anti-retroviral therapy. Comparisons were made with HIV-seronegative men, partners of women requiring IVF because of tubal infertility, after matching for age and sexual abstinence delay. RESULTS: The most significant semen alterations found in the HIV-infected men were reduced percentages of rapidly progressive sperm [median (range), 10% (0-30%) compared with 15% (5-30%) in the controls, P < 0.001], and increased concentrations of non-spermatic cells [3 x 10(6)/ml (0.2-16 x 10(6)/ml) compared with 1.1 x 10(6)/ml (0.1-14 x 10(6)/ml) in the controls, P < 0.001]. HIV-infected men also showed lower ejaculate volumes [2.8 ml (0.6-9.3 ml) compared with 3.6 ml (1.1-11 ml), P < 0.05] and total sperm counts [262.5 x 10(6) (0-1003 x 10(6)) compared with 310.5 x 10(6) (48.3-1679 x 10(6)), P < 0.05]. CONCLUSIONS: Semen evaluation in a large population of HIV-infected men requesting ART evidenced several alterations. Some of these anomalies might be related to anti-retroviral treatments.

Adult↗

HIV type 1-specific IgG2 antibodies: markers of helper T cell type 1 response and prognostic marker of long-term nonprogression.

The helper T type 1 (Th1) function of CD4(+) T lymphocytes is presumed to be of key importance in host defense against HIV-1. As the production of different antibody isotypes is dependent on this helper T function, we investigated whether HIV-1-specific responses of a particular IgG isotype could be a reliable marker of long-term HIV-1 control. Assessment of the IgG subclass distribution in the plasma of HIV-1-infected patients enrolled in the French prospective Asymptomatic Long-Term (ALT) cohort showed that IgG2 directed against HIV-1 Env gp41 and Gag proteins was associated with low viral load, high CD4(+) lymphocyte count, and weak neutralizing activity. By contrast, levels of anti-Env and anti-Pol IgG1 as well as the magnitude of neutralizing activity were correlated with the viral load and thus merely reflect the level of HIV replication. Furthermore, IgG2 directed against Gag proteins was significantly associated with HIV-1 p24-specific Th1 cell production of interferon gamma and interleukin 2. In multivariate analysis, only two variables, anti-gp41 IgG2 and plasma HIV-1 RNA, were found to be independent prognostic factors of remaining long-term nonprogressive over time. By providing new insight into the nature of an HIV-specific antibody response associated with the control of virus replication, these findings have implications for the design of HIV vaccines.

Biomarkers↗

Early diagnosis of paediatric HIV-1 infection among African breast-fed children using a quantitative plasma HIV RNA assay.

OBJECTIVE: To evaluate the performance of a quantitative plasma HIV-1 RNA assay for HIV infection diagnosis among African breast-fed children. METHODS: Serial plasma specimens collected in the first week, at day 45-90, 6 months and 9-12 months of age from HIV-exposed children born to HIV-1-infected women enrolled in the DITRAME ANRS 049a perinatal intervention trial (Abidjan, Côte d'Ivoire) were tested for HIV-1 plasma RNA using a branched DNA (bDNA) assay. Sensitivity and specificity of this RNA test were assessed in comparison with a qualitative DNA polymerase chain reaction (PCR) performed on the same blood samples and allowing a reliable detection of the predominant subtype A. RESULTS: Among 91 samples from 53 infected children which tested positive by DNA PCR, the sensitivity of the bDNA test was 100% [95% confidence interval (CI), 96.0-100.0] at < or = 8 days (n = 19), 6-12 weeks (n = 43), 6 months (n = 26), and 9-12 months (n = 3). The median plasma HIV-1 RNA viral load ranged from 242 000 copies/ml at < or = 8 days to more than 500 000 copies/ml at day 45-90 and at 6 months. Of 106 specimens from 106 uninfected children who were DNA PCR- negative at month 3 or 6 of age, HIV-1 RNA was undetectable in 103, yielding an overall specificity for the bDNA test of 97.2% (95% CI, 92.0-99.4). The viral load in the three remaining samples with false-positive results was low (410, 937 and 3752 copies/ml, respectively). CONCLUSIONS: The quantitative bDNA assay appears a suitable tool for early, reliable and easy diagnosis of paediatric HIV-1 infection among a population of African breast-fed children.

Africa↗

Immunological and virological effects of long term IL-2 therapy in HIV-1-infected patients.

We report the long-term outcome of 27 HIV-infected patients treated for over 3 years with IL-2 and binucleoside analogues. These patients experienced a sustained increase in CD4 cells and a decrease of proviral DNA with infrequent IL-2 cycles. In three cases, virus could not be isolated from activated peripheral cells. A high frequency of HIV-1-specific memory CD4 T cells was found in the patients studied. IL-2 maintains specific effector cells and reduces the pool of infected cells in patients, albeit treated only with binucleosides.

Anti-HIV Agents↗

Early control of HIV replication in primary HIV-1 infection treated with antiretroviral drugs and pegylated IFN alpha: results from the Primoferon A (ANRS 086) Study.

IFN alpha has both antiviral and immunostimulating properties. The ANRS086 Primoferon A Study evaluated in 12 patients with primary HIV infection the tolerance and efficacy of an early and transient administration of pegylated IFN alpha, in addition to highly active antiretroviral therapy. Tolerance was good, and this regimen allowed the early control of HIV replication and rapid decay of the viral reservoir. These results support the initiation of comparative studies with pegylated INF alpha in primary HIV infection.

Antiretroviral Therapy, Highly Active↗

Zidovudine genotypic resistance in HIV-1-infected newborns in the French perinatal cohort.

A retrospective study was set up to investigate the frequency of zidovudine (ZDV)-resistant HIV-1 in infected newborns after ZDV prophylaxis in the French Perinatal Cohort study. Nucleotide sequence analysis was carried out from 34 infants' isolates and 18 maternal plasma samples. Mutations related to ZDV resistance were found in the HIV-1 reverse transcriptase in 7 of 34 children (20%). Evidence of mother-child transmission of ZDV-resistant HIV-1 was found in 4 cases. Phylogenetic analysis showed that 14 of 34 HIV-1 isolates from the infants belonged to non-B subtypes. The presence of ZDV resistance-encoding mutations in the newborn isolates was associated with a longer total duration of exposure to ZDV. In a context of a wide HIV-1 variability, ZDV resistance can be one of the factors contributing to mother-child transmission.

Anti-HIV Agents↗