[Assessment of the diagnostic value of RT-PCR on amniotic fluid for prenatal diagnosis of congenital rubella infection]. / L'infection rubéolique maternofoetale: apport de la biologie moléculaire.

AIM OF THE STUDY: To assess the diagnostic value of RT-PCR on amniotic fluid (AF) for prenatal diagnosis of congenital rubella infection. MATERIAL AND METHODS: RT-PCR on AF was compared to specific IgM antibody detection in foetuses and/or newborns in 45 pregnant women with confirmed primary infection. RESULTS: specificity of RT-PCR was 100% and sensitivity ranged between 83 and 95%. CONCLUSION: RT PCR may be considered as a valuable tool for prenatal diagnosis of foetal rubella infection.

[Prevalence of asymptomatic sexually transmitted infections among high-risk patients attending a free anonymous HIV-screening center]. / Prévalence des infections sexuellement transmissibles asymptomatiques chez des sujets à haut risque consultant dans un centre de dépistage anonyme et gratuit du SIDA.

INTRODUCTION: The aim of this study was to assess the prevalence of sexually transmitted infections among patients attending an anonymous HIV Screening Center. PATIENTS AND METHODS: This prospective study was performed in the HIV Screening Center of University hospital in Reims (France) from May 1997 to December 1997. The inclusion criteria were the asymptomatic clinical presentation and the presence of risk factors for sexually transmitted infections referring to WHO criteria. The methods included clinical examination after application of acetic acid and urethral and endocervical swabs to identify:Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis, Gardnerella vaginalis, Trichomonas vaginalis in specific culture. Treponema pallidum and HIV-1 infection were both detected by Enzym Linked Immuno Sorbent Assay (ELISA). RESULTS: One hundred and one patients (62 men and 39 women) were included in the study. Their mean age was 27 +/- 4 Years. Risk factors for sexually transmitted infections were: multiple sexual partners 81 p. 100; homo or bisexuality 16 p. 100; intravenous drug use 3 p. 100. The sexually transmitted infections were: HIV-1 infection 1 p. 100;Ureaplasma urealyticum 25 p. 100; genital warts 5 p. 100;Chlamydia trachomatis 3 p. 100; Gardnerella vaginalis 3 p. 100; Mycoplasma hominis 2 p. 100; Treponema pallidum 0 p. 100; Neisseria gonorrhoeae 0 p. 100; Trichomonas vaginalis 0 p. 100. The prevalence of sexually transmitted infections was significantly higher among women (p<0.05). DISCUSSION: Classical sexually transmitted infections and HIV infection were rarely detected in this study; but prevalence of other sexually transmitted infections (genital warts, Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum) was high. Ureaplasma urealyticum is considered as a possible pathogenic agent in pregnant women (preterm delivery, decrease of birth weight, chorioamniotitis). These results suggest that other than sexually transmitted infections in high risk patients attending a HIV Screening Center other sexually transmitted infections should also be systematically screened for.

Preferential and persistent depletion of CCR5+ T-helper lymphocytes with nonlymphoid homing potential despite early treatment of primary HIV infection.

Strains of human immunodeficiency virus (HIV) transmitted between individuals use the CCR5 coreceptor, but no preferential depletion of particular Th-lymphocyte subpopulations has been reported during primary HIV infection (PHI). In contrast, gut-associated Th lymphocytes are preferentially depleted in macaques recently infected by simian immunodeficiency virus. The expression of CCR5 and the intestinal homing receptor integrin alpha4beta7 on subpopulations of Th lymphocytes was studied in 12 patients with PHI. There was a profound decrease of circulating alpha4beta7+ Th lymphocytes and CCR5+ memory Th lymphocytes with nonlymphoid homing potential (CD62L-CD45RO+). Unlike other Th lymphocytes, this cell population remained depleted despite early control of viral replication under antiretroviral treatment. Therefore, HIV preferentially targets a specific CCR5+ subpopulation of Th lymphocytes early during infection, inducing its persistent depletion despite treatment. Protective immunity in vivo depends on Th lymphocytes carrying homing capacity to nonlymphoid tissue, and therefore these data may explain the persistent abnormalities of immune functions in patients infected with HIV.

[Sarcoidosis and progressive multifocal leukoencephalopathy]. / Sarcoïdose et leucoencéphalopathie multifocale progressive.

A 70-year-old woman treated for sarcoidosis complained of progressive cognitive impairment and gait disability. Magnetic resonance imaging of the brain revealed a nonenhancing lesion in T1-weighted imaging in the left parieto-occipital region and sarcoidosis of the central nervous system was evoked. However, she rapidly deteriorated with posterior and cerebellar extension of the lesions, suggesting of progressive multifocal leukoencephalopathy (PML). DNA of the JC virus (JCV) was detected in the cerebrospinal fluid (CSF) by a polymerase chain reaction. Despite antiviral therapy, she died nine months after the first neurological signs. This case illustrates the possible association between sarcoidosis and PML, and underlines the interest to detect the presence of JCV in the CSF when the diagnosis of neurosarcoidosis appeared uncertain.

Virological and histological responses to one year alpha-interferon-2a in hemodialyzed patients with chronic hepatitis C.

BACKGROUND: alpha-Interferon-2a (IFNalpha) alone is a therapy of limited proven benefit for non-uremic patients with chronic hepatitis C virus (HCV) infection. In dialyzed patients, such an effect is suggested on small short-term studies without sufficient clinical and virologic follow-up to document any sustained effect. PROTOCOL: Twelve chronically hemodialyzed patients with chronic hepatitis C and waiting for renal transplantation were included in a prospective open study of treatment with IFNalpha. We used, as did others, doses of 3 million units (MU), three times a week, but for a longer period of treatment of 12 months. Follow-up was continued for 6 months after the end of IFNalpha in order to document any sustained biochemical, virological and histological responses. RESULTS: Aminotransferase levels returned to the normal range within 1-2 months of treatment in all patients in whom they had been elevated at baseline. At 1 month of treatment, serum HCV-RNA was not detected in 5 (41%) patients and in 9 (75%) at 12 months. A sustained virological response was documented in 4 (33%) patients 6 months after the end of treatment. Relapse occurred in 5 patients within 2 months after IFNalpha withdrawal. HCV genotype was not predictive of any sustained response. At inclusion, using the histologic Metavir scoring system, half of the patients had low-grade cytolytic activity and none had cirrhosis. After IFNalpha, liver biopsy specimens were available from 9 patients and showed histologic improvement in 3. IFNalpha tolerance was poor, inducing a 5% mean weight loss and the acute rejection of two nonfunctioning kidney grafts. CONCLUSION: This study documents that administration of IFNalpha at 3 MU three times a week, for 12 months, in hemodialysis patients with chronic hepatitis C was efficient for clearing the serum of HCV-RNA in 75% of the patients. A sustained response was maintained in one third of these patients after cessation of IFNalpha, and was predicted by the early serum clearance of the virus within the first 2 months of treatment. We confirm that a 12-month treatment period carries a higher sustained response rate than shorter treatment periods. These encouraging results call for larger studies in uremic patients, using IFNalpha alone or in association with new antiviral drugs.

JC virus genotypes in France: molecular epidemiology and potential significance for progressive multifocal leukoencephalopathy.

JC virus (JCV) induces progressive multifocal leukoencephalopathy (PML), especially in human immunodeficiency virus (HIV)-infected patients. Although JCV genotypes have primarily been associated with geographic patterns, a distinctive neuropathogenicity was recently attributed to genotype 2. A multicenter study was conducted to describe the distribution of JCV genotypes in France and to investigate correlations between genotypes and PML. Genotypes were determined by sequencing 494 bp in the VP1 capsid gene. Peripheral JCV was studied in 65 urine samples from 43 HIV-infected patients and from 22 control subjects. Genotypes 1, 4, 2, and 3 were detected in 52.3%, 30.8%, 12.3%, and 4.6% of the samples, respectively. In 56 brain or cerebrospinal fluid samples, PML-associated JCV of genotypes 1, 2, 4, and 3 was found in 66%, 19.7%, 8.9%, and 5.4%, respectively. Infection with JCV genotypes 1 or 2 was correlated with PML (odds ratio, 3.29). On the other hand, infection with JCV genotype 4 could represent a lower risk for PML.

Isolation of an unusual Mycobacterium species from an AIDS patient with acute lymphadenitis.

A nonchromogenic Mycobacterium species was isolated from an AIDS patient with acute lymphadenitis. On the basis of the results of conventional tests, the strain appeared to be an atypical nonphotochromogenic Mycobacterium kansasii strain. Sequencing of the 16S rRNA gene revealed a unique nucleic acid sequence, suggesting that the isolate represents an undescribed pathogenic species.

[Anti-herpes chemotherapy]. / Chimiothérapie antiherpétique.

With the increasing number of immunocompromised patients over the last two decades, disease pattern caused by Herpesviridae has changed. More virulent, more severe, herpesvirus diseases are more frequently treated and consequently the drug-resistant herpesvirus mutants have arisen in the clinic. All these events justify to explore future directions in drug development and herpesviral research as antisens strategy or immunotherapy.

Prevalence of JC virus viraemia in HIV-infected patients with or without neurological disorders: a prospective study.

Progressive Multifocal Leukoencephalopathy (PML) is a severe demyelinating disease, which is rapidly fatal and is due to JC virus (JCV) infection, which especially occurs in HIV-infected patients. To investigate JCV pathophysiology and to evaluate the predictive value of JCV detection in blood, we looked for JCV DNA in leukocytes and plasma of 96 patients without any neurological symptoms and 109 patients with neurological diseases, among whom 19 were suffering from PML. JCV genome was detected in about 18% of all patients, i.e. 15.6% of patients with central nervous system disorders except PML, 13.5% of patients without neurological symptoms and significantly more often in PML patients (47.6%). Both leukocytes and plasma were tested; in plasma, JCV DNA was found in 36.1% of positive patients and in cells in 80.5%. Surprisingly in seven instances only the plasma contained JCV genome. One-year follow-up of these patients showed that the absence of JCV DNA in blood was associated with a very low probability of developing PML (negative predictive value=0.99).

[Progressive multifocal leukoencephalopathy: virological and neuropathological aspects]. / Leucoencéphalite multifocale progressive: aspects virologiques et neuropathologiques.

Progressive multifocal leukoencephalopathy is a subacute demyelinating disease of the central nervous system due to an opportunistic infection by a polyomavirus, most often JC virus, which predominantly infects oligodendrocytes. Progressive multifocal leukoencephalopathy used to be a rare condition, usually complicating lymphoproliferative diseases. Since the onset of the AIDS epidemic, its incidence has considerably increased and HIV infection has become, by far, the main risk factor for the disease. In AIDS patients, progressive leukoencephalopathy frequently shows atypical clinical and pathological features. The development of malignant glial tumors, within demyelinating regions, in patients with progressive multifocal leukoencephalopathy, has been reported in exceptional cases. The course of progressive multifocal leukoencephalopathy is invariably fatal. The diagnosis can only be made with certainty by histopathological examination of the brain, on cerebral biopsy or at postmortem. However, neuroradiological features may be extremely suggestive in many cases and PCR seems to be a reliable technique for demonstrating viral genome in the CSF. A few antiviral treatments have been proposed, however their efficacy is difficult to assess due to the low prevalence of the disease and the occurrence of rare cases with spontaneously prolonged survival.

Safety and efficacy of lamivudine-zidovudine combination therapy in antiretroviral-naive patients. A randomized controlled comparison with zidovudine monotherapy. Lamivudine European HIV Working Group.

OBJECTIVE: To compare safety and efficacy of lamivudine-zidovudine combination therapy with zidovudine monotherapy in treating human immunodeficiency virus type 1 (HIV-1)-infected, antiretroviral therapy-naive patients. DESIGN: Double-blind, randomized, multicenter, comparative trial of 129 patients throughout 24 weeks followed by 24 weeks of open-label lamivudine in combination with zidovudine. SETTING: Outpatients from 14 hospitals in Belgium, France, Germany, Spain, and the United Kingdom were enrolled within 6 months. PATIENTS: HIV-1-positive, antiretroviral-naive ( < or = 4 weeks prior zidovudine use) patients aged atleast 18 years with CD4+ cell counts between 0.10 x 10(9)/L and 0.40 x 10(9)/L (100-400/microL). INTERVENTION: Patients received either 300 mg of lamivudine every 12 hours in combination with 200 mg of zidovudine every 8 hours for 24 weeks or zidovudine monotherapy for 24 weeks. All patients were then allowed to receive zidovudine in combination with open-label lamivudine (300 mg every 12 hours). MAIN OUTCOME MEASURES: Efficacy was assessed by changes in CD4+ cell counts beta 2-microglobulin, neopterin, HIV-1 immune-complex dissociated (ICD) p24 antigenemia, and HIV-1 viral load. Safety was assessed by incidence of adverse clinical events and defined laboratory-measured toxic effects. RESULTS: Combination therapy showed superior treatment effects compared with monotherapy during the first 24 weeks as documented by changes in CD4+ cell counts (increase of 0.08 x 10(9)/L vs 0.02 x 10(9)/L; P < .001), ICDp24 (-88% vs -49%; P = .04), cellular viremia (-1.27 vs -0.20 log10 median tissue-culture infected dose [TCID50] per 10(6) peripheral blood mononuclear cells; P = .001), and viral load measured by HIV-1 RNA polymerase chain reaction using a Roche method (-1.33 vs -0.57 log10 copies/mL; P = .001) or an immune-capture method (-0.6 vs -0.14log10 copies/mL; P = .008). Observed changes were sustained to 48 weeks for patients continuing to receive combination therapy. Patients switching to receive combination therapy at week 24 showed improvements in CD4+ cell count and viral load to week 48. Mutation results suggested that mutations associated with zidovudine resistance may have developed more slowly over the first 24 weeks in patients receiving combination therapy. In contrast, mutations associated with lamivudine resistance appeared to develop rapidly, despite sustained antiviral treatment effect. However, the number of patients evaluated for genotypic changes was small, and confirmation of these results is needed in larger studies. No statistically significant differences in incidence or severity of clinically manifested or laboratory-measured toxic effects were noted between treatment groups. CONCLUSIONS: The combination of lamivudine and zidovudine results in a potent and sustained antiviral effect in antiretroviral-naive patients that is superior to that observed with zidovudine monotherapy.

Phase I/II study of 3TC (lamivudine) in HIV-positive, asymptomatic or mild AIDS-related complex patients: sustained reduction in viral markers. The Lamivudine European HIV Working Group.

OBJECTIVE: To evaluate the efficacy of 3TC (lamivudine), a synthetic nucleoside analogue that inhibits HIV reverse transcriptase in vitro, as treatment for HIV-positive, asymptomatic or mild AIDS-related complex patients. DESIGN: Open-label, multinational and multicentre, non-comparative, escalating dose study. METHODS: Patients who meet the selection criteria (n = 104) were enrolled in three European countries. Ten to 15 patients were included at each of the six dose levels of 3TC (0.5, 1.0, 2.0, 4.0, 8.0, 12.0 and 20.0 mg/kg daily in two divided doses every 12 h). Virological parameters--immune-complex dissociation (ICD) assay for HIV p24 antigenaemia, plasma HIV RNA load, whole blood assay and cellular viraemia--were evaluated at weeks 0, 4, 12 and 24. RESULTS: Sustained reductions in HIV RNA load and in ICD p24 antigen levels were observed and maintained over the 12-week assessment period. Greater reductions were noted at higher doses but this trend did not reach statistical significance. In 38 patients, reductions of cell viraemia were significantly greater at 4 weeks for patients treated at higher doses of 3TC. CONCLUSION: These virological data show that 3TC is a potent inhibitor of HIV replication in HIV-positive, asymptomatic or mild ARC patients as assessed by ICD p24 antigenaemia, plasma HIV RNA load and cell viraemia.

Photodynamic inactivation of cell-free HIV strains by a red-absorbing chlorin-type photosensitizer.

We have investigated the photodynamic activity of a new chlorin-type photosensitizer on a reference human immunodeficiency virus type 1 (HIV-1) strain, two wild-type HIV-1 isolates and two drug-resistant HIV-1 isolates. This chlorin was highly effective for the inactivation of free viruses, as assessed by two different quantitative cell culture assays. In the absence of blood components, all the HIV strains, including wild-type and drug-resistant mutant isolates, were totally inactivated using 30 micrograms ml-1 of chlorin and 0.75 J cm-2 of 661 nm light. Successful killing of HIV-1 strains in either plasma or whole blood was also obtained by increasing the chlorin concentration moderately. Our results demonstrate the antiviral efficiency of this chlorin, suggesting the potential application of dye-sensitized photoirradiation to decontaminate blood products.

Evaluation of safety and efficacy of 3TC (lamivudine) in patients with asymptomatic or mildly symptomatic human immunodeficiency virus infection: a phase I/II study.

In a phase I/II study, 7 levels of 3TC therapy (from 0.5 to 20.0 mg/kg/day) were studied in 104 asymptomatic and mildly symptomatic human immunodeficiency virus-infected patients with CD4 cell counts < or = 400 x 10(6)/L. Mild and transient episodes of diarrhea, headache, fatigue, nausea, and abdominal pain were the most frequent events reported. No dose-limiting toxicities were observed. Small and transient increases in CD4 cell counts were detected during the first 4 weeks of treatment. These were followed by progressive declines during prolonged therapy. Sustained decreases in beta 2-microglobulin, neopterin, and p24 antigen levels were seen over the 52-week study. There was no consistent dose-response correlation for any surrogate marker. Penetration of 3TC into cerebrospinal fluid (CSF) was in the same range as reported for ddC and ddI; the mean CSF-to-serum ratio was 0.06. These findings indicate that 3TC exhibits an excellent safety profile and has antiretroviral activity at the dosages studied.

HIV-1/HIV-2 seronegativity in HIV-1 subtype O infected patients.

Nine patients with atypical HIV-1 western blot profiles were diagnosed as having HIV-1 subtype O infection. All the patients were living in France; eight originated from Cameroon and one from France. Lymphocyte DNA amplification by the polymerase chain reaction was only positive when HIV-1 subtype O specific primers were used. Preliminary sequence analysis of amplified products and serological reactivity against a specific subtype O synthetic env peptide confirmed HIV-1 subtype O infection. HIV-1/HIV-2 enzyme-linked immunosorbent assays, especially those based on env peptides or on the sandwich format, can be negative in HIV-1 subtype O infection.