Simultaneous progressive multifocal leukoencephalopathy, Epstein-Barr virus (EBV) latent infection and cerebral parenchymal infiltration during chronic lymphocytic leukemia.

We report a non-HIV patient who had B chronic lymphocytic leukemia (CLL) with progressive multifocal leukoencephalopathy (PML) and diffuse cerebral leukemic parenchymal infiltration in the presence of JC virus and Epstein-Barr virus (EBV) cerebral co-infection. Multiple subcortical hypodensities lining the cortico-subcortical junction were present within the white matter on computerized tomography (CT) scan, with large areas of high signal intensity on T2-weighted sequences on magnetic resonance imaging (MRI). JCV DNA was identified in peripheral blood nuclear cells and cerebrospinal fluid polymerase chain reaction (PCR) DNA/DNA hybridization plus Southern blot analysis. Frontal stereotactic biopsy confirmed the diagnosis of PML by immunocytochemistry, in situ hybridization (ISH) with JC Enzo probe and electron microscopy. Leukemic B cells with the same phenotype as leukemic blood cells were disseminated in the demyelinated areas. They were labeled by anti-latent membrane protein and by BamHl W EBV probe after ISH. Adhesion and activation molecules were positive for CD23. Autopsy showed diffuse visceral leukemic infiltration without acutization. EBV-transformed B lymphocytes would favour JCV penetration and/or intracerebral reactivation of previously latent JCV infection with further development of simultaneous PML and cerebral CLL infiltration in an immunosuppressed patient.

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.

Susceptibility of HIV-1 isolates to zidovudine: correlation between widely applicable culture test and PCR analysis.

Thirteen isolates of human immunodeficiency virus type 1 (HIV-1) obtained in coculture with peripheral blood lymphocytes were tested for in vitro susceptibility to zidovudine (ZDV). Seven isolates were obtained from patients who had never been treated with ZDV and six from patients receiving the drug. The seven isolates from untreated patients and four of six from treated patients were susceptible to ZDV. The two isolates from the patients treated for the longest periods were resistant to the drug. The presence of mutations at critical positions of the reverse transcriptase gene was investigated by direct sequencing of polymerase chain reaction (PCR)-amplified DNA and four isolates were found to be mutants. An isolate from an untreated patient showed a change at residue 70 of the reverse transcriptase and an isolate from a patient treated for 4 months showed a change at residue 67. A change at residue 215 was found only for the two drug-resistant isolates, which correlated with the results obtained by Larder et al. using isolates from MT-2 cell cocultures. These results suggest that any HIV isolate provided by conventional coculture could be confidently tested for ZDV susceptibility in order to study the emergence of resistance during long-term therapy.

HIV-1 sensitivity to zidovudine: a consensus culture technique validated by genotypic analysis of the reverse transcriptase.

In order to select and standardize a reliable assay for the analysis of sensitivity of HIV isolates to AZT, we have compared two culture methods. The first assay (Cell-Associated Isolate Sensitivity Assay: CAISA) quantified AZT-resistant HIV isolates by end-point dilution cultures of peripheral blood mononuclear cells (PBMCs) in the presence of various concentrations of AZT. In the second assay (Cell-Free Isolate Sensitivity Assay: CFISA), following a conventional isolation of HIV, dilutions of infected cell-free supernatants were cultivated with fresh normal donor PBMCs in the presence of increasing concentrations of AZT. Samples from 64 untreated and AZT-treated patients were studied by CAISA (41), CFISA (43) or both assays (20). The CFISA, which allows the determination of titration parameters with respect to various kinetics patterns of viral replication was selected, and some of the CFISA phenotypically characterized isolates were further studied by nucleotide sequence analysis of the reverse transcriptase gene. CFISA showed that isolates from untreated patients were susceptible to AZT while the frequency of resistance increased with the duration of therapy. Genotypic analysis of CFISA-resistant isolates exhibited mutations at crucial positions, particularly at residue 215. We consider CFISA as a consensus culture technique for longitudinal studies of isolates from patients receiving AZT or other analogs of nucleosides.

Perspectives in antiviral chemotherapy.

The current progress in antiviral therapy is related to our better understanding of the viral multiplication, with potential targets for specific antiviral action at each step of the multiplication cycle inside the infected cell. Amantadine and Rimantadine are anti-influenza A drugs interfering with the penetration and the release of the virus. Most of the other antiviral drugs which are clinically available have the same target in common, namely the viral DNA polymerase. This holds true for modified nucleosides such as Acycloguanosine (Acyclovir), DHPG, Adenine-Arabinoside, Azidothymidine as well as pyrophosphate derivatives such as phosphonoformic acid. Unfortunately the antiviral chemotherapy must confront 3 obstacles: 1) a possible interference with the normal cellular metabolism, leading to residual cytotoxic side effects; 2) the genetic variability of the viruses, producing drug-resistant mutants and 3) the inability of any antiviral chemotherapeutic agent known to date to eradicate latent viral infection. A new approach of the control of latent infection is suggested with anti sense oligonucleotides of hybridons.

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.

Eczema herpeticum of the child. An unusual manifestation of herpes simplex virus infection.

Two children aged 7 months with eczema herpeticum received treatment consisting of intravenous acyclovir and human plasma with a high titer of herpes simplex virus antibodies. One recovered following two recurrences, but the other died rapidly, suffering both septicemia due to Pseudomonas aeruginosa and herpetic encephalitis. In both cases, the virus involved was a herpes simplex virus type 1 (HSV 1). The various isolates obtained before, during and after treatment remained equally sensitive to acyclovir. These observations highlight three points: the unpredictable and sometimes dramatic development of eczema herpeticum in the young child; the urgency of early diagnosis and treatment; the role of environment in viral contamination.

[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.

[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.

[Prospects and current data in antiviral chemotherapy]. / Perspectives et données présentes en chimiothérapie antivirale.

The advances achieved in our knowledge on the virus multiplication cycle and the challenge created by the advent of AIDS have resulted in a rational development of new antiviral agents. However, antiviral chemotherapy is hindered by several obstacles: (1) most antiviral drugs are cytotoxic, with the exception of acyclovir which owes its remarkable safety to its activation by the thymidine kinase of the herpes simplex viruses and of the varicella-zoster virus; (2) the risk of selecting resistant strains by mutation of viral enzymes is the unavoidable price to pay for the specificity of new antiviral agents which interfere with these enzymes; so far, this risk seems to apply only to immunocompromised patients; (3) latent viral infection, defined as the lack of active multiplication of the virus, cannot be eradicated by the antiviral drugs available at present since these drugs are inhibitors of viral multiplication. However, anti-sens oligonucleotides could, at least in theory, be used to treat this type of infection which is not limited to the Retroviridae or Herpesviridae families. Finally, antiviral drugs active against life-threatening infections that are as common as rabies and the severe forms of measles or viral hepatitis remain to be discovered.

[Virologic diagnosis of ocular infections]. / Diagnostic virologique des infections oculaires.

Viral infections of the anterior part of the eye are easy to diagnose as a rule: the inoculation of cell cultures with specimens collected on readily accessible and virus-rich lesions facilitates the isolation of adenoviridae, picornaviridae and herpesviridae responsible for these lesions. Viral infections of the posterior part of the eye are a different matter, and they require more complex and also more invasive investigation techniques. Yet an aetiological research is necessary: since effective antiviral agents with a highly specific mode of action are available, the virus to be eradicated must be identified with great accuracy.

[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.