[Tropism of hepatitis C virus for leukocytes-- importance of the analysis of viral E1 and E2 envelope glycoprotein genes by sequencing]. / Tropisme leucocytaire du virus de l'hépatite C - intérêt de l'analyse des séquences des gènes des glycoprotéines d'enveloppe virales E1 et E2.

The ability of hepatitis C virus (HCV) to infect leukocytes could favour HCV pathogenesis. Although viral infection of these immunocompetent cells is poorly (or not) productive, the impact on their immunomodulatory functions could be important. Viral envelope glycoproteins E1 and E2, because of their crucial role in the recognition of viral receptors on permissive cells, could contribute to viral leukocytic tropism and, as a consequence, to the pathophysiology of HCV chronic infection.

Atypical symptoms in patients with herpesvirus DNA detected by PCR in cerebrospinal fluid.

BACKGROUND: Polymerase chain reaction (PCR) detection of herpesvirus DNA in cerebrospinal fluid (CSF) is an important tool in the diagnosis of central nervous system (CNS) syndromes. The corresponding viral infections present with diverse clinical signs, which are often classical although no sign can be considered as specific. This retrospective study aims to describe atypical symptoms in patients with herpesvirus DNA detected in CSF by PCR. A total of 3452 cerebrospinal fluid samples from patients with suspected herpesvirus infection of the CNS were investigated between 1998 and 2003 in our clinical virology laboratory. "In-house" PCRs for each herpesvirus [herpes simplex virus (HSV), varicella zoster virus (VZV), cytomegalovirus (CMV), Epstein Barr virus (EBV), or human herpes virus 6 (HHV6)] were used until 2001 and a commercially available "Herpes Consensus PCR" was used thereafter. One of the five herpesviruses investigated in this study was found in 71 (2.1%) of CSF samples (37 HSV, 14 VZV, 1 CMV, 9 EBV and 10 HHV6). These samples were obtained from 62 patients whose clinical findings were generally consistent with the PCR data. However, some little known features of herpesvirus-related symptoms, such as partial seizure associated with HSV infection, and unusual VZV or HHV6-related myelitis were also observed.

[Rational use of viral serology in the child]. / Utilisation rationnelle des sérologies virales chez l'enfant.

Very sensitive and allowing discrimination between IgG and IgM, the enzyme immunoassays are the most commonly used to carry out viral serologies. The presence of IgG corresponding to a natural or postvaccination immune status or to a latent or occult infection is a reliable information to assess the protection against some opportunistic or teratogenic infections respectively in children or pregnant women. The search of IgM by immunocapture is suitable to early diagnose the majority of common viral diseases in childhood, specially when presenting in an atypical form, and to attest a congenital rubella infection. Moreover, IgG avidity proved to be useful in dating an infection and confirming positive IgM results. However, viral infections localized at an entry site of the host, such as respiratory and enteric diseases, as well as syndromes of herpes simplex and enteroviral origin, do not represent indications of serological diagnosis. Provided its use with discernment, the serological diagnosis of viral infections have to be considered as a high performance and of low cost biological tool.

Evidence of feline immunodeficiency virus replication in cultured Kupffer cells.

OBJECTIVE: To determine if cultured feline Kupffer cells (KC) are as permissive for feline immunodeficiency virus (FIV) as cultured human liver macrophages are for HIV. Two types of infection likely to be relevant to the in vivo situation were used. KC were infected with either free virus or autologous infected peripheral blood mononuclear cells (PBMC). METHODS: Feline KC were isolated by centrifugal elutriation from collagenase-perfused liver; cultured cells were characterized by their morphological appearance and their erythrophagocytotic properties. After infection, viral replication was measured by enzyme-linked immunosorbent assay, reverse transcriptase activity, immunofluorescence assay, in situ hybridization and electron microscopic observations. RESULTS: Three days after isolation, 85% of cultured KC were able to internalize red blood cells; 45% were CD4-positive and 65% expressed a 24 kD protein thought to be a receptor for FIV (CD9). After the addition of autologous infected PBMC or cell-free supernatant of chronically infected IRC4 cells to KC cultures, a peak of viral replication was detected at day 28. Antigen revealed by immunofluorescence assay was present in only 0.4%, and viral RNA was detected by in situ hybridization in 2% of the infected cells. CONCLUSIONS: FIV can replicate in cultured feline KC without inducing any cytopathic effect, which suggests that these cells may play a role in the physiopathology of FIV infection.

Protection of SIVmac-infected macaque monkeys against superinfection by a simian immunodeficiency virus expressing envelope glycoproteins of HIV type 1.

The infection of macaque monkeys by attenuated simian immunodeficiency virus can vaccinate against pathogenic molecular clones and isolates of the same virus. The correlates of this potent protective immunity are not fully understood but may be the key to an effective AIDS vaccine for humans. Aiming to determine whether host immune responses to envelope glycoprotein are an essential component of the immunity to primate lentiviruses, we have tried to superinfect SIVmac-infected macaque monkeys with SHIVsbg, a chimeric primate lentivirus constructed from the SIVmac239 genome with the env, rev, tat, and vpu genes from HIV-1 Lai. After inoculation of a large dose of SHIVsbg, the chimeric virus was isolated by coculture of mononuclear blood cells from four of five SIV-infected monkeys, but three animals were protected from extracellular SHIV viremia and did not seroconvert to HIV-1 glycoproteins. In the two SIV-infected monkeys that did develop SHIV viremia, cell-associated viral load was reduced at least 100-fold. These data indicate that an antiviral response capable of effectively controlling primate lentivirus replication might not necessarily involve the envelope glycoprotein.

Enterovirus in sudden unexpected deaths in infants.

BACKGROUND: Conventional approaches to virus detection failed to provide convincing evidence of a viral etiology in sudden unexplained deaths in infants (SUDI). Many viruses may not have been detected by the routinely used methods; among them enteroviruses (EV) have seldom been found in SUDI. METHODS: In this study EV were sought directly in stools, in pharyngeal and tracheal samples and in myocardial and lung tissues, by using a nested PCR; they were also sought indirectly by detecting IgM antibodies with a new capture immunoassay. Twenty-four SUDI cases were divided into two groups: Group I, certainly associated with; or Group II, not associated with clinical, biologic or histologic signs of viral infection. RESULTS: EV were found in stools but their prevalence was not significantly different between Group I and Group II (20 and 22.2%, respectively). On the contrary EV were detected in respiratory tract and/or lung samples in 53.8% of infants of Group I and in none of Group II. Anti-EV IgM antibodies were detected in 55.5% of infants of Group I and in none of Group II. CONCLUSIONS: These results indicate that EV infection may be specifically associated with the subgroup of SUDI with viral signs, raising the question of its role in this condition.

Molecular epidemiology of ocular isolates of adenovirus 8 obtained over nine years.

Twenty nine strains of adenovirus 8 have been isolated over nine years in Strasbourg, France, 22 of which were from one private ophthalmologist. To assess a possible relation between these strains, the DNA of adenovirus was analysed by restriction fragment length polymorphism using eight different enzymes. Among these, three proved discriminant (Xba I, Bgl II, Eco RI) and made it possible to define 13 genotypes differing from each other by one to three DNA bands. Seven genotypes were unique isolates, while three, representing 16 strains, were isolated over five to eight years. All the genotypes but one were closely related, with 87% homology. All 13 differed from an adenovirus 8 strain from Lyon (homology 68-76%). This study confirmed the stability of adenovirus 8 in a given population.

Aspirin and heparin prevent hepatic blood stasis and thrombosis induced by the toxic glycoprotein Bolesatine in mice.

Bolesatine is a toxic glycoprotein isolated from Boletus satanas Lenz, which inhibits protein synthesis in vivo and in vitro. The LD50 (24 h) is 1 mg /kg bw (i.p.), in mice and rats. When given i.p. to mice (0.1 - 1.0 mg/kg bw) bolesatine induced thrombi and blood stasis in the liver, 5 - 21 h after injection, and modifications of the number of blood corpuscles in peripheral blood. These effects were efficiently reversed by aspirin, ticlopidin and heparin (as attested by histology and electron microscopy) which however failed to prevent death in animals given lethal doses. Together, these results showed that the death of bolesatine poisoned animals given high doses, was rather due to a combination of thrombosis and other toxic effects. In addition, they suggest that these antithrombotic drugs may overcome cases of human poisoning, with low exposures of this boletus, showing a hypertension probably due to mechanical obstruction which resists normal therapy.

[In vitro demonstration of a deficit in suppressor T-cell function in alcoholic cirrhosis. Role of hepatitis B virus infection]. / Mise en évidence in vitro d'un déficit de la fonction lymphocytaire T suppressive dans la cirrhose alcoolique. Rôle de l'infection par le virus de l'hépatite B.

The T lymphocyte suppressor cell activity has been evaluated in 33 alcoholic patients compared with 16 normal controls, using an in vitro test. Suppressor T cells were activated with concanavalin A, and suppressor effect was quantified by the inhibition of an autologous B cell culture response to Pokeweed Mitogen. When compared with controls, cirrhotic patients showed a significant defect of suppressor cell activity on B cell production of IgG (20 +/- 3 vs 46 +/- 5 p. 100, p less than 0.001) and IgM (26 +/- 4 vs 56 +/- 8 p. 100, p less than 0.05). In cirrhotic patients, defect of T cell suppressor function was independent of sex and severity of the cirrhosis (Child's staging). This defect was more marked in cirrhotics with serological markers of hepatitis B virus (HBV) infection (n = 11) than in cirrhotics without markers (n = 22) (9 +/- 5 vs 25 +/- 3 p. 100, p less than 0.05; 16 +/- 6 vs 30 +/- 5 p. 100, p less than 0.05 respectively for IgG and IgM production suppression). These results suggest that HBV and lymphocytes interact directly. This interaction could increase the T suppressor cell defect, and explain the promoting role of HBV infection in the constitution of the cirrhosis in alcoholics even when viral replication is not serologically apparent.

[Comparison of immunogenicity of vaccination and serovaccination against hepatitis B virus in patients with alcoholic cirrhosis]. / Comparaison de l'immunogénicité de la vaccination et de la sérovaccination contre le virus de l'hépatite B chez les malades atteints de cirrhose alcoolique.

OBJECTIVES: The association of anti-HBs immunoglobulins and anti-HBV vaccine could increase the immunogenicity of the latter. The aim of this prospective randomized trial was to compare the immunogenicity of anti-HBs vaccination and serovaccination in alcoholic patients with cirrhosis. METHODS: Alcoholic patients with cirrhosis were randomized in 2 groups: a) Vaccination group: 3 i.m. injections of GenHevac B followed by one booster at month 9; b) Serovaccination group: same vaccination schedule followed by one i.m. injection of anti-HBs immunoglobulins (500 IU). RESULTS: Twenty-five patients (17 males and 8 females, mean age 56 years) were included in the study: 13 received a vaccination and 12 received a serovaccination. After 12 months, the seroconversion rates were 69% and 67% in vaccination and in serovaccination groups, respectively. The predictive factors of non responsiveness were as following: Child B cirrhosis, low number of CD8, a high CD4/CD8 rate, the existence of HLA DR7 antigen, and the absence of HLA DR1 antigen. CONCLUSION: In alcoholic patients with cirrhosis, serovaccination does not increase the immunogenicity of anti-HBs vaccination and should not be recommended.

[Hepatitis B virus, serological markers of viral infections and humoral immunity in alcoholic cirrhosis]. / Virus de l'hépatite B, marqueurs sérologiques d'infections virales et immunité humorale au cours de la cirrhose alcoolique.

In order to define the role of hepatitis B virus (HBV) in alcoholic liver disease and to study the relationship between HBV and other common viruses, the serological markers of viral disease (HBV, Rubella, Polio, Herpes, and Cytomegalovirus-CMV) were compared in 163 patients with alcoholic cirrhosis (group C), 100 patients with alcoholic steatosis (group S) and in 168 non-alcoholic control subjects (group NA). A significantly increased prevalence of HBV markers in group C was related to the presence of anti-HBc antibodies, in 10.5 p. 100 of cirrhotic patients, vs. 1.2 p. 100 in group S and 1 p. 100 in group NA (p less than 0.01). In cirrhotic patients with HBV markers (HBV +) incidence of alcoholic hepatitis was 4 times lower and the total duration of alcohol overconsumption was significantly lower than in cirrhotic patients without these markers (HBV-). Hepatic function tests were not different in HBV + and HBV- cirrhotic patients, excepted for the ASAT/ALAT ratio (1.55 +/- 0.10 vs. 1.92 +/- 0.12; p less than 0.05). Prevalence of anti-CMV antibodies, and anti-herpes greater than 1/100 antibodies, was significantly increased in S and C groups (p less than 0.01). Anti-Rubella, Polio, and CMV antibody titers were higher (p less than 0.05) in HBV + than in HBV- cirrhotic patients. In cirrhotic subjects, titers of these 3 anti-virus antibodies were not related to alcoholic hepatitis or to IgG and IgM concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

[Determination of sub-populations of circulating T lymphocytes in alcoholic cirrhosis using monoclonal antibodies OKT3, 4, 5, Leu 2 and Leu 15. Effect of hepatitis B virus infection]. / Détermination des sous-populations lymphocytaires T circulantes au cours de la cirrhose alcoolique par les anticorps monoclonaux OKT3, 4, 8, Leu 2 et Leu 15. Influence de l'infection par le virus de l'hépatite B.

Peripheral T lymphocyte subpopulations were quantified in 24 alcoholic cirrhotic patients, 11 of them having anti-HBs and/or anti-HBc antibodies, and were compared with 35 healthy control subjects, 10 of them having anti-HBs and/or anti-HBc antibodies. The monoclonal antibodies utilized (OKT3, OKT4, OKT8 in simple staining, Leu 2 and Leu 15 in double staining) are considered as markers of mature (CD3), helper (CD4), cytotoxic/suppressor (CD8, Leu 2), suppressor (Leu [2+ 15+), and cytotoxic (Leu 2+ 15-) T cells. In cirrhotics, when compared to controls, the number of CD3 cells was reduced (p less than 0.01); the proportion of CD4 cells was within normal range, and that of CD8 cells diminished (p less than 0.001), contrasting with an increased proportion of Leu 2+ cells (p less than 0.01), related to an increased proportion of Leu 2+ 15+ cells. Leu 2+ 15- lymphocytes were within normal range. In control subjects, a decreased proportion of Leu 2+ 15+ cells was found (p less than 0.05) when Ac HBs and/or Ac HBc were present. In cirrhotics having at least one serologic marker of hepatitis B virus infection, when compared with negative ones, increased proportions of Leu 2+ (p less than 0.05) and Leu 2+ 15+ (p less than 0.05) cells were found. These results show that data concerning T lymphocyte subpopulations are conflicting when various types of antibodies are used. However, they suggest abnormalities of immune regulation, possibly a defect of T suppressor cell function. Hepatitis B virus infection probably modifies immune regulation in alcoholic cirrhosis, and perhaps in normal subjects.

Reduction of the observed prevalence of so-called non-A non-B hepatitis using sensitive markers of HBV and herpes viruses infections.

In the absence of a specific marker, the observed prevalence of so called non-A non-B hepatitis depends on the sensitivity of the markers of the other viral infections known to induce hepatitis. We have reevaluated this prevalence after using sensitive markers of HBV (HBs monoclonal radioimmunoassay M-RIA and IgM anti-HBc), EBV (IgM anti-VCA), CMV (IgM anti-CMV) and HSV (IgM anti-HSV) in a group of 53 subjects usually considered as having acute or chronic hepatitis. Detection of IgM against HBc, CMV and HSV used immunocapture tests. Among the 37 patients with acute hepatitis, 11 (30 p. 100) were positive for at least one sensitive marker, including 10 markers of HBV (7 M-RIA and 3 IgM anti-HBc) and one IgM anti-CMV. Among the 16 patients with chronic hepatitis, one was positive for HBV by M-RIA; five patients had a false positive reaction to EBV (IgM anti-VCA) disappearing when rheumatoid factor was eliminated. This study shows that many cases of the so-called non-A non-B hepatitis are in fact due to HBV or to a variant of HBV. Definition of non-A non-B hepatitis must include subjects negative for HBV by M-RIA and IgM anti-HBc and negative for CMV by IgM anti-CMV.

[Tick-borne encephalitis in Alsace]. / L'encéphalite à tiques en Alsace.

Central European tick-borne encephalitis is mainly found in Central European countries and Austria where hundreds of cases are reported each year. Apart from 2 cases diagnosed in Alsace in 1968 and 1970 respectively, this disease was hitherto unknown in France. We report 8 new cases observed in Alsace between 1985 and 1990. Clinical presentation in these 10 patients was a pure meningitis syndrome in 4 cases and meningo-encephalitis in 6 cases, very severe in 3 of them. All patients recovered rapidly, and only 3 have slight sequelae. In a seroprevalence survey conducted in 1989 among 619 professional foresters of Eastern France, 8% were found to be seropositive, which suggests that the disease is often unrecognized. A study of the large series published in Austria and in other Central European countries has shown that the prognosis of tick-borne encephalitis is not always as favourable as it was in the Alsatian cases: severe sequelae or death occur in 1 to 2% of the patients. The need for a better detection of the disease and for vaccination of the subjects at risk must be emphasized.

[Isolation of 503 strains of virus in meningeal syndromes. Virologic and epidemiologic study]. / Isolement de cinq cent trois souches de virus au cours de syndromes méningés. Etude virologique et épidémiologique.

An epidemiological and virological study on meningitis in Alsace during the 13-year period 1968-1981 is reported. 503 viruses isolated were associated with the diagnosis of meningitis. The Echovirus type 30 accounted for 48 p. 100 of the cases, the mumps virus of 12 p. 100. The remarkable high incidence of Echo 30 virus-associated cases is related to three outbreaks which occurred in Alsace during the summers of 1968, 1975 and 1980. Children below 12 years of age are the most sensitive population and particularly boys who made up 2/3 of the number of cases.

[Confusional form of Chlamydia psittaci encephalitis. Diagnostic value of microimmunofluorescence. A case]. / Forme confusionnelle d'une encéphalite à Chlamydia psittaci. Intérêt diagnostique de la micro-immunofluorescence. Une observation.

A rare case of Chlamydia psittaci encephalitis is reported. The disease started with anxiety, agitation and fever (38.5 degrees C) accompanied with hallucinations and regressed within 48 hours, but a confusional syndrome persisted for 9 days. Alterations in the blood-brain barrier with low CSF protein levels and signs of lateralization could be demonstrated. The inflammatory syndrome remained discrete. The cause of the disease was disclosed by serology and epidemiological investigations. The spontaneous outcome was favourable, thus confirming that neurological forms of psittacosis are benign. This case shows that microimmunofluorescence is more sensitive than the complement fixation test and that significant levels of antibodies directed against Chlamydia psittaci may persist for almost one year in the absence of treatment.

[Meningoencephalitis in a primary cytomegalovirus infection. Localization to the brain stem with bilateral subclinical optic neuritis]. / Méningo-encéphalite lors d'une primo-infection due au cytomégalovirus. Localisation au tronc cérébral avec névrite optique infraclinique bilatérale.

A female adult without previous medical or surgical pathology nor specific treatment developed meningoencephalitis localised to the brainstem associated with bilateral subclinical optic neuritis. Serology showed that the disease was related to primary infection with cytomegalovirus. The patient spontaneously recovered.