Éditorial

Le numéro 12 de la revue Projets de paysages est constitué d’un dossier thématique consacré à l’interface paysage et culture, d’un dossier « Chercheurs en devenir » qui rassemble des articles issus des Journées doctorales du paysage (École de la nature et du paysage de Blois, octobre 2014), faisant état de problématiques de jeunes chercheurs ainsi que d’un « Varia ». Le dossier thématique « Paysage et culture » rassemble plusieurs articles portant sur la France, l’Amérique du Nord (les États-..

Etude de l'immunité liée au virus de la rougeole au moyen de l'hémagglutinine recombinante et de protéines chimériques à épitopes multiples

Doctorat en sciences médicalesinfo:eu-repo/semantics/nonPublishe

A simplified immunoassay based on measles virus recombinant hemagglutinin protein for testing the immune status of vaccinees

Simplified tests based on recombinant antigens are considered to be important for monitoring immunity against measles virus (MV). The hemagglutinin protein (H) is the main target for neutralising and protective antibodies. We produced a recombinant MV-H protein, in a high-yield mammalian expression system based on the Semliki Forest virus replicon. The antigenicity of this recombinant protein was investigated with monoclonal antibodies and its suitability for measuring the immune status of vaccinees was tested in a large cohort by ELISA (H-ELISA). The results were evaluated against neutralisation (NT) and hemagglutination inhibition (HI) titers and MV-specific IgG measured in a commercial whole-virus based ELISA (MV-ELISA, Enzygnost). The H-ELISA correlated better with HI (r=0.78) and NT titers (r=0.80), than the MV-ELISA (HI, r=0.58; NT, r=0.59). In contrast to the MV-ELISA, the H-ELISA detected no false-positive sera (P<0.02) and the number of false-negative sera was significantly lower in the H-ELISA than in the MV-ELISA (4/378 vs. 15/378; P<0.025). The performance of the H-ELISA did not deteriorate significantly when, instead of background corrected net values, uncorrected raw O.D. values of the H-antigen were considered, or when early time points (30 min) were evaluated. These results demonstrate that the recombinant H-ELISA detects efficiently non-immune individuals among vaccinees, despite their relatively low MV-antibody levels. A simplified format with single value measurements did not result in loss of sensitivity or specificity and its performance compared favorably with commercial ELISAs based on whole virus. Copyright (C) 1998 Elsevier Science B.V.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Modulation of the metabolism and adverse effects of benzo[a]pyrene by a specific antibody: A novel host factor in environmental carcinogenesis?

The influence of specific antibodies on molecular and cellular mechanisms of activation, detoxification and biological activity of the ubiquitous carcinogen benzo[a]pyrene (B[a]P) was investigated using a monoclonal antibody. The antibody was shown to decrease cellular uptake and metabolic activation of B[a]P as demonstrated by higher recovery of unmetabolized B[a]P and decreased formation of end-point phenol metabolites in two types of target cells. Furthermore, strong antibody reactivity with 7,8-diol-B[a]P provided a second chance for interrupting metabolic activation by sequestration of this intermediate metabolite in the extracellular space. The biological relevance of B[a]P and 7,8-diol-B[a]P redistribution by antibody was demonstrated by reversion of B[a]P-induced inhibition of proliferation of human peripheral blood lymphocytes and by inhibition of CYP 1A1 induction in HepG2 cells. Remarkably, the antibody was still protective against B[a]P-induced immunotoxicity even after delayed addition, suggesting a more important role of metabolites in immunotoxicity than has been appreciated so far. Although B[a]P is activated to 7,8-diol-B[a]P in the same cells that are inhibited by this metabolite, the antibody completely restored lymphocyte proliferation indicating that extracellular trapping of the 7,8-diol-B[a]P is biologically highly effective. Thus, repartitioning of both B[a]P and its metabolites by the antibody may reduce their effective concentration in susceptible target organs and therefore relieve overloaded DNA repair mechanisms and inhibit carcinogen-induced P450 induction. These in vitro data also suggest that a natural or prophylactic antibody response against carcinogens may be associated with a reduced risk of cancer. © Oxford University Press 2005; all rights reserved.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Immunosorbent Assay Based on Recombinant Hemagglutinin Protein Produced in a High-Efficiency Mammalian Expression System for Surveillance of Measles Immunity

Recombinant hemagglutinin (H) protein of the measles virus (MV) was produced in mammalian cells with a high-yield expression system based on the Semliki Forest virus replicon. Crude membrane preparations of H protein-transfected BHK-21 cells were used to coat microtiter plates to measure specific immunoglobulin G antibodies in 228 serologically defined serum samples mainly from measles late-convalescent adults. The titers by the enzyme-linked immunosorbent assay for the H protein (H-ELISA) closely correlated with neutralization test (NT) titers (R(2) = 0.66), hemagglutination inhibition test (HI) titers (R(2) = 0.64), with the titers from a certified commercial ELISA based on whole MV-infected cells (MV-ELISA; R(2) = 0.45). The correlations described above were better than those of the commercial MV-ELISA titers with the NT (R(2) = 0.52) or HI (R(2) = 0.48) titers. By using the 2nd International Standard for anti-measles serum, the detection level of the assay corresponds to 215 mIU/ml for undiluted serum, which corresponds to the estimated threshold for protective immunity. The specificity, accuracy, and positive predictive value were, in general, better for the H-ELISA than for a commercial MV-ELISA, independent of whether HI, NT, or HI and NT were used as “gold standards.” In contrast, the H-ELISA proved to be slightly less sensitive than the MV-ELISA (sensitivities, 98.6 versus 99.5%, respectively; P was not significant). The assays did not differ significantly in the number of serum samples with positive HI and NT results (n = 212) which measured false negative (H-ELISA, 2 of 212 [0.94%]; MV-ELISA, 1 of 212 [0.47%]), but the H-ELISA detected significantly more measles-susceptible individuals than the MV-ELISA (10 of 11 versus 3 of 11, respectively; P < 0.05) among the individuals whose sera had negative HI and NT results. Our data demonstrate that the H-protein preparation that we describe could be a cost-effective alternative to current whole-virus-based ELISAs for surveillance for immunity to measles and that such an assay could be more efficient in detecting susceptibility to measles. Furthermore, unlike whole MV-based antigens, H-protein would also be suitable for use in the development of a simple field test for the diagnosis of measles

Evaluation of Hemagglutinin Protein-Specific Immunoglobulin M for Diagnosis of Measles by an Enzyme-Linked Immunosorbent Assay Based on Recombinant Protein Produced in a High-Efficiency Mammalian Expression System

Recombinant hemagglutinin (H) of the measles virus (MV) expressed in a mammalian high-expression system based on the Semliki Forest virus replicon was used in an enzyme-linked immunosorbent assay (ELISA) for the detection of specific immunoglobulin M (IgM) and IgG in patients with acute-phase measles. One hundred twelve serum specimens from 70 patients with measles were analyzed. Case definition was based on a commercial IgM ELISA that utilizes MV-infected cells (MV-ELISA) (Enzygnost; Behring Diagnostics); the clinical criteria of the Centers for Disease Control and Prevention (Atlanta, Ga.); and/or the increase in hemagglutinin test titers, neutralization test titers, and levels of MV-specific IgG whenever paired sera were available. The initial time courses of the IgM signal after the onset of rash are similar in the H- and MV-ELISAs. On days 0 to 19, both ELISAs detected IgM in 67 of 68 (98.5%) sera. Average maximal levels of IgM seem to persist, however, about 10 days longer in the MV-ELISA (up to day 25) than in the H-ELISA (day 15). From days 20 to 29 and 30 to 59, the H-ELISA detected only 64.3 (9 of 14) and 19.2% (5 of 26), respectively, of sera that were IgM positive by MV-ELISA. At least up to day 30, the performance of the H-ELISA seemed to be similar to that reported for commercial ELISAs based on whole MV. Our results demonstrate that MV H-specific IgM can be used to diagnose most measles cases from a single serum specimen collected within 19 days after the onset of rash and that the recombinant protein used in this study is suitable for this purpose

Hepatic regenerating nodules: A mimic of recurrent cancer in children

Background. Pseudometastatic lesions of the liver may be discovered incidentally in children previously treated for malignant tumour. Objective. To describe the radiological pattern of these lesions and to analyse their pathogenesis. Materials and methods. Nine children, 2-12 years' old at the time of diagnosis, are described in this retrospective multicentre report. The primary tumours were: nephroblastoma (n = 2), neuroblastoma (n = 2), Ewing's tumour/PNET (n = 2), non-Hodgkin's lymphoma (n = 1), and osteosarcoma (n = 2), treated by surgery (8/9), chemotherapy (9/9), intensive chemotherapy and bone-marrow transplantation (5/9), and radiotherapy (7/9). Three children suffered veno-occlusive disease (VOD) during treatment. The hepatic assessment was performed by sonography (8/9), Doppler (7/9), multiphase spiral CT (8/9) and MRI (7/9). Results. Lesions were discovered 15 months to 16 years after completing treatment. CT was the most sensitive modality for diagnosis. Lesions were multiple in eight cases, measured 2-50 mm, and appeared hypervascular on the arterial phase of CT and/or MRI in every case. Metastases were excluded on the basis of histological verification (n = 2) and clinical and radiological follow-up. Conclusion. Pseudometastatic hypervascular hepatic nodules can appear after treatment of a malignant tumour in children. The hypothesis of benign regenerative lesions secondary to treatment and/or VOD is considered.SCOPUS: ar.jinfo:eu-repo/semantics/publishe