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Biomedical subjects

E Thiry

Publications and source records attributed to E Thiry.

At least 73 records · Page 4Linked to original sources

Epidemiological evaluation of a monoclonal ELISA detecting antibodies against bovine leukaemia virus in serum pools.

Sensitivity, specificity and predictive values of an enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against bovine leukaemia virus (BLV) were evaluated using a representative sample of 145 serum pools, comprising from 3 to 48 individual sera. The sample was constituted according to the frequency distribution of the negative and positive pools analysed during a screening involving the whole cattle population of Belgium. Sensitivity and specificity were estimated to 88.9% and 100% and the predicted negative and positive values were 99.9% and 100%, respectively. These results indicate the use of serum pools is suitable for the detection of BLV infected herds in eradication campaigns.

Animals↗

Attachment of the gammaherpesvirus bovine herpesvirus 4 is mediated by the interaction of gp8 glycoprotein with heparinlike moieties on the cell surface.

Cell surface heparan sulfate serves as the initial receptor for several alphaherpesviruses and at least one betaherpesvirus. This study shows that during the process of adsorption of the gammaherpesvirus bovine herpesvirus 4 (BHV-4), the viral glycoprotein gp8 interacts with heparinlike moieties of cell surface. This conclusion is based on the following findings. (i) Soluble heparin was capable of blocking BHV-4 infection of Georgia bovine kidney cells by inhibition of viral attachment. (ii) Nevertheless, after virus adsorption to Georgia bovine kidney cells, heparin was partially capable of removing adsorbed virus. (iii) Enzymatic digestion of cell surface heparan sulfate but not of chondroitin sulfates A, B, and C reduced the binding of the virus to the cells, and rendered the cells partially resistant to infection. (iv) Radiolabeled purified BHV-4 bound to wild-type Chinese hamster ovary cells, whereas binding of the virus to mutant Chinese hamster ovary cell lines that where deficient in either all glycosaminoglycans or only heparan sulfate was significantly impaired. (v) Using heparin-affinity chromatography, gp8 glycoprotein was shown to bind specifically to immobilized heparin and to elute in the presence of soluble heparin. These data together showed that the gammaherpesvirus BHV-4, like alphaherpesviruses and one betaherpesvirus, adsorbs to cells by binding to cell surface heparin-like moieties. Therefore, this study extends the group of herpesviruses interacting with heparinlike moieties at the cell surface to a member of the gammaherpesvirinae subfamily.

Adsorption↗

Identification of 108K, 93K, and 42K glycoproteins of bovine herpesvirus-1 by monoclonal antibodies.

Three glycoproteins of bovine herpesvirus-1 (BHV-1) other than glycoproteins gI, gIII, and gIV were identified by monoclonal antibody (MAb) analyses. Monoclonal antibodies were obtained by immunization of mice with either BHV-1 envelope or virus infected cells, from which the glycoproteins gI, gIII, and gIV were removed by immunoaffinity. In the latter immunization procedure mice were tolerized either against normal cellular antigens with or without glycoproteins gI, gIII, gIV, and nucleocapsid. From 154 anti-BHV-1 hybridomas isolated, 39 MAbs precipitated a 108K glycoprotein. Two other glycoproteins of respectively 42K and 93K were precipitated each by one MAb. These three glycoproteins were detected in infected cell lysate. Nine anti-108K glycoprotein MAbs neutralized BHV-1 infectivity and three non-neutralizing MAbs were able to reduce plaque development when virus was grown in the presence of these MAbs. It is therefore suggested that this glycoprotein is involved in viral entry into the cell and in cell-to-cell spread of the virus.

Animals↗

Personal medical and social data: their processing and legal protection.

Computerized tools are, just like other machines, very useful to man. However the development of data processing requires regulation especially oriented towards the principles of the protection of private life and of the guarantees attached thereto. An analysis of the international legal tools relating to the protection of human rights confirms the constant concern for individual rights and the interests of society. Belgian legislation relating to the computerized processing of medical and social personal data, in some respects, reveals the dangers of a lack of protection of the individual's private life. In conclusion, this article points out that in a modern state one should remain vigilant because any blow to human rights is a perversion leading to the decay of law.

Belgium↗

Identification of different target glycoproteins for bovine herpes virus type 1-specific cytotoxic T lymphocytes depending on the method of in vitro stimulation.

Vaccinia virus recombinants expressing the three major bovine herpes virus-1 (BHV-1) glycoproteins gI, gIII and gIV were used to identify the major target antigens for BHV-1-specific CTL isolated from immune cattle. Peripheral blood mononuclear cells (PBMC) expanded in vitro in the presence of interleukin-2 (IL-2) and lysed both gIII- and gIV-infected target cells. Secondary in vitro stimulation of PBMC was also performed in the presence of either fixed BHV-1-infected autologous fibroblasts or ultraviolet (UV)-inactivated virus. Both methods of antigen presentation allowed the proliferation of BHV-1-specific CTL but the target glycoprotein for these CTL differed depending on the method of stimulation. Vaccinia-gIV-infected targets were lysed predominantly when PBMC were stimulated by fixed infected fibroblasts, whilst PBMC stimulated by UV-inactivated virus lysed mostly vaccinia-gIII-infected targets. This observation could be explained by a different processing pathway of BHV-1 antigens in each cell type involved.

Animals↗

Effect of repeated intradermal injections of bovine herpesvirus type 1 antigen on seronegative cattle.

Forty-three cattle seronegative to bovine herpesvirus-1 (BHV-1) were given from one to five intradermal injections of BHV-1 inactivated antigen at four-week intervals. This delayed hypersensitivity test was assessed by the increase in skin thickness. The activity of the antigen was assessed in five animals which had a previous natural BHV-1 infection with clinical signs and seroconversion. Anti-BHV-1 antibodies were detected by seroneutralisation and an enzyme-linked immunoassay. Only one animal showed a significant but slight increase in skin thickness after the first test, but it was negative after a second test. The animals remained seronegative after the first test. Seroconversion was identified in 11 of the 43 animals (25 per cent) submitted to repeated delayed hypersensitivity tests. Five of 37 animals seroconverted after only two tests. The serological response was transient in seven of 11 seroconverted calves. Repeated hypersensitivity tests were therefore able to induce a serological response in seronegative calves but the response was weak and often transient. The test must therefore be applied cautiously to seronegative animals.

Animals↗

Location and characterization of the bovine herpesvirus type 4 thymidine kinase gene; comparison with thymidine kinase genes of other herpesviruses.

The location and nucleotide sequence of the bovine herpesvirus type 4 (BHV-4) thymidine kinase (TK) gene was determined. The coding region of the TK gene is 1335 nucleotides long and corresponds to a polypeptide of 445 amino acids. Comparison of TK amino acid sequences of BHV-4 and 16 herpesvirus TKs reveals a greater homology to those of the gammaherpesviruses EBV and specially HVS, than to those of alphaherpesviruses. The open reading frames detected in the vicinity of TK gene were homologous to the corresponding ones in other herpesviruses.

Amino Acid Sequence↗

Genetic relationships between bovine herpesvirus 4 and the gammaherpesviruses Epstein-Barr virus and herpesvirus saimiri.

The overall arrangement of genes in the unique central part of the bovine herpesvirus type 4 (BHV-4) genome has been deduced by analysis of short DNA sequences. Twenty-three genes conserved in at least one of the completely sequenced herpesviruses have been identified and localized. All of these genes encoded amino acid sequences with higher similarity to proteins of the gammaherpesviruses Epstein-Barr virus (EBV) and herpesvirus saimiri (HVS) than to the homologous products of the alphaherpesviruses varicella-zoster virus and herpes simplex virus type 1 or the betaherpesvirus human cytomegalovirus. The genome organization of BHV-4 had also an overall colinearity with that of the gammaherpesviruses EBV and HVS. Furthermore, the BHV-4 genes content and arrangement were more similar to those of HVS than to those of EBV, suggesting that BHV-4 and HVS are evolutionarily more closely related to each other than either are to EBV. BHV-4 DNA sequences were generally deficient in CpG dinucleotide. This CpG deficiency is characteristic of gammaherpesvirus genomes and suggests that the BHV-4 latent genome is extensively methylated. Despite several biological features similar to those of betaherpesviruses, BHV-4 displays the molecular characteristics of the representative members of the gammaherpesvirinae subfamily.

Amino Acid Sequence↗

Epidemiological evaluation of a monoclonal ELISA detecting bovine viral diarrhoea pestivirus antigens in field blood samples of persistently infected cattle.

An enzyme-linked immunosorbent assay (ELISA), using monoclonal antibodies for capture and detection, was developed for detecting bovine viral diarrhoea virus (BVDV) antigens in blood samples. The test was evaluated using 761 field samples of known status (viraemic or not). When an appropriate cut-off value was chosen, the sensitivity, specificity, and predictive values of the assay were 100%, higher than the values obtained by classical virus isolation. Correlation with the latter technique exceeded 90%. The ELISA is a good candidate for replacing virus isolation as a reference method for BVDV antigen detection in persistently infected carriers. A method based on the mean of the standard deviation ratio can be used to choose the cut-off value in order to optimise reproducibility.

Animals↗

Molecular biology of bovine herpesvirus type 4.

Bovine herpesvirus type 4 (BHV-4) is a ubiquitous virus of cattle. Its genome is a 144 +/- 6 kb double-stranded DNA consisting of a unique central part (L-DNA) flanked at both ends by tandem repeats called polyrepetitive DNA (prDNA or H-DNA). The overall arrangement of genes has been obtained by the analysis of homologies between short BHV-4 DNA sequences and corresponding genes of Epstein-Barr virus (EBV) and herpesvirus saimiri (HVS). The gene expression is temporally regulated. Glycoprotein precursor p (gp10/gp17) is expressed as gamma 1 polypeptide. Glycoproteins gp1, gp8, gp11 and their precursors are gamma 2 proteins. The analysis of strain variations allows the definition of two types of strains, based on the DNA patterns: the Movar 33/63-like and the DN 599-like strains. Only the M40 strain, isolated in India, fails to fit this classification. The genomic variations have been compiled to build a dendrogram showing three levels of divergence between BHV-4 strains or isolates. The available molecular data indicate that the BHV-4 genome shares much similarity with the DNA of EBV and HVS, two representative members of the gammaherpesvirinae. BHV-4 may therefore be classified in the subfamily gammaherpesvirinae.

Animals↗

Proteins of bovine herpesvirus type 4 released into the culture medium of productively infected cells: identification of a 135K glycoprotein involved in viral attachment.

Three bovine herpesvirus type 4 (BHV-4) proteins released into the culture medium of infected cells were identified, with Mr values of 135K, 16K and 14.5K. Among these three proteins, two were precipitated by the monoclonal antibodies characterized in this work. One is a glycoprotein of 135K (gp8) which does not seem to be involved in BHV-4 neutralization. Moreover, this 135K glycoprotein adsorbed onto uninfected susceptible cells. The attachment of gp8 to cells was totally inhibited by the prior adsorption of unlabelled viral proteins. Moreover, anti-gp8 monoclonal antibodies were effective in inhibiting the adsorption of gp8. These results indicate that gp8 could be involved in BHV-4 attachment.

Animals↗

Identification and characterization of glycoprotein gp1 of bovine herpesvirus type 4.

Three major bovine herpesvirus type 4 (BHV-4) glycoproteins have been described previously. By using monoclonal antibodies produced against BHV-4 envelope proteins from which the three major antigens had been removed by immunoaffinity, a fourth glycoprotein was identified. This protein (gp1) has a high Mr (greater than 300K), is detected about 8 h post-inoculation of infected cells and is strictly expressed as a gamma protein. Moreover, gp1 was identified by a polyclonal antiserum from an infected animal, indicating that this glycoprotein is an antigen recognized by the immune system of infected animals.

Animals↗

Genomic diversity among bovine herpesvirus 4 field isolates.

Twenty-eight Belgian field isolates of bovine herpesvirus 4 (BHV-4) coming from a variety of clinical diseases have been studied by restriction analysis and Southern blot hybridization. The unique central part of the genome was very well conserved among strains; only one variation in a restriction site was detected in 3 isolates which contain an additional EcoRI site also present in the LVR 140 strain; three regions in the unique part of the genome varied in size, one of these was highly variable. The polyrepetitive fragments (prDNAs) situated in tandem at both genomic ends were also variable in size; most of the isolates exhibited prDNA units of one size (major prDNA) and some of them also contained prDNA units having a different size and present in a lower amount (minor prDNA) than the major prDNA. Other isolates possessed two major prDNAs of different sizes which were both present in the same genome. The left junction fragment between the unique and the repeated sequences was also highly variable. No relationship could be established between the restriction pattern and the origin of the isolates; patterns of isolates coming from the same herd were similar except in one case. This study provides a view of the genome variability existing between BHV-4 field isolates.

Animals↗

Bovine herpesvirus 4 isolates: a comparison of three major glycoproteins.

Twenty-four Belgian field isolates of bovine herpesvirus 4 (BHV-4), together with four reference strains were compared by radio-immunoprecipitation and western blotting using a polyvalent antiserum and monoclonal antibodies raised against major glycoproteins. Most of these strains showed the same protein profile as the European reference strain Movar 33/63. For two strains the molecular weight of gp 6, p (gp 10/gp 17) and gp 10 were the same as those of the American reference strain DN 599. No relationship could be established between the protein profiles and origin of the isolates or with the restriction patterns. This study provides a view of the molecular weight variations of the major BHV-4 glycoproteins among field isolates.

Animals↗

A mathematical model of rinderpest infection in cattle populations.

A mathematical model for the epidemiology of rinderpest was developed, starting from a simplified descriptive analysis of the disease. A formula for the calculation of the probability of infection of a susceptible animal was first established. A deterministic failure threshold of the infection was then deduced. Deterministic and stochastic approaches were adopted using iterative methods on a computer. These allowed a description of the spread and the variability of an infection process in a population to be made. An illustration of the use of this model showed that, in some cases, variability effects due to stochastic factors were very important. In these particular conditions, the use of the deterministic model alone was not adequate for a good description of the infection. Consequently, improvements of the model were proposed in order to make it more realistic and to allow its use for the evaluation of the efficiency of field operations.

Animals↗

Antigenic and genomic identity between simian herpesvirus aotus type 2 and bovine herpesvirus type 4.

Herpesvirus aotus type 2 (HVA-2) was isolated from a culture of kidney cells from a healthy owl monkey (Aotus trivirgatus). Bovine herpesvirus type 4 (BHV-4) is frequently isolated from diseased and even healthy cattle and occasionally from sheep, wild ruminants and cats. The two viruses are related antigenically, as was revealed by an indirect fluorescent antibody test using polyclonal antisera from experimentally infected rabbits or monoclonal antibodies raised against six BHV-4 proteins, three of which were glycosylated. The genome structures of the two viruses consist of a unique central sequence flanked at both ends by G + C-rich tandem repeats. Restriction maps (produced using EcoRI, BamHI and HindIII) of these two viruses were nearly identical but the unique sequence of the HVA-2 genome possessed two additional BamHI sites. Four genomic regions of variable size were detected, two located in the unique part, one in the repetitive part and one in the left junction between the unique and the repeated part of the genome; these slight variations were similar to those observed between various BHV-4 isolates. These results suggest that HVA-2 and BHV-4 belong to the same virus species; HVA-2 could be either a BHV-4 contaminant of owl monkey kidney cell cultures or an isolate from an owl monkey accidentally infected with BHV-4.

Animals↗