Rapid detection of bovine leukaemia virus infection in a large cattle population with an ELISA performed on pooled sera grouped by herd.
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Biomedical subjects
Publications and source records attributed to M Mammerickx.
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The bovine leukemia virus is the etiological agent of a chronic lymphatic leukemia in cows, sheep, and goats. The same virus seems to induce a kind of wasting disease in experimentally infected rabbits. Antibodies to highly purified bovine leukemia viral Mr 51,000 glycoprotein and Mr 24,000 protein cross-react with human T-lymphotropic virus III/lymphadenopathy-associated virus antigens present in cultured lymphocytes of African patients suffering from acquired immune deficiency syndrome. Bovine leukemia virus has many structural and functional characteristics in common with the human T-lymphotropic viruses. The most striking feature of these retroviruses is the existence of a long open reading frame located at the 3' side of the provirus between the right end of the 3' side of env gene and the left end of the long terminal repeat. It is believed that the long open reading frame protein product acts in trans upon a number of genes to account for the biological effects of the virus.
Infection of bovines with bovine leukaemia virus (BLV) manifests itself in either of two ways: 30-70% of carriers develop persistent lymphocytosis (PL), with the viral genome integrated at a large number of different sites in the DNA of the affected B-lymphocytes, without causing any chromosomal abnormalities. Only 0,1-10% of carriers develop lymphoid tumours, which also consist of B-lymphocytes. In contrast to PL, however, they are of mono- or oligoclonal origin in terms of the integration site, which is characteristic for each tumour. All cells contain one or more copies of the viral genome, chromosomal aberrations are common and if deletions are present they are invariably found in the 5'-half of the virus DNA sequence. In both types of affected cells transcription is repressed in vivo, but transient virus production can be induced in vitro and detected by means of syncytia induction or haemagglutination. In vivo production of virus in some unknown cell is suggested by the presence of high antibody titres in infected animals, especially against the envelope glycoprotein gp51. This can be detected by various techniques such as immunodiffusion, radioimmune assay or ELISA. Monoclonal antibodies against gp51 have revealed 8 epitopes, 3 of which are recognized by neutralizing antibodies and one by a cytolytic antibody. The BLV genome, about 9 kb in size, have been cloned, and some of the information obtained on its molecular structure and function is discussed. It codes for at least 4 non-glycosylated and 2 glycoproteins. Of special interest is the recently discovered serological relationship between some of the non-glycosylated proteins and those of the human T-cell leukaemia virus. The functional role of BLV in leukaemogenesis is largely unknown. The presence of the viral genome seems to be necessary for the maintenance of the transformed state, but not its continuous expression nor an LTR-mediated promotion of transcription of cellular genes. No oncogene is carried by the virus. Although bovine leukosis is not of major economic importance, its eradication is desirable and feasible in countries with a relatively low incidence, by means of testing and elimination. For endemic situations vaccination would be preferable, and distinct possibilities exist for the development of gp51 based vaccines.
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A panel of monoclonal antibodies to the bovine leukemia virus envelope glycoprotein (BLV gp51) has previously demonstrated the association of the biological activities of the virus (infectivity, syncytia induction) with three out of eight epitopes of gp51. In BLV-infected cells, the unglycosylated homolog of the precursor to the BLV envelope glycoproteins (gPr72env) is a 47,000-MW polypeptide. Immunoprecipitation studies with monoclonal antibodies show that the neutralizing antibody-inducing sites, although present in gPr72env, are not conserved in the 47,000-MW unglycosylated homolog. Finally, it is demonstrated that the neutralizing antibody-inducing sites of gp51 are subject to antigenic variation among BLV isolates of the same or different geographical origins.
Bovine leukemia virus (BLV) proviral integration was studied in the DNA from circulating leucocytes or tumor cells of sheep and goats experimentally infected with BLV. Southern blot analysis of infected cell DNA for BLV proviral sequences indicate that: (1) the provirus may be found as unintegrated molecules in the circulating leucocytes of infected sheep; (2) the provirus is integrated at many sites in the genome of the leucocytes of infected goats and occasionally in infected sheep; (3) the provirus is present at only a few sites in the DNA of sheep or goat tumor cell clones. A second case of goat lymphosarcoma is also reported.
Sera of BLV-infected cattle and sheep are tested for their reactivity with different gp51 subregions by competition with radiolabeled monoclonal antibodies directed against 8 different gp51 epitopes. Sheep antisera are found to be very polyvalent, since they are able to displace the fixation of any of the mouse monoclonal antibodies to gp51. Bovine antisera do not display significant competition with monoclonal antibodies direct against 5 out of 8 gp51 epitopes. The bovine antibody response to gp51 is focused to a limited subregion of this molecule, bearing 3 epitopes (F, G and H) recognized by antibodies with virus-neutralizing activity. The differential reactivity of cattle and sheep antisera to BLV gp51 is discussed in relation to the pathology of BLV infection in these two species.
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Bovine leukemia virus (BLV) has many structural and functional characteristics in common with the human T-lymphotropic viruses (HTLVs). The most striking feature of these retroviruses is the existence of a long open reading frame (LOR) located at the 3' side of the provirus between the right end of the 3' side of env gene and the left end of the long terminal repeat (LTR). It is believed that the LOR protein product is of critical importance in the induction process of the tumor phase of bovine leukemia. Prevention of BLV infection will be attempted by vaccination. To that aim, careful study of BLV envelope glycoprotein epitopes has shown that epitopes F, G, and H play a major role in biological activities of the virus. Their native structure depends upon glycosylation of the peptide backbone.
A variant of the ELISA technique, involving a monoclonal anti-gp51 antibody yields a highly sensitive method for the detection of bovine leukaemia virus (BLV) antibodies. The gp51 antigen-coated microtitre plates are obtained by incubation of plastic-adsorbed monoclonal antibodies with a non-purified mixture of BLV antigens. Sera to be tested are incubated in the wells of the gp51-coated plates and bound antibodies are revealed by an enzyme-linked antibovine immunoglobulin reagent. This test is as sensitive as liquid phase radioimmunoassay using the same gp51 antigen and thus appears as a highly sensitive, practical, rapid and cheap method for the detection of BLV antibodies.
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The anticoccidial effect of narasin against Eimeria flavescens, E. intestinalis, E. magna, E. perforans and E. stiedai was tested in experimentally infected rabbits. The drug was highly effective in reducing oocyst output and in preventing clinical signs. Maximum activity as measured by oocyst discharge was achieved with feed levels from 12 to 24 p.p.m.; there was no influence on oocyst sporulation. Optimal weight gain and food consumption were obtained with levels between 8 and 12 p.p.m.; at higher levels there was an adverse effect on growth, food intake, relative liver and heart weights and on serum potassium and glucose.
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