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1.
Prev Vet Med ; 133: 1-9, 2016 Oct 01.
Article in English | MEDLINE | ID: mdl-27720022

ABSTRACT

Enzootic bovine leukosis (EBL) is an economically important disease of dairy cattle caused by bovine leukemia virus (BLV). The economic impacts of the infection have been debated in the literature. The present study was conducted to determine the lifetime effects of BLV infection on longevity and milk production of dairy cows in Canada. The data were aggregated from a combination of two data sets: 1) BLV serum-ELISA test results from Canada-wide surveys of production limiting diseases, which took place between 1998 and 2003 in 8 provinces, and 2) longitudinal production data for all cows in the former study, extracted from the Canadian dairy herd improvement database. All participant cows had been culled or died by the onset of this study. A historical cohort study was designed, including cows which tested positive to BLV-antibodies in their first lactation (positive cohort, n=1858) and cows which tested negative in their second or later lactations (negative cohort, n=2194). To assess the impacts of infection with BLV on longevity (the number of lifetime lactations), a discrete-time survival analysis was carried out. The effect of BLV on the lifetime milk production (the sum of all life 305-day milk production) was evaluated using a multilevel linear regression model. Overall, 4052 cows from 348 herds met the eligibility criteria and were enrolled in the study. In the longevity model, the interaction term between time (lactation number) and BLV-status was highly significant. Cows which were positive to BLV had consistently greater probabilities of being culled (or dying) than the test-negative cows. In the milk production model, the interaction term between BLV-status and longevity of the cows was highly significant; indicating that lifetime BLV effects on the total milk production was dependent on the lactation in which the study cows were culled/died. Infected cows with 2 and 3 lactations showed significantly lower life milk productions [-2554kg (-3609 to -1500) and -1171kg (-2051 to -292), respectively] compared with their negative counterparts with 2 and 3 lactations. As the cows lived longer (>3 lactations), the differences in life milk production between the two cohorts were no longer significant. Overall, it was predicted that the test-positive cows produced substantially lower milk compared to the test-negative cows throughout their study lifespans. With the high prevalence of BLV in Canadian dairy cows and its detrimental economic impacts, pursuing broad-based control programs in Canada should be evaluated.


Subject(s)
Enzootic Bovine Leukosis/physiopathology , Lactation/physiology , Leukemia Virus, Bovine/physiology , Longevity , Animals , Canada , Cattle , Cohort Studies , Enzootic Bovine Leukosis/microbiology , Female , Milk/metabolism , Milk/microbiology , Retrospective Studies
2.
Proteins ; 81(8): 1377-85, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23504872

ABSTRACT

The matrix (MA) domain of retroviral Gag proteins plays a crucial role in virion assembly. In human immunodeficiency virus type 1 (HIV-1), a lentivirus, the presence of phosphatidylinositol-(4,5)-bisphosphate triggers a conformational change allowing the MA domain to bind the plasma membrane (PM). In this study, the MA protein from bovine leukemia virus (BLV) was used to investigate the mechanism of viral Gag binding to the membrane during replication of a deltaretrovirus. Fluorescence spectroscopy was used to measure the binding affinity of MA for two RNA constructs derived from the BLV genome as well as for single-stranded DNA (ssDNA). The importance of electrostatic interactions and the ability of inositol hexakisphosphate (IP6) to compete with nucleic acids for binding to MA were also investigated. Our data show that IP6 effectively competes with RNA and DNA for BLV MA binding, while [NaCl] of greater than 100 mM is required to produce any observable effect on DNA-MA binding. These results suggest that BLV assembly may be highly dependent on the specific interaction of the MA domain with components of the PM, as observed previously with HIV-1. The mode of MA binding to nucleic acids and the implications for BLV assembly are discussed.


Subject(s)
Gene Products, gag/metabolism , Inositol Phosphates/metabolism , Leukemia Virus, Bovine/physiology , Nucleic Acids/metabolism , Animals , Cattle , Enzootic Bovine Leukosis/microbiology , Spectrometry, Fluorescence , Virus Replication
3.
Rev. MVZ Córdoba ; 13(1): 1197-1204, ene.-abr. 2008. tab, graf
Article in Spanish | LILACS | ID: lil-498569

ABSTRACT

Objetivo. Determinar la seroprevalencia de Leucosis Viral Bovina (LVB) en animales contrastornos reproductivos. Materiales y métodos. Se recolectaron 137 muestras de sangrede hembras con antecedentes de infertilidad, pertenecientes a 28 fincas distribuidas en elmunicipio de Montería; adicionalmente, se obtuvieron muestras al azar de 26 torospertenecientes a las mismas fincas que fueron analizadas para anticuerpos contra LVB. Latécnica serológica empleada fue la prueba de ELISA. Se realizó un análisis descriptivo tabulandola información con datos de seropositividad y seronegatividad obtenidos de cada animal; losresultados se interpretaron de acuerdo a las variables: raza, edad, sexo, zona, tipo deexplotación y evento o problema reproductivo detectado. Para determinar la asociaciónentre seropositividad y cada una de las variables se utilizó la prueba de χ2. Resultados. Laspruebas arrojaron una seroprevalencia del 21% para LVB. No se encontraron diferenciassignificativas de prevalencia asociadas a las variables raza, edad o estado reproductivo delos animales (p≥0.05), pero si entre la presencia de anticuerpos contra LVB y las variableszona, tipo de explotación y sexo. Conclusiones. Se demuestra la circulación del virus de laLVB en Montería, (Colombia). Se confirma la importancia de implementar un programa decontrol y prevención de la diseminación de la infección, con el fin de evitar las pérdidaseconómicas asociadas, y dentro de lo posible, la eliminación de los especímenes seropositivospara lograr la erradicación de la infección en esta zona del país.


Subject(s)
Cattle , Enzootic Bovine Leukosis , Reproduction , Viruses , Cattle/genetics , Cattle/blood , Enzootic Bovine Leukosis/microbiology , Enzootic Bovine Leukosis/blood , Reproduction/genetics , Viruses/immunology , Viruses/pathogenicity
4.
J Dairy Sci ; 90(2): 659-69, 2007 Feb.
Article in English | MEDLINE | ID: mdl-17235141

ABSTRACT

The primary purpose of this research was to determine associations among seropositivity for bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV), Mycobacterium avium ssp. paratuberculosis (MAP), and Neospora caninum (NC) and each of 3 outcome variables (305-d milk, fat, and protein production) in Canadian dairy cattle. Serum samples from up to 30 randomly selected cows from 342 herds on monthly milk testing were tested for antibodies against BLV (IDEXX ELISA; IDEXX Corporation, Westbrook, ME), MAP (IDEXX or Biocor ELISA; Biocor Animal Health, Inc., Omaha, NE), and NC (IDEXX or Biovet ELISA; Biovet Inc., St. Hyacinthe, Quebec, Canada). Up to 5 unvaccinated cattle over 6 mo of age were tested for virus-neutralizing antibodies to the Singer strain of type 1 BVDV. Dairy Herd Improvement records were obtained electronically for all sampled cows. Linear mixed models with herd and cow as random variables were fit, with significant restricted maximum likelihood estimates of outcome effects being obtained, while controlling for potential confounding variables. Bovine leukemia virus seropositivity was not associated with 305-d milk, 305-d fat, or 305-d protein production. Cows in BVDV-seropositive herds (at least one unvaccinated animal with a titer > or =1:64) had reductions in 305-d milk, fat, and protein of 368, 10.2, and 9.5 kg, respectively, compared with cows in BVDV-seronegative herds. Mycobacterium avium ssp. paratuberculosis seropositivity was associated with lower 305-d milk of 212 kg in 4+-lactation cows compared with MAP-seronegative 4+-lactation cows. Neospora caninum seropositivity in primiparous cows was associated with lower 305-d milk, fat, and protein of 158, 5.5, and 3.3 kg, respectively, compared with NC-seronegative primiparous cows. There were no interactions among seropositivity for any of the pathogens and their effects on any of the outcomes examined, although the low MAP seroprevalence limited this analysis. Results from this research will contribute to understanding the economic impacts of these pathogens and justify their control.


Subject(s)
Bovine Virus Diarrhea-Mucosal Disease/physiopathology , Coccidiosis/veterinary , Enzootic Bovine Leukosis/physiopathology , Lactation/physiology , Paratuberculosis/physiopathology , Animals , Antibodies, Bacterial/blood , Antibodies, Protozoan/blood , Antibodies, Viral/blood , Bovine Virus Diarrhea-Mucosal Disease/microbiology , Canada , Cattle , Coccidiosis/parasitology , Coccidiosis/physiopathology , Diarrhea Viruses, Bovine Viral/immunology , Enzootic Bovine Leukosis/microbiology , Female , Leukemia Virus, Bovine/immunology , Milk/chemistry , Milk Proteins/analysis , Mycobacterium/immunology , Neospora/immunology , Paratuberculosis/microbiology , Pregnancy
5.
J Immunol Methods ; 199(2): 159-65, 1996 Dec 15.
Article in English | MEDLINE | ID: mdl-8982358

ABSTRACT

Evaluation of antigen specific lymphocyte proliferation is extremely difficult with cells displaying high spontaneous proliferation in vitro, such as bovine leukemia virus (BLV) and T cell leukemia virus type I (HTLV-I) infected cells. We introduced modifications to a standard proliferation protocol, and for the first time report antigen specific proliferation in BLV infected animals with persistent lymphocytosis (a benign proliferation of B lymphocytes). Serum (autologous, heterologous or fetal) but not medium was a major factor contributing to spontaneous proliferation. Spontaneous proliferation was strongly serum concentration dependent and reducing serum to 0.5-1% resulted in decreased background proliferation and development of a functional assay. Addition of indomethacin (3-6 micrograms/ml) further increased antigen specific responses not only from persistently lymphocytotic but also from lymphosarcoma animals. Thus, this modified proliferation protocol can be used in infections where background proliferation hinders evaluation of antigen-specific CD4+ T cells.


Subject(s)
CD4-Positive T-Lymphocytes/immunology , Enzootic Bovine Leukosis/microbiology , Lymphocyte Activation , Animals , Cattle , Cyclooxygenase Inhibitors/pharmacology , Indomethacin/pharmacology , Leukemia Virus, Bovine , Lymphocyte Depletion
6.
Cancer Lett ; 86(1): 33-40, 1994 Oct 28.
Article in English | MEDLINE | ID: mdl-7954352

ABSTRACT

Fetal lamb kidney cells (FLK) and bat lung (BAT2CL6) cells that continuously produce bovine leukemia virus (BLV) were found to cause malignant tumors in nude mice. Uninfected bat lung cells (Tb 1 Lu) produced a small benign neoplasm. Pulse chase studies showed that the p53 gene product in BAT2CL6 cells was more stable compared with p53 in Tb 1 Lu cells. Mono-clonal antibody studies suggested that a mutant form of the p53 protein was produced in BLV-infected cells. Heteroduplex mapping studies of the p53 gene from BLV-infected cells also suggested that a mutation in p53 had occurred. Stabilization of the p53 gene product in BLV-infected cells may contribute to the progression of tumor virulence.


Subject(s)
Leukemia Virus, Bovine , Neoplasms, Experimental/virology , Tumor Suppressor Protein p53/metabolism , Animals , Cattle , Cells, Cultured , Chiroptera , Chromosome Mapping , Drug Stability , Enzootic Bovine Leukosis/microbiology , Genes, p53 , Kidney/cytology , Kidney/microbiology , Lung/cytology , Lung/microbiology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Nude , Neoplasms, Experimental/pathology , Nucleic Acid Heteroduplexes , Radioimmunoprecipitation Assay , Sheep , Tumor Suppressor Protein p53/genetics
7.
Lab Invest ; 71(3): 359-65, 1994 Sep.
Article in English | MEDLINE | ID: mdl-7933986

ABSTRACT

BACKGROUND: Bovine leukemia virus (BLV), a retrovirus, usually causes a subclinical infection of dairy and beef cattle, but in < 1% of infected cattle a B cell lymphoma may develop after several years of infection. BLV is transmitted horizontally among cattle, and infected animals have anti-BLV titers. Expression of BLV antigen, however, is silent in peripheral blood lymphocytes in vivo. The tropism of BLV has been assumed to be limited to B lymphocytes, because no other cell type has been found to harbor the virus in vivo. Since retrovirus-like particles had been identified in milk, and infection can be transmitted by milk, we decided to investigate whether BLV was in mammary epithelial cells. EXPERIMENTAL DESIGN: Pure cultures of mammary epithelial cells were established from cells shed into the milk of 28 cows. BLV was searched in these cultures by immunocytochemistry and the polymerase chain reaction, the specificity of the latter confirmed by Southern blot hybridization and DNA sequencing. BLV was searched immunocytochemically in mammary tissue sections from 12 cows. RESULTS: Antigenic and/or molecular evidence of BLV was found in the cultured cells of 20 cows. Antigenic evidence of BLV was found in tissue sections from 10 cows, indicating virus expression in vivo. Simultaneous detection of BLV p24 and cytokeratins, localized antigen expression to the mammary epithelial cells. CONCLUSIONS: These indications of BLV in mammary epithelial cells in vivo suggest that BLV is capable of infecting and expressing antigen in glandular epithelium in vivo and has a broader tissue tropism than was previously thought. They raise the question of how persistent mammary infection by BLV may influence the course of bovine lymphoma, and what effect the virus may have on the mammary gland.


Subject(s)
Enzootic Bovine Leukosis/microbiology , Leukemia Virus, Bovine/isolation & purification , Mammary Glands, Animal/microbiology , Animals , Antibodies, Viral/analysis , Antigen-Presenting Cells/pathology , Antigens, Viral/analysis , Antigens, Viral/genetics , Blotting, Southern , Cattle , Cells, Cultured , DNA, Viral/analysis , Enzootic Bovine Leukosis/pathology , Epithelium/immunology , Epithelium/microbiology , Epithelium/pathology , Female , Immunohistochemistry , Leukemia Virus, Bovine/immunology , Lymphocytes/immunology , Mammary Glands, Animal/immunology , Mammary Glands, Animal/pathology , Milk/cytology , Milk/microbiology , Polymerase Chain Reaction
8.
J Virol ; 68(7): 4589-96, 1994 Jul.
Article in English | MEDLINE | ID: mdl-8207833

ABSTRACT

Bovine leukemia virus (BLV), an oncovirus related to human T-cell leukemia virus type I, causes a B-cell lymphoproliferative syndrome in cattle, leading to an inversion of the T-cell/B-cell ratio and, more rarely, to a B-cell lymphosarcoma. Sheep are highly sensitive to BLV experimental infection and develop B-cell pathologies similar to those in cattle in 90% of the cases. BLV tropism for B cells has been well documented, but the infection of other cell populations may also be involved in the BLV-induced lymphoproliferative syndrome. We thus looked for BLV provirus in other leukocyte populations in sheep and cattle by using PCR. We found that while B cells harbor the highest proviral load, CD8+ T cells, monocytes, and granulocytes, but not CD4+ T cells, also bear BLV provirus. As previously described, we found that persistent lymphocytosis in cows is characterized by an expansion of the CD5+ B-cell subpopulation but we did not confirm this observation in sheep in which the expanded B-cell population expressed the CD11b marker. Nevertheless, BLV could be detected both in bovine CD5+ and CD5- B cells and in sheep CD11b+ and CD11b- B cells, indicating that the restricted BLV tropism for a specific B-cell subpopulation cannot explain its expansion encountered in BLV infection. Altogether, this work shows that BLV tropism in leukocytes is wider than previously thought. These results lead the way to further studies of cellular interactions among B cells and other leukocytes that may intervene in the development of the lymphoproliferative syndrome induced by BLV infection.


Subject(s)
Leukemia Virus, Bovine/isolation & purification , Leukocytes/microbiology , Animals , B-Lymphocytes/metabolism , Base Sequence , Cattle , Cell Line , DNA Primers , Enzootic Bovine Leukosis/microbiology , Lymphocyte Subsets/microbiology , Lymphocytosis/pathology , Molecular Sequence Data , Proviruses/isolation & purification , Sheep
9.
Leukemia ; 8(6): 1057-61, 1994 Jun.
Article in English | MEDLINE | ID: mdl-7516028

ABSTRACT

Peripheral blood lymphocytes from bovine leukemia virus (BLV)-negative and BLV-infected, aleukemic cows with persistent lymphocytosis were evaluated for expression of B and T lymphocyte subset-specific molecules and co-expression of the interleukin-2 receptor alpha (IL-2R alpha) molecule. Results demonstrate enhanced mitogen-induced expression of the IL-2R alpha molecule on B lymphocytes from BLV-infected, lymphocytotic cows. Lymphocyte subset analyses further demonstrate that BLV-infected, lymphocytotic cows are not only characterized by sustained elevations in CD5+ B lymphocytes, but also show significantly elevated numbers of CD3+, CD4+, and CD8+ T lymphocytes. These results provide evidence suggesting that B lymphocytes from BLV-infected, lymphocytotic cows are more sensitive to activation signals and up-regulation of the IL-2 signaling pathway than lymphocytes from clinically normal BLV-free cows, and that T lymphocytes may be involved in the aberrant regulatory pathways underlying BLV-induced persistent B lymphocytosis.


Subject(s)
B-Lymphocytes/physiology , Enzootic Bovine Leukosis/blood , Leukemia Virus, Bovine , Lymphocytosis/blood , Receptors, Interleukin-2/physiology , T-Lymphocytes/physiology , Animals , Antigens, CD/analysis , B-Lymphocytes/immunology , B-Lymphocytes/ultrastructure , CD4-CD8 Ratio , CD4-Positive T-Lymphocytes/physiology , CD5 Antigens , Cattle , Enzootic Bovine Leukosis/microbiology , Female , Leukocyte Count , Lymphocyte Subsets/immunology , Lymphocytosis/immunology , Phenotype , T-Lymphocytes/cytology , T-Lymphocytes/immunology
10.
Vet Microbiol ; 39(3-4): 313-21, 1994 Apr.
Article in English | MEDLINE | ID: mdl-8042277

ABSTRACT

A polymerase chain reaction (PCR) procedure that detects proviral bovine leukaemia virus (BLV) in peripheral blood mononuclear cell DNA was evaluated. Blood samples from all animals (164) in a commercial dairy herd with a 30% prevalence of BLV infection, and from 194 animals from BLV free herds were tested. The absence of any positive PCR results in animals from BLV free herds confirmed the specificity of the assay. Initial testing of the infected herd using a single amplification PCR (SA-PCR), detected BLV infection in 62 of 72 adult animals that were seropositive by the agar gel immunodiffusion (AGID) test and in one persistently seronegative cow. Infection in this cow was confirmed by sheep bioassay. Subsequent testing of the SA-PCR negative, seropositive animals using a double amplification PCR (DA-PCR) detected proviral BLV in eight of nine animals that were available for retesting. The PCR assay was also able to distinguish BLV infected calves from uninfected calves that were serologically positive because of the presence of colostral antibody. Lymphocytes from all seropositive animals were cultured for determination of BLV antigen expression. Cultures from 37 of 62 SA-PCR positive animals produced detectable quantities of viral antigens. However, antigen expression was not detected in cultures from seropositive animals that were negative in the SA-PCR. In addition, in experimental transmission tests, inoculation of more than 10(6) lymphocytes from these cows was required for sheep to become seropositive to BLV.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
DNA, Viral/blood , Enzootic Bovine Leukosis/microbiology , Leukemia Virus, Bovine/genetics , Polymerase Chain Reaction , Proviruses/genetics , Animals , Base Sequence , Cattle , DNA Primers/chemistry , DNA, Viral/chemistry , Enzootic Bovine Leukosis/transmission , Evaluation Studies as Topic , Immunodiffusion , Leukemia Virus, Bovine/isolation & purification , Leukocytes/microbiology , Molecular Sequence Data , Proviruses/isolation & purification , Sheep
11.
Leuk Res ; 18(2): 91-9, 1994 Feb.
Article in English | MEDLINE | ID: mdl-8107493

ABSTRACT

Despite detailed knowledge of the genetic map of the bovine leukemia virus (BLV), the mechanism whereby BLV infection results in transformation and B-lineage restriction of tumors is poorly understood. The aim of this study was to gain new insight into pathogenetic mechanisms of BLV-induced tumorigenesis by determining the karyotypes of BLV-associated lymphomas in cattle. Metaphases in cells from lymphoid tumors from 20 mature dairy cows were banded and analyzed after short-term, unstimulated culture. Nineteen out of twenty cases exhibited clonal abnormalities, 17 cases were hyperdiploid, and 16 cases had extremely complex chromosomal changes. Recurrent chromosomal anomalies were identified and there was clear evidence for the evolution of increasing chromosomal instability in 12 cases. The most common abnormalities were the acquisition of additional small chromosomes (23-29); trisomy of chromosomes 5 and 7, and Robertsonian translocations and isochromosome rearrangements involving chromosomes 10, 12, 23, and 26. Monosomy X, trisomy X, and translocations involving the X chromosome were also detected. Chromosomes 2, 3, 4, 6, 8, 9, 11, 13, 14, 19, and 21 were infrequently involved in either structural or numerical changes. Structural rearrangements of chromosomes 10, 12, 23, and 26 may reflect primary abnormalities occurring relatively early in transformation, whereas trisomy 5 may be an extremely common secondary abnormality. While comparison of these findings with the current bovine gene map raises intriguing possibilities for pathogenetic mechanisms, further studies are needed before hypothetical mechanisms linking chromosomal abnormalities with BLV-induced transformation can be made.


Subject(s)
Chromosome Aberrations , Enzootic Bovine Leukosis/genetics , Animals , B-Lymphocytes/microbiology , B-Lymphocytes/pathology , Cattle/genetics , Cell Transformation, Neoplastic/genetics , Cell Transformation, Viral/genetics , Enzootic Bovine Leukosis/microbiology , Female , Karyotyping/veterinary , Translocation, Genetic , Trisomy
12.
Am J Vet Res ; 55(1): 72-80, 1994 Jan.
Article in English | MEDLINE | ID: mdl-8141500

ABSTRACT

We used monoclonal antibodies and immunohistologic examination of lymph nodes, to elucidate the pathogenesis of lymphosarcoma induced by infection with bovine leukemia virus (BLV). The superficial cervical lymph nodes from 3 BLV-infected but apparently healthy sheep and 5 sheep with full-blown lymphosarcoma were examined. We also investigated the integration of bovine leukemia provirus by use of Southern blotting. In lymph nodes from sheep lacking clinical signs of infection, in which the provirus had been integrated at multiple sites in the genome, many large hypertrophic follicles were observed in the cortex. These follicles had germinal centers consisting of CD4+T cells and B cells that expressed surface IgM (sIgM) and major histocompatibility complex (MHC) class-II antigens, but not B cell-specific B2 molecule. The percentage of CD4+T cells in the cortex was significantly (P < 0.05) higher than that of the controls and sheep with lymphosarcoma. In all sheep with lymphosarcoma, the lymph nodes were completely destroyed by proliferating neoplastic cells, and in addition, small atrophic follicles, which consisted of normal B-cell marker-positive cells, were seen near the trabecula and the subcapsule. In these instances, neoplastic cells appeared to be a monoclonal population derived from a single CD5- B-cell lineage and to be classified as 2 types, CD5-CD4-CD8-B2+MHC class-II+sIgM+ and CD5-CD4-CD8-B2+MHC class-II+sIgM-. Moreover, CD8+T cells infiltrated diffusely throughout the tumorous lymph nodes apart from the atrophic follicles, and CD4+ cells were observed around atrophic follicles. Both types of T cells were small-size, normal lymphocytes with round and noncleaved nuclei, and were apparently non-neoplastic cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Leukemia Virus, Bovine/pathogenicity , Lymphocytes/immunology , Lymphoma, Non-Hodgkin/microbiology , Animals , Antibodies, Monoclonal , Antibodies, Viral/blood , B-Lymphocytes/immunology , Cattle , Cell Transformation, Neoplastic , DNA, Viral/isolation & purification , Enzootic Bovine Leukosis/microbiology , Female , Immunoglobulin M/biosynthesis , Leukemia Virus, Bovine/isolation & purification , Lymph Nodes/immunology , Lymph Nodes/microbiology , Lymphocytes/microbiology , Lymphoma, Non-Hodgkin/blood , Lymphoma, Non-Hodgkin/immunology , Male , Sheep , T-Lymphocyte Subsets/immunology , T-Lymphocytes/immunology
13.
J Virol ; 67(9): 5321-7, 1993 Sep.
Article in English | MEDLINE | ID: mdl-7688821

ABSTRACT

A battery of 19 synthetic peptides was used to characterize efficient neutralizing and helper T-cell epitopes on the bovine leukemia virus (BLV) external envelope glycoprotein gp51. Four of the antipeptide antisera raised in rabbits inhibited the formation of BLV-induced syncytia; these antisera are directed against peptides 64-73, 98-117, and 177-192. Only antisera directed against the 177-192 region also neutralized vesicular stomatitis virus-BLV pseudotypes. This study clearly demonstrates that neutralizing properties can be observed with antibodies raised to regions undescribed so far and included in both the amino-terminal and central parts of the antigen. In addition, some helper T-cell determinants were defined from gp51-immunized mice and from BLV-infected cattle. Although none of the peptides tested behaved as a universal helper T-cell epitope, peptide 98-117 stimulated T-cell proliferation from BALB/c mice and from three infected cows, while peptide 169-188 strongly stimulated T-cell proliferation from one infected cow. Further experiments performed with three peptides overlapping the 169-188 region (177-192, 179-192, 181-192) demonstrated the particular relevance of residue(s) P-177 and/or D-178 in the helper T-cell epitope. These data should assist in the design of an efficient subunit vaccine against BLV infection that contains peptides possessing both B-neutralizing and helper T-cell determinants.


Subject(s)
B-Lymphocytes/immunology , Epitopes/immunology , Leukemia Virus, Bovine/immunology , T-Lymphocytes, Helper-Inducer/immunology , T-Lymphocytes/immunology , Viral Envelope Proteins/immunology , Amino Acid Sequence , Animals , Cattle , Enzootic Bovine Leukosis/microbiology , Epitopes/analysis , Female , Giant Cells/physiology , Immune Sera , In Vitro Techniques , Leukemia Virus, Bovine/isolation & purification , Lymphocyte Activation , Mice , Mice, Inbred BALB C , Mice, Inbred C3H , Mice, Inbred C57BL , Molecular Sequence Data , Neutralization Tests , Peptides/chemical synthesis , Peptides/immunology
15.
Am J Vet Res ; 54(2): 205-9, 1993 Feb.
Article in English | MEDLINE | ID: mdl-8381625

ABSTRACT

A study was performed to determine whether experimentally induced bovine leukemia virus (BLV) infection in cattle could be detected earlier by use of polymerase chain reaction (PCR) amplification of genomic DNA extracted from leukocytes than by use of conventional agar-gel immunodiffusion (AGID). The PCR primers were designed to amplify a 375-base-pair region of the proviral gag gene. Five cows were identified that were BLV-negative on the basis of AGID and PCR results. At day 0, these cows were inoculated IM with blood pooled from 3 naturally infected cows. Blood samples were taken on days 0, 1, and 7, and every 2 weeks thereafter until 3 months after inoculation. Three of the cows were BLV-positive by AGID test results 3 weeks after inoculation, and the remaining 2 seroconverted at 5 weeks. In contrast, all 5 cows were BLV-positive by PCR results 7 days after inoculation and remained positive for the duration of the study. Five cows that were BLV-positive by AGID test and PCR results on day 0 and from which samples were obtained at the same times as those from the other 5 cows, remained BLV-positive by results of both tests during the course of the study. Results indicate that under experimental conditions, BLV infection in cattle can be detected as much as 2 to 4 weeks earlier by use of PCR than by use of the AGID test.


Subject(s)
DNA, Viral/genetics , Enzootic Bovine Leukosis/microbiology , Leukemia Virus, Bovine/isolation & purification , Polymerase Chain Reaction/veterinary , Animals , Base Sequence , Cattle , Enzootic Bovine Leukosis/diagnosis , Immunodiffusion/veterinary , Molecular Sequence Data
16.
Comp Immunol Microbiol Infect Dis ; 16(1): 63-71, 1993 Jan.
Article in English | MEDLINE | ID: mdl-8382592

ABSTRACT

Calves of two breeds--Brown Latvian and Black and White, were immunized against bovine leukemia virus with an immunogen produced from short-term cultured blood lymphocytes of BLV-positive Brown Latvian cows. The immunogen contained the viral proteins gp51 and p24 as well as some cellular proteins. As determined by bioassay in sheep, it did not contain infective bovine leukemia virus. The calves were challenged by BLV-producing lymphocytes 2 weeks after the immunization procedure. Effective protection against BLV infection was induced by the tested immunogen in all Brown Latvian calves for 6 months and in 70% of the Black and White calves.


Subject(s)
Enzootic Bovine Leukosis/prevention & control , Leukemia Virus, Bovine , Animals , Antibodies, Viral/blood , Antigens, Viral/administration & dosage , Antigens, Viral/isolation & purification , Cattle , Enzootic Bovine Leukosis/immunology , Enzootic Bovine Leukosis/microbiology , Evaluation Studies as Topic , Immunization , Leukemia Virus, Bovine/immunology , Leukemia Virus, Bovine/isolation & purification , Lymphocyte Activation , Lymphocytes/immunology , Lymphocytes/microbiology , Species Specificity , Viral Proteins/administration & dosage , Viral Proteins/immunology , Viral Proteins/isolation & purification
17.
Cancer Res ; 53(2): 429-37, 1993 Jan 15.
Article in English | MEDLINE | ID: mdl-8380256

ABSTRACT

The phenotype and ontogeny of cells carrying the tumor-associated antigen (TAA), identified in tumors of cattle with enzootic bovine leukosis (EBL) by use of the monoclonal antibody (MAb) c143, were analyzed by flow cytometry and immunohistochemistry. The TAA recognized by the c143 MAb (c143 TAA) was mainly expressed on B-cells, macrophages, reticular cells, and a minor population of BoCD4-positive T-cells in bovine leukemia virus (BLV)-free normal cattle. When the peripheral blood mononuclear cells from normal cattle were activated in vitro, some populations of BoCD8-positive T- and non-T/non-B-cells also showed expression. Moreover, B-cells expressed the c143 TAA until the antigen was lost from cells at the final stage of B-cell differentiation, namely, the plasma cell stage. The c143 MAb-positive cells in blood of BLV-free normal cattle form heterologous subpopulations, and these cells coexpress other surface markers such as BoCD2, BoCD5, BoCD6, and B-cell-specific molecules B1 low, B1 bright, and B2. In BLV-infected cattle, the proportion of peripheral blood mononuclear cells that express the c143 TAA increased with the progression of EBL, and, in addition, BLV-infected cattle that had no lymphosarcomas showed increased proportions of c143 MAb-positive cells that coexpressed surface IgM (sIgM), BoCD5, B1 low, and B2 but not BoCD2, BoCD4, or BoCD6. Furthermore, most of the c143 MAb-positive tumors from all cattle with EBL appeared to be a monoclonal population derived from a single B-cell and to be divided into two types, c143 TAA+BoCD5+B1 low+ B2+ sIgM+ or sIgM-. Collectively, these results show that the c143 TAA is not only a useful surface marker for identification of EBL but also a marker of differentiation of lymphoid cells.


Subject(s)
Antigens, Tumor-Associated, Carbohydrate/immunology , B-Lymphocytes/immunology , Enzootic Bovine Leukosis/immunology , Leukemia Virus, Bovine/pathogenicity , Animals , Antibodies, Monoclonal/immunology , Antibodies, Neoplasm/immunology , Antigens, Differentiation/immunology , Antigens, Surface/immunology , Antigens, Tumor-Associated, Carbohydrate/chemistry , B-Lymphocytes/cytology , Cattle , Cell Differentiation , Enzootic Bovine Leukosis/microbiology , Flow Cytometry , Molecular Weight
18.
Can J Vet Res ; 56(4): 339-48, 1992 Oct.
Article in English | MEDLINE | ID: mdl-1335834

ABSTRACT

Twenty-seven cattle with lymphoma and 46 cows from a known bovine leukemia virus (BLV)-infected herd were tested for anti-BLV antibody by the agar gel immunodiffusion (AGID) test and an enzyme-linked immunosorbent assay (ELISA). The polymerase chain reaction (PCR) and Southern hybridization were used to detect BLV provirus in the tumor DNA of the 27 cattle with lymphoma. The PCR was used to detect BLV provirus in the peripheral blood mononuclear cell DNA of the 46 normal known-exposed cattle. Two presumed false negative AGID test results compared to ELISA were found. Of ten cattle three years of age or less with "sporadic" forms of lymphoma, four had BLV provirus in tumor DNA, detectable by PCR. In two of these four, BLV provirus was clonally integrated based on digestion of tumor DNA with restriction enzymes followed by Southern hybridization. The BLV provirus was not detected by PCR in 5 of 17 cattle with "enzootic" lymphoma and two of these five were seronegative. Among normal BLV-exposed cows, 6.5% (3 of 46) were serologically positive and PCR negative; serologically negative and PCR positive cows occurred with the same frequency. Serological and PCR test results, when considered in all cattle (n = 73), had a concordance rate of 83.6%. Discordant test results occurred with approximately equal frequency between serologically positive and PCR negative (7 of 73, 9.6%) and serologically negative and PCR positive (5 of 73, 6.8%) groups. These data suggest that the role of BLV in some "sporadic" bovine lymphomas, previously unassociated with BLV, should be reexamined. The BLV provirus was not demonstrable in the tumor DNA from five adult cattle with lymphoma, suggesting that BLV may not be the etiological agent in all adult bovine lymphomas. The findings of persistently seronegative PCR positive and seropositive PCR negative cattle indicate that further work is needed to more fully understand the host-virus interaction. Present serological screening methods may not have sufficient sensitivity for determining BLV status in some circumstances.


Subject(s)
Enzootic Bovine Leukosis/microbiology , Leukemia Virus, Bovine/isolation & purification , Lymphoma/veterinary , Proviruses/isolation & purification , Animals , Antibodies, Viral/blood , Base Sequence , Blotting, Southern , Cattle , DNA, Viral/analysis , DNA, Viral/chemistry , Enzyme-Linked Immunosorbent Assay , Female , Immunodiffusion , Leukemia Virus, Bovine/genetics , Leukemia Virus, Bovine/immunology , Lymphoma/microbiology , Male , Molecular Sequence Data , Nucleic Acid Hybridization , Polymerase Chain Reaction , Proviruses/genetics , Proviruses/immunology
19.
Dtsch Tierarztl Wochenschr ; 99(5): 191-3, 1992 May.
Article in German | MEDLINE | ID: mdl-1322267

ABSTRACT

Comprehensive serological and virological monitoring for bovine viral diarrhoea (BVD) virus was applied in a dairy herd. Out of 83 calves 26 persistently infected animals were identified. Four viremic calves showed clinical signs of disease, the others displayed no symptoms. Viral isolates from persistently infected animals were homogenous with respect to their antigenicity. The results of virological and serological investigations allowed an almost complete reconstruction of events following the introduction of BVD virus into the herd. This case illustrates the potentially dangerous and damaging effects of unidentified virus carriers in cattle herds. Strategies for the identification of virus-shedding animals and the limitation of economical losses are discussed.


Subject(s)
Carrier State/veterinary , Diarrhea Viruses, Bovine Viral/immunology , Enzootic Bovine Leukosis/microbiology , Viremia/veterinary , Animals , Antibodies, Viral/blood , Antigens, Viral/blood , Carrier State/microbiology , Cattle , Viremia/microbiology
20.
Microbiol Immunol ; 36(12): 1317-23, 1992.
Article in English | MEDLINE | ID: mdl-1337572

ABSTRACT

Lymphocyte proliferation responses were investigated in sheep and cattle, in which the replication of bovine leukemia virus (BLV) had been known to be suppressed by inoculation with recombinant vaccinia virus (rVV) expressing BLV envelope glycoprotein (gp60). Enhanced lymphocyte proliferation responses were observed in animals inoculated with rVV, regardless of whether they were naive or BLV carriers. These responses were roughly inversely correlated to the growth of BLV in the peripheral blood leukocytes. In contrast, there was no apparent correlation between humoral immune response and BLV growth. Based on these results, it was suggested that rVV rendered its suppressive effect of BLV replication primarily via augmentation of cell-mediated immunity.


Subject(s)
Enzootic Bovine Leukosis/immunology , Leukemia Virus, Bovine/immunology , Lymphocyte Activation , Viral Envelope Proteins/immunology , Viral Vaccines/immunology , Animals , Cattle , Enzootic Bovine Leukosis/microbiology , Leukemia Virus, Bovine/physiology , Leukemia, Experimental/immunology , Leukemia, Experimental/microbiology , Leukocytes/microbiology , Male , Sheep , Sheep Diseases/immunology , Sheep Diseases/microbiology , Vaccination , Vaccines, Synthetic/immunology , Vaccinia virus , Virus Replication
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