Determination of proviral load in bovine leukemia virus–infected cattle with and without lymphocytosis

Marcela A. Juliarena Laboratorio de Virologia, Departamento de Sanidad Animal y Medicina Preventiva, Facultad de Veterinaria, Universidad Nacional del Centro de la Provincia de Buenos Aires, Pinto 399, Tandil, Argentina.

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Silvina E. Gutierrez Laboratorio de Virologia, Departamento de Sanidad Animal y Medicina Preventiva, Facultad de Veterinaria, Universidad Nacional del Centro de la Provincia de Buenos Aires, Pinto 399, Tandil, Argentina.

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Carolina Ceriani Departamento de Ciencias Biologicas, Facultad de Veterinaria, Universidad Nacional del Centro de la Provincia de Buenos Aires, Pinto 399, Tandil, Argentina.

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Abstract

Objective—To determine proviral load in bovine leukemia virus (BLV)–infected cattle with and without persistent lymphocytosis to assess the potential of transmitting the virus.

Animals—Cattle in 6 dairy herds.

Procedures—Blood samples from infected cows were evaluated 3 times at 6-month intervals for determination of proviral load via PCR assay, serologic results via ELISA, and hematologic status via differential cell counts.

Results—Infected cattle were classified into lymphocytotic and nonlymphocytotic groups. Lymphocytotic cattle consistently had > 100,000 copies of integrated provirus/μg of DNA (ie, high proviral load) in peripheral blood leukocytes. Titers of antibodies against BLVgp51 and BLVp24 indicated a strong immune response. Nonlymphocytotic cattle comprised 2 subgroups: a group with high proviral load and strong immune response, and a group with a weaker immune response, mostly against BLVp24, and a proviral load of < 100 copies/μg of DNA (ie, low proviral load).

Conclusions and Clinical Relevance—Results emphasized the importance of characterizing nonlymphocytotic BLV-infected cattle during eradication programs. The risk of transmitting BLV infection from nonlymphocytotic cattle may differ depending on the proviral load. Nonlymphocytotic cattle with high proviral load could be efficient transmitters (as efficient as lymphocytotic cattle), whereas nonlymphocytotic cattle with low proviral load could be inefficient transmitters under standard husbandry conditions. Because most cattle with low proviral load do not develop anti-BLVp24 antibodies, it appears that lack of an anti-BLVp24 antibody response may be a good marker of this condition.

Abstract

Objective—To determine proviral load in bovine leukemia virus (BLV)–infected cattle with and without persistent lymphocytosis to assess the potential of transmitting the virus.

Animals—Cattle in 6 dairy herds.

Procedures—Blood samples from infected cows were evaluated 3 times at 6-month intervals for determination of proviral load via PCR assay, serologic results via ELISA, and hematologic status via differential cell counts.

Results—Infected cattle were classified into lymphocytotic and nonlymphocytotic groups. Lymphocytotic cattle consistently had > 100,000 copies of integrated provirus/μg of DNA (ie, high proviral load) in peripheral blood leukocytes. Titers of antibodies against BLVgp51 and BLVp24 indicated a strong immune response. Nonlymphocytotic cattle comprised 2 subgroups: a group with high proviral load and strong immune response, and a group with a weaker immune response, mostly against BLVp24, and a proviral load of < 100 copies/μg of DNA (ie, low proviral load).

Conclusions and Clinical Relevance—Results emphasized the importance of characterizing nonlymphocytotic BLV-infected cattle during eradication programs. The risk of transmitting BLV infection from nonlymphocytotic cattle may differ depending on the proviral load. Nonlymphocytotic cattle with high proviral load could be efficient transmitters (as efficient as lymphocytotic cattle), whereas nonlymphocytotic cattle with low proviral load could be inefficient transmitters under standard husbandry conditions. Because most cattle with low proviral load do not develop anti-BLVp24 antibodies, it appears that lack of an anti-BLVp24 antibody response may be a good marker of this condition.

Contributor Notes

Supported in part by Agencia Nacional de Promocion Cientifica y Tecnologica (PICT 2002; Proyecto BID 1728 OC/AR No. 08/11083), Consejo Nacional de Investigaciones Cientificas y Tecnicas (Res. No. 650/04), and SECAT (UNCPBA).

Presented in part at the VIII Congress of Virology, Buenos Aires, September 2005.

The authors thank Patricia Bani and Norma Rodriguez for technical assistance.

Address correspondence to Dr. Ceriani.
  • 1.

    Sprecher DJ, Pelzer KD, Lessard P. Possible effect of altered management practices on seroprevalence of bovine leukemia virus in heifers of a dairy herd with history of high prevalence of infection. J Am Vet Med Assoc 1991;199:584588.

    • Search Google Scholar
    • Export Citation
  • 2.

    Mahieux R, Gessain A. New human retroviruses: HTLV-3 and HTLV-4. Med Trop 2005;65:525528.

  • 3.

    Rosenberg N, Jolicoeur P. Retroviral pathogenesis. In: Coffin JM, Hughes SH, Varmus HE, eds. Retrovirology. Woodbury, NY: Cold Spring Harbor Laboratory Press, 1997;475586.

    • Search Google Scholar
    • Export Citation
  • 4.

    Uchiyama T. Human T cell leukemia virus type I (HTLV-I) and human diseases. Annu Rev Immunol 1997;15:1537.

  • 5.

    Burny A, Bruck C, Cleuter Y, et al. Bovine leukaemia virus and enzootic bovine leukosis. Onderstepoort J Vet Res 1985;52:133144.

  • 6.

    Ferrer JF. Bovine lymphosarcoma. Adv Vet Sci Comp Med 1980;24:168.

  • 7.

    Debacq C, Asquith B, Reichert M, et al. Reduced cell turnover in bovine leukemia virus-infected, persistently lymphocytotic cattle. J Virol 2003;77:1307313083.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 8.

    Burny A, Bruck C, Chantrenne H, et al. Bovine leukaemia virus: biology and epidemiology. In: Klein G, ed. Viral oncology. New York: Raven Press, 1980;231289.

    • Search Google Scholar
    • Export Citation
  • 9.

    Suh GH, Lee JC, Lee CY, et al. Establishment of a bovine leukaemia virus-free dairy herd in Korea. J Vet Sci 2005;6:227230.

  • 10.

    Reichel MP, Tham KM, Barnes S, et al. Evaluation of alternative methods for the detection of bovine leukaemia virus in cattle. N Z Vet J 1998;46:140146.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11.

    Simard C, Richardson S, Dixon P, et al. Enzyme-linked immunosorbent assay for the diagnosis of bovine leucosis: comparison with the agar gel immunodiffusion test approved by the Canadian Food Inspection Agency. Can J Vet Res 2000;64:101106.

    • Search Google Scholar
    • Export Citation
  • 12.

    Fechner H, Burg A, Geue L, et al. Evaluation of polymerase chain reaction (PCR) application in diagnosis of bovine leukaemia virus (BLV) infection in naturally infected cattle. Zentralbl Veterinarmed [B] 1996;43:621630.

    • Search Google Scholar
    • Export Citation
  • 13.

    Murtaugh MP, Lin GF, Haggard DL, et al. Detection of bovine leukaemia virus in cattle by the polymerase chain reaction. J Virol Methods 1991;33:7385.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 14.

    Alejo D, Gutiérrez SE, Dolcini GL, et al. Prevalencia de la infección por el virus de la leucosis bovina (BLV) en tambos de los partidos de General Pueyrredón y Balcarce. Rev Asoc Argent Produccion Anim 2000;20:7783.

    • Search Google Scholar
    • Export Citation
  • 15.

    Trono KG, Perez-Filgueira DM, Duffy S, et al. Seroprevalence of bovine leukemia virus in dairy cattle in Argentina: comparison of sensitivity and specificity of different detection methods. Vet Microbiol 2001;83:235248.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 16.

    Gutierrez SE, Dolcini GL, Arroyo GH, et al. Development and evaluation of a highly sensitive and specific blocking enzymelinked immunosorbent assay and polymerase chain reaction assay for diagnosis of bovine leukemia virus infection in cattle. Am J Vet Res 2001;62:15711577.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 17.

    Esteban EN, Thorn R, Ferrer JF. Characterization of the blood lymphocyte population in cattle infected with the bovine leukaemia virus. Cancer Res 1985;45:32253230.

    • Search Google Scholar
    • Export Citation
  • 18.

    Mirsky ML, Olmstead C, Da Y, et al. Reduced bovine leukaemia virus proviral load in genetically resistant cattle. Anim Genet 1998;29:245252.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 19.

    Abt DA, Marshak RR, Kulp HW, et al. Studies on the relationship between lymphocytosis and bovine leucosis. Bibl Haematol 1970;36:527536.

    • Search Google Scholar
    • Export Citation
  • 20.

    Stear MJ, Dimmock CK, Newman MJ, et al. BoLA antigens are associated with increased frequency of persistent lymphocytosis in bovine leukaemia virus infected cattle and with increased incidence of antibodies to bovine leukaemia virus. Anim Genet 1988;19:151158.

    • Search Google Scholar
    • Export Citation
  • 21.

    Gutierrez SE, Dolcini GL, Bani PC, et al. Respuesta immune humoral contra el virus de la leucosis bovina (BLV) en bovinos con distintos niveles de carga viral (CV), linfocitosis (LP) y linfosarcoma (LFS). Medicina 1997;57:3435.

    • Search Google Scholar
    • Export Citation
  • 22.

    Bendixen HJ. Bovine enzootic leukosis. Adv Vet Sci Comp Med 1965;10:129204.

  • 23.

    Theilen GH, Madewell BR. Hematopoietic neoplasms, sarcomas and related conditions. Part V: bovine. In: Theilen GH, Madewell BR, eds. Veterinary cancer medicine. 2nd ed. Philadelphia: Lea & Febiger, 1987;408432.

    • Search Google Scholar
    • Export Citation
  • 24.

    Madisen L, Hoar DI, Holroyd CD, et al. DNA banking: the effects of storage of blood and isolated DNA on the integrity of DNA. Am J Med Genet 1987;27:379390.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 25.

    Sagata N, Yasunaga T, Tsuzuku-Kawamura J, et al. Complete nucleotide sequence of the genome of bovine leukaemia virus: its evolutionary relationship to other retroviruses. Proc Natl Acad Sci U S A 1985;82:677681.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 26.

    Van den Broeke A, Cleuter Y, Beskorwayne T, et al. CD154 costimulated ovine primary B cells, a cell culture system that supports productive infection by bovine leukemia virus. J Virol 2001;75:10951103.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 27.

    Juliarena MA, Gutierrez SE, Ceriani C. Chicken antibodies: a useful tool for antigen capture ELISA to detect bovine leukemia virus without cross-reaction with other mammalian antibodies. Vet Res Commun 2007;31:4351.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 28.

    Greiner M, Sohr D, Göbel P. A modified ROC analysis for the selection of cut-off values and the definition of intermediate results of serodiagnostic tests. J Immunol Methods 1995;185:123132.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 29.

    Greiner M. Two-graph receiver operating characteristic (TG-ROC): a Microsoft-EXCEL template for the selection of cut-off values in diagnostic tests. J Immunol Methods 1995;185:145146.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 30.

    Cockerell GL, Rovnak J. The correlation between the direct and indirect detection of bovine leukemia virus infection in cattle. Leuk Res 1988;12:465469.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 31.

    Murtaugh MP, Lin GF, Haggard DL, et al. Detection of bovine leukemia virus in cattle by the polymerase chain reaction. J Virol Methods 1991;33:7385.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 32.

    Eaves FW, Molloy JB, Dimmock CK, et al. A field evaluation of the polymerase chain reaction procedure for the detection of bovine leukaemia virus proviral DNA in cattle. Vet Microbiol 1994;39:313321.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 33.

    Burny A, Cleuter Y, Kettmann R, et al. Bovine leukaemia: facts and hypotheses derived from the study of an infectious cancer. Cancer Surv 1987;6:139159.

    • Search Google Scholar
    • Export Citation
  • 34.

    Molloy JB, Dimmock CK, Eaves FW, et al. Control of bovine leukaemia virus transmission by selective culling of infected cattle on the basis of viral antigen expression in lymphocyte cultures. Vet Microbiol 1994;39:323333.

    • Crossref
    • Search Google Scholar
    • Export Citation

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