Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
권호 표지

Cultivation and Electron Microscopy of Bovine Leukemia Virus from Peripheral Blood Lymphocytes of Holstein-Friesian Dairy Cattle

젖소 말초혈액 림프구로부터 소백혈병 바이러스 배양 및 전자현미경적 관찰

  • Yoon, Soon-Seek (National Veterinary Research and Qurantine Service) ;
  • Park, Jung-Won (National Veterinary Research and Qurantine Service) ;
  • Byun, Jae-Won (National Veterinary Research and Qurantine Service) ;
  • Kang, Mun-Il (College of Veterinary Medicine, Chonnam National University) ;
  • Yoo, Han-Sang (College of Veterinary Medicine, Seoul National University) ;
  • Han, Hong-Ryul (College of Veterinary Medicine, Seoul National University)
  • Published : 2005.03.01
Download PDF
⟨ Previous Next ⟩

Abstract

Many studies have been performed on the bovine leukemia virus (BLV) since bovine leukosis had been reported in 1968 in Korea. However, there was no report on the ultrastructural examination of BLV. An attempt to detect C-type viral particles in the cultured peripheral blood lymphocytes of Holstein-Friesian dairy cattle, was made to determine whether in vitro viral expression might be used as a reliable method to identify the cow which is likely to transmit BLV. In transmission electron microscopic (TEM) examination, the virus particles were found predominantly outside of the lymphocytes even though a few particles were also observed within the membrane bound cytoplasmic vacuoles. All of them were C-type particles consisting of a central, electron-dense core separated by a clear area from a limiting envelope with a unit membrane structure. Virus particles were easily detected in the lymphocyte which was cultured with medium supplemented with either T-lymphocyte mitogen (conconavalin A) or B-lymphocyte mitogen (lipopolysaccharide). Identical viral particles, although fewer, were also consistently present in the lymphocytes cultured with medium which was containing foetal bovine serum (FBS) only and which was containing neither FBS or mitogen. By contrast, no virus particle was detected in extensive examination of lymphocytes before culture. In conclusion, the BLV cultivation and detection methods established in this study could be used as a tool to identify and eliminate the cattle which can transmit the BLV.

국내 젖소의 54.2%가 BLV에 감염되어 있지만 현재까지 국내에서는 소백혈병 바이러스 (BLV) 입자를 확인한 연구 보고가 없기 때문에 BLV 항체 양성 소의 말초혈액 림프구를 배양, BLV를 발현시켜 전자현미경으로 바이러스 입자를 검출하였고 배양조건에 따른 바이러스 발현율 및 발현 시간을 비교하였다. 검사 결과 전형적인 C-형 바이러스를 확인할 수 있었으며 BLV 단크론 항체를 이용한 면역염색결과 BLV 항원 양성으로 확인되었다. BLV는 대부분 세포 외부에 분포하고 있었으며 세포질 막에서 생성, 발아되어 나오는 것도 관찰되었다. 전체 바이러스의 크기는 $90{\sim}100$ nm였으며 nucleocapsid는 $40{\sim}60$nm였다. 소태아혈청 (FBS)과 T- 및 B-림프구 분열촉진물질(mitogen)을 각각 첨가하여 배양한 결과 두 군 모두에서 BLV 발현이 확인되었다. Lipopolysaccharide 첨가군은 배양 12시간, Conconavalin A 첨가군은 배양 24시간에 각각 림프구의 10%에서 바이러스가 관찰되었다. 또한 FBS만 첨가한 군과 FBS와 mitogen을 모두 첨가하지 않은 군에서도 관찰되었으나 바이러스의 수는 적었다. 본 연구에서 확립된 BLV 배양 기법을 활용하면 BLV에 감염된 소 중 바이러스를 발현하는 소, 즉 전파능이 있는 개체를 찾아내어 우선적으로 도태할 수 있기 때문에 BLV 감염으로 인한 피해를 막는데 효과적으로 이용될 수 있을 것으로 사료된다.

Keywords

References

  1. Baliga V, Ferrer JF: Expression of the bovine leukemia virus and its internal antigen in blood lymphocytes. Proc Soc Exp Biol Med 156 : 388-391, 1977
  2. Ballagi-Pordany A, Belak S: The use of mimics as internal standards to avoid false negatives in diagnostic PCR. Molecular and Cellular Probes 10 : 159-164, 1996 https://doi.org/10.1006/mcpr.1996.0022
  3. Bendixen HJ: Bovine Enzootic Leukosis. Advance in Vet Sci and Comp Med 10 : 129-204, 1965
  4. Burny A, Bex F, Chantrenne H, Cleute Y, Dekegel D, Ghysdael J, Kettmann R, Leclercq M, Leunen J, Mammerickx M, Portetelle D: Bovine leukemia virus involvement in enzootic bovine leukosis. Advance Can Res 28 : 251-311, 1978 https://doi.org/10.1016/S0065-230X(08)60649-1
  5. Burny A, Mammerickx M: Enzootic bovine leukosis and bovine leukemia virus. Kluwer. Martinus Nijhoff Publishing. Boston, 1987
  6. Chevallier N, Berthelemy M, Rhun DL, Laine V, Levy D, Cornil IS: Bovine leukemia virus-induced lymphocytosis and increased cell survival mainly involve the $CD11b^{+}$ B-lymphocyte subset in sheep. J Virol 72(5) : 4413-4420, 1998
  7. Choi YB, Kim YH, Lee JH, Kim JS: Effects of squalene on the epidermal growth factor (EGF) expression and histological changes by glycerol-induced acute renal failure in mice. Kor J Electron Microscopy 34(4) : 241-254, 2004. (Korean)
  8. Dequiedt F, Cantor GH, Hamilton VT, Pritchard SM, Davis WC, Kerkhofs P, Burny A, Kettmann R, Willems L: Bovine leukemia virus-induced persistent lymphocytosis in cattle does not correlate with increased Ex Vivo survival of lymphocytes. J Virol 73(2) : 1127-1137, 1999
  9. Fields BN, Knipe DM: Virology. Second edition. Vol 2: pp 1437-1500. Raven press, New York, 1990
  10. Gupta P, Ferrer JF: Expression of bovine leukemia virus genome is blocked by a nonimmunoglobulin protein in plasma from infected cattle. Science 215 : 405-406, 1982 https://doi.org/10.1126/science.6276975
  11. Gupta P, Kashmiri SVS, Ferrer JF: Transcriptional control of the bovine leukemia virus genome: Role and characterization of a non-immunoglobulin plasma protein from bovine leukemia virus-infected cattle. J Virol 50(1) : 267-270, 1984
  12. Johnson R, Kaneene JB: Bovine leukemia virus. Part IV. Economic impact and control measures. Compendium 13(11) : 1727-1737, 1991
  13. Jun MH, Chung UI, Lee CK, Baig SY, Lim CH: Seroepizootiological study on bovine leukosis in Korea. Korean J Vet Res 22(2) : 175-185, 1982
  14. Keefe RG, Choi Y, Ferrick DA, Stott JL: Bovine cytokine expression during different phases of bovine leukemia virus infection. Vet Immunol Immunopathol 56 : 39-51, 1997 https://doi.org/10.1016/S0165-2427(96)05727-3
  15. Maruyama K, Fukushima T, Mochizuki S: Cross-reactive antibodies to BLV and HTLV in bovine and human hosts with retrovirus infection. Vet Immunol Immunpathol 22 : 265-274, 1989 https://doi.org/10.1016/0165-2427(89)90013-5
  16. Miller JM, Miller LD, Olson C, Gillette KG: Virus-like particle in phytohemagglutinin-stimulated lymphocyte cultures with reference to bovine lymphosarcoma. J Natl Cancer Inst 43 : 1297-1305, 1969
  17. Miller LD, Miller JM, Maaten MJ, Schmerr MJF: Blood from bovine leukemia virus-infected cattle: Antigen production correlated with infectivity. Am J Vet Res 46(4) : 808-810, 1985
  18. Miller LD, Miller JM, Olson C: Inoculation of calves with particles resembling C-type virus from cultures of bovine lymphosarcoma. J Natl Cancer Inst 48 : 423-428, 1972
  19. Murphy FA, Fauquet CM, Bishop DHL, Ghabriel SA, Janis AW, Martelli GP, Mayo MA, Summers MD: Virus Taxonomy (Classification and nomenclature of viruses). Sixth report of the international committee on taxonomy of viruses. pp. 193-204. Springer-Verlag Wien, New York, 1995
  20. Ohshima K, Miura S, Numakunai S, Sano H, Suganuma T: C-type virus particles associated with Phytohemagglutinin-stimulated buffy-coat-cells of leukemic cows. Jpn J Vet Sci 39 : 77-82, 1977 https://doi.org/10.1292/jvms1939.39.77
  21. Olson C, Miller JM, Miller LD, Gillette KG: C-type virus and lymphocytic nuclear projections in bovine lymphosarcoma. JAVMA 156(12) : 1880-1883, 1970
  22. Olson C: Bovine lymphosarcoma (leukemia). JAVMA 165 : 630-632, 1974
  23. Paul PS, Pomeroy KA, Johnson DW, Muscoplat CC, Handwerger BS, Soper FF, Sorensen DK: Evidence for the replication of bovine leukemia virus in the B lymphocytes. Am J Vet Res 38(6) : 873-876, 1977
  24. Pyeon D, O'reilly KL, Splitter GA: Increased interleukin-10 mRNA expression in tumor-bearing or persistently lymphocytotic animals infected with bovine leukemia virus. J Virol 70(8) : 5706-5710, 1996
  25. Rebhun WC, Guard C, Richard CM: Diseases of Dairy Cattle. pp. 484-491. William and Wilkins, Baltimore, 1995
  26. Schwartz I, Levy D: Pathobiology of bovine leukemia virus. Vet Res 25 : 521-536, 1994
  27. Sordillo LM, Hicks CR, Pighetti GM: Altered interleukin-2 production by lymphocyte populations from bovine leukemia virus-infected cattle. Proc Soc Exp Biol Med 207 : 268-273, 1994
  28. Stock ND, Ferrer J: Replicating C-type virus in phytohemaglutinin-treated buffy-coat cultures of bovine origin. J Natl Cancer Inst 48 : 985-996, 1972
  29. Suh GH: Establishment of a bovine leukemia virus-free dairy cattle. Ph.D. Thesis, Cheonnam national university, Kwangju, South Korea, 2004. (Korean)
  30. Trueblood ES, Crown WC, Palmer GH, Davis WC, Stone DM, McElwain TF: B-lymphocyte proliferation during bovine leukemia virus-induced persistent lymphocytosis is enhanced by T-lymphocyte-derived interleukin-2. J Virol 72(4) : 3169-3177, 1998
  31. Weber A, M. Fahning, Hammer RH, Jessen C: Relationship between nuclear pockets in bovine peripheral blood lymphocytes and C-type virus particles in cultures of these cells. J Natl Cancer Inst 51 : 81-88, 1973 https://doi.org/10.1093/jnci/51.1.81
  32. Weber AF, Meiske JC, Hooker EC, Haggard DL, Domagala AM, Sorensen DK, Buoen LC: In vitro viral expression as a criterion for development of control procedures for enzootic bovine leukosis. Am J Vet Res 48(6) : 899-903, 1987
  33. Yoon SS, Bae YC, Lee KH, Han B, Han HR: Characteristics of bovine lymphoma caused by bovine leukemia virus infection in Holstein-Friesian dairy cattle in Korea. Asian-Aust J Anim Sci 18(5) : 728-733, 2005 https://doi.org/10.5713/ajas.2005.728