Evaluation of a canine transmissible venereal tumour cell line with tumour immunity capacity but without tumorigenic property

Open access


Introduction: Canine transmissible venereal tumour (CTVT) is a sexually transmitted tumour affecting dogs worldwide, imposing a financial burden on dog owners. A stable culture cell line in continuous passages for >18 months has only been achieved once. The present study investigated a stable CTVT cell line isolated from a bitch and its potential as a vaccine.

Material and Methods: A biopsy from a 2-year-old mongrel bitch with CTVT was obtained for histopathological confirmation and isolation of tumour cells. The isolated cells were cultured to passage 55 and characterised by flow cytometry, with karyotyping by GTG-banding and by PCR detection of myc S-2 and LINE AS1. The isolated CTVT cell line was also used as a preventive vaccine in a canine model.

Results: Histopathological analysis of the isolated tumour cells revealed typical CTVT characteristics. Constant proliferation and stable morphological characteristics were observed during culture. Phenotypic analysis determined the expression of HLA-DR+, CD5.1+, CD14+, CD45+, CD83+, CD163+, and Ly-6G-Ly-6C+. GTG-banding revealed a mean of 57 chromosomes in the karyotype with several complex chromosomal rearrangements. LINE-c-myc insertion in the isolated CTVT cell line at 550 bp was not detected. However, a 340-bp band was amplified. Isolated CTVT cell line inoculation at a concentration of 1×108 did not induce tumour growth in bitches, nor did a challenge with primary CTVT cells.

Conclusion: The present study successfully identified and isolated a stable CTVT cell line that may be useful in CTVT prevention.

1. Adams E.W., Carter L.P., Sapp W.J.: Growth and maintenance of the canine venereal tumor in continuous culture. Cancer Res 1968, 28, 753–757.

2. Antonov A.: Successful treatment of canine transmissible venereal tumor using vincristine sulfate. Adv Res 2015, 5, 1–5.

3. Bloom F., Paff G.H., Nobace C.R.: The transmissible venereal tumor of the dog. Studies indicating that the tumor cells are mature end cells of reticulo-endothelial origin. Am J Pathol 1950, 27, 119–139.

4. Ching Y.L., His H.L., Ming J.T., Wang Y.K.: Vimentin contributes to epithelial-mesenchymal transition cancer cell mechanics by mediating cytoskeletal organization and focal adhesion maturation. Oncotarget 2015, 6, 15966–15983.

5. Conn M.P.: Sourcebook of models for biomedical research. Springer, Oregon Health and Science University, Portland 2008, pp. 145–158.

6. Das U., Das A.K.: Review of canine transmissible venereal sarcoma. Vet Res Commun 2000, 24, 545–556.

7. De La Sota P., Amigo G., Adagio L., Noia M., Gobello C.: Transmissible venereal tumor in the dog. Vet Sci 2000, 4, 1515–1883.

8. Ferreira D., Adega F., Chaves R.: The importance of cancer cell lines as in vitro models in cancer methylome analysis and anticancer drugs testing. INTECH, Institute for Biotechnology, Portugal 2013, pp. 139–166.

9. Flórez M.M., Ballestero H.F., Duzanski A.P., Bersano R.O., Lima J.F., Cruz F.L., Mota L.S., Rocha N.S.: Immunocytochemical characterization of primary cell culture in canine transmissible venereal tumor. Pesq Vet Bras 2016, 36, 844–850.

10. Fonseca L.S., Mota S.L., Colodel M.M., Ferreira I., Brandão C.V., Rocha N.S.: Spontaneous canine transmissible venereal tumor: association between different phenotypes and the insertion LINE-1/c-myc. Rev Colombiana de Ciencias Pecuarias 2012, 25, 402–408.

11. Fridman W.H., Pagés F., Sautés C., Galon J.: The immune contexture in human tumours: impact on clinical outcome. Nat Rev Cancer 2012, 12, 298–306.

12. Girma B., Mersha C.: A review on canine transmissible venereal tumor: from morphologic to biochemical and molecular diagnosis. Acad J Anim Dis 2015, 4, 185–195.

13. Kollaritsch H., Wagner P.R.: Travel Medicine. Live vaccines. Trop Dis Travel Med Vaccines 2013, 67–76.

14. Legare M.E., Bush J., Ashley A.K., Kato T., Hanneman W.H.: Cellular and phenotypic characterization of canine osteosarcoma cell lines. J Cancer 2011, 2, 262–270.

15. Liao K.W., Lin Z.Y., Pao H.N., Kam S.Y., Wang F.I., Chu R.M.: Identification of canine transmissible venereal tumor cells using in situ polymerase chain reaction and the stable sequence of the long interspersed nuclear element. J Vet Diagn Invest 2003, 15, 399–406.

16. McCallum H.: Tasmanian devil facial tumour disease: lessons for conservation biology. Trends Ecol Evol 2008, 11, 631–637.

17. Mukaratirwa S., Gruys E.: Canine transmissible venereal tumour: cytogenetic origin, immunophenotype, and immunobiology. Vet Quart 2003, 25, 101–111.

18. Murchison E.P.: Clonally transmissible cancers in dogs and Tasmanian devils. Oncogene 2009, 27, 19–30.

19. Murchison E.P., Wedge D.C., Alexandrov L.B., Beiyuan F., Martincorena I., Ning Z., Tubio J.M., Werner E.I., Allen J., De Nardi A.B., Donelan E.M., Marino G., Fassati A., Campbell P.J., Yang F., Burt A., Weiss R.A., Stratton M.R.: Transmissible dog cancer genome reveals the origin and history of an ancient cell lineage. Science 2014, 343, 437–440.

20. Murgia C., Pritchard J.K., Kim S.Y., Fassati A., Weiss R.A.: Clonal origin and evolution of a transmissible cancer. Cell 2006, 7, 477–487.

21. Nowinsky M.A.: Zur Frage über die Impfung der Krebsigen Geschwülste. Zentr Bl. Med.Wissensch 1876, 14, 790-791.

22. O’Neill I.D.: Concise review: transmissible animal tumors as models of the cancer stem-cell process. Stem Cells Dayt Ohio 2011, 29, 1909–1914.

23. Ostrander E.A., Davis B.W., Ostrander G.K.: Transmissible tumors: breaking the cancer paradigm. Trends in Genetics 2016, 32, 1–15.

24. Pai C.C., Kuo T.F., Mao S.J.T., Chuang T.F., Lin C.S., Chu R.M.: Immunopathogenic behaviors of canine transmissible venereal tumor in dogs following an immunotherapy using dendritic/tumor cell hybrid. Vet Immunol Immunopathol 2011, 139, 187–199.

25. Prier J.E.: Nutritional requirements for cultivation of the canine transmissible sarcoma cells. Cancer Res 1962, 3, 695–699.

26. Purohit G.: Canine transmissible venereal tumor: a review. Internet J Vet Med 2008, 6, 1–13.

27. Ramírez R.R., García J.S., Nevarez G., Rodríguez T.L.: Transmissible venereal tumor with metastasis to a spleen hemangioma in a bitch. Vet Méx 2010, 2, 21–30.

28. Siddle H.V., Kaufman J.: Immunology of naturally transmissible tumours. Immunology 2015, 144, 11–20.

29. Stein G.S.: ATCC primary cell culture guide, tips and techniques for culturing primary cells. American Type Culture Collection, Manassas 2012, pp. 34–45.

30. Stockmann D., Ferrari H.F., Andrade A.L., Lopes R.A., Cardoso T.C., Luvizotto M.C.R.: Canine transmissible venereal tumors: aspects related to programmed cell death. Braz J Vet Pathol 2011, 4, 67–75.

31. Strakova A., Murchison E.P.: The changing global distribution and prevalence of canine transmissible venereal tumour. BMC Vet Res 2014, 10, 168–173.

32. Thalhammer S., Koehler U., Stark R.W., Heckl W.M.: GTG banding pattern on human metaphase chromosomes revealed by high resolution atomic-force microscopy. J Microsc 2001, 202, 464–467.

Journal of Veterinary Research

formerly Bulletin of the Veterinary Institute in Pulawy

Journal Information

IMPACT FACTOR J Vet Res 2017: 0.811

CiteScore 2017: 0.68

SCImago Journal Rank (SJR) 2017: 0.29
Source Normalized Impact per Paper (SNIP) 2017: 0.484


All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 39 39 17
PDF Downloads 24 24 11