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Diagnostic implications of the molecular forms and levels of serum thymidine kinase 1 in different canine malignancies

Jagarlamudi, Kiran Kumar (2015). Diagnostic implications of the molecular forms and levels of serum thymidine kinase 1 in different canine malignancies. Diss. (sammanfattning/summary) Uppsala : Sveriges lantbruksuniv., Acta Universitatis agriculturae Sueciae, 1652-6880 ; 2015:32
ISBN 978-91-576-8262-8
eISBN 978-91-576-8263-5
[Doctoral thesis]

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Abstract

Thymidine kinase 1 (ATP: thymidine 5’- phosphotransferase, EC 2.1.7. 21, TK1) is a cellular enzyme involved in salvage pathway for DNA precursor synthesis. TK1 activity fluctuates during cell cycle, it reaches peak in S phase and absent in M phase. Because of its tight regulation with cell cycle, TK1 has been used as proliferation marker for diagnosis and treatment monitoring of various malignancies in human and veterinary medicine. TK1 levels can be measured by activity based or antibody based assays.

The main aim of the research described in this thesis was to develop TK1 antibody based assays for determining serum TK1 protein levels in comparison with TK1 activity from dogs with different malignancies. Further analysis revealed a significant difference in the molecular forms of TK1 in sera from canine leukemia and mammary tumours.

In Study I, serum TK1 protein levels in dogs with
different solid tumours were determined by using an antibody based assay i.e. immunoaffinity assay. TK1 protein levels were significantly higher in dogs with solid tumours than expected from the TK1 activity values. In contrast, the specific activity of TK1 in sera from healthy dogs was 2.5 fold higher than that of solid tumours. The molecular forms of recombinant, cellular and serum TK1 were investigated in study II. Dog recombinant and serum TK1 existed as oligomers with a molecular weight (MW) of 720-300 kDa. Cellular TK1 from both dogs and humans were mainly as tetramers. Human recombinant and serum TK1 activity eluted in two peaks: one at high and one at low MW, corresponding to 720-300 kDa and 200-50 kDa, respectively.

In study III, TK1 protein levels in sera from dogs with mammary tumours were determined by immunoaffinity assay. The TK1 protein assay differentiated mammary adenomas efficiently from healthy dogs, and adenomas from carcinomas, but this was not possible with the TK1 activity assay. In mammary tumour sera, active TK1 eluted as high MW oligomer similar to leukemia however, TK1 protein was detected not only as high MW form but also in the fractions where no TK1 activity was found. This indicates that serum TK1 exits in multiple forms in mammary tumours with a large fraction of inactive TK1 protein. Study IV describes the development of TK1-ELISA, based on a polyclonal and a monoclonal anti TK1 peptide antibodies, that was used to measure TK1 protein levels in dog sera. TK1 protein levels were significantly higher in sera from dogs with haematological tumours as well as solid tumours in comparison with healthy dogs. Overall, the results demonstrate that TK1 protein assays provide valuable diagnostic information in a variety of canine malignancies.

Authors/Creators:Jagarlamudi, Kiran Kumar
Title:Diagnostic implications of the molecular forms and levels of serum thymidine kinase 1 in different canine malignancies
Series/Journal:Acta Universitatis agriculturae Sueciae (1652-6880)
Year of publishing :16 March 2015
Depositing date:16 March 2015
Volume:2015:32
Number of Pages:87
Papers/manuscripts:
NumberReferences
IKiran Kumar J., Sharif H., Westberg S., von Euler H., Eriksson S., 2013. High levels of inactive thymidine kinase 1 polypeptide detected in sera from dogs with solid tumours by immunoaffinity methods: Implications for in vitro diagnostics. Vet J. 197:854-860
IIHanan S., Jagarlamudi KK., Liya W., Ellen H., Eriksson S., 2012. Quaternary structures of recombinant, cellular and serum forms of thymidine kinase 1 from dogs and humans. BMC Biochem, 13:12.
IIIJagarlamudi KK., Westberg S., Rönnberg H., Eriksson S., 2014. Properties of cellular and serum forms of thymidine kinase 1 (TK1) in dogs with acute lymphocytic leukemia (ALL) and canine mammary tumours (CMTs): Implications for TK1 as a proliferation biomarker. BMC vet Res. 10: 228.
IVKiran Kumar J., Laura M., Westberg S., Rönnberg H., Eriksson S., 2015. Development of a new ELISA for determining serum thymidine kinase 1 protein levels in canine malignancies and its clinical significance. (Manuscript).
Place of Publication:Uppsala
Publisher:Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences
ISBN for printed version:978-91-576-8262-8
ISBN for electronic version:978-91-576-8263-5
ISSN:1652-6880
Language:English
Publication Type:Doctoral thesis
Full Text Status:Public
Agris subject categories.:L Animal production > L73 Animal diseases
Subjects:(A) Swedish standard research categories 2011 > 4 Agricultural Sciences > 403 Veterinary Science > Medical Bioscience
Agrovoc terms:dogs, mammary glands, neoplasms, animal diseases, enzymes, thymidine, kinases, genetic markers, antibodies, elisa
Keywords:Thymidine kinase 1, Mammary tumors, dog TK1-ELISA
URN:NBN:urn:nbn:se:slu:epsilon-e-2494
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-e-2494
ID Code:11995
Faculty:VH - Faculty of Veterinary Medicine and Animal Science
Department:(VH) > Dept. of Anatomy, Physiology and Biochemistry
Deposited By: Dr Kiran Kumar Jagarlamudi
Deposited On:13 Mar 2015 14:37
Metadata Last Modified:31 Mar 2015 13:19

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