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Influence of BLV Infection on Telomere Length and Telomerase Activity in Cattle
114
ESVP/ECVP Proceedings 2013
150:1, 2014
DIFFUSE IRIS MELANOMA WITH ATYPICAL METASTATIC BEHAVIOUR IN A CAT A. Calado *, T. Silva *, C. Santos *, A.C. Figueira *, J. Prada y and I. Canavarro*,z *Escola Universitaria Vasco da Gama, Coimbra, yUniversidade de Tras-osMontes e Alto Douro, Vila Real and zClınica Veterinaria Vetpovoa, Coimbra, Portugal Introduction: A 13-year-old, female DSH cat was treated surgically (enucleation) for a tumour of the right eye diagnosed as diffuse iris melanoma (DIM). Fourteen months later, abdominal ultrasound revealed an unidentified mass. At laparotomy, numerous pigmented lesions were identified and the cat was humanely destroyed. Materials and Methods: A full necropsy examination was performed. Results: Darkly pigmented lesions were present in multiple organs and tissues including lymph nodes, parotid salivary gland, lungs, pleural fat, rib, diaphragm, mesentery, spleen, liver and pancreas. Microscopically, these were composed of round neoplastic melanocytes, ranging from amelanotic to highly pigmented cells. The cells were pleomorphic, with a vesicular nucleus with large prominent nucleoli. Supporting stromal tissue was sparse. Neoplastic emboli were present in peripancreatic lymphatics and in lymph node sinuses. Conclusions: A diagnosis of DIM with metastasis was made. DIM is a common primary intraocular tumour in old cats. The prognosis is poor despite enucleation, due to the high metastatic potential. Metastasis by haematogenous spread often involves the liver, kidneys and lungs. In this case there was wider dissemination throughout multiple organs. Cavitary and lymphatic spread was evident in addition to haematogenous spread.
IMMUNE CELLS INFILTRATE FELINE ENDOMETRIAL ADENOCARCINOMAS IN THE PRESENCE OR ABSENCE OF PYOMETRA M. Tavares Pereira *, A.L. Saraiva *,y, A. Gama *, R. PayanCarreira * and M.A. Pires* *CECAV-ECAV-UTAD, Vila Real and yEUVG, Coimbra, Portugal Introduction: Immune cells play important roles in tumourigenesis through the establishment of a dynamic relationship. In cats endometrial adenocarcinomas (EAs) are commonly associated with pyometra. The aim of this study was to assess the immune cell infiltrate in EA with or without concurrent pyometra. Materials and Methods: Ten samples of feline EA were evaluated (five presenting with pyometra and five without inflammation) as well as 10 control samples of endometrium. Indirect immunohistochemistry was performed using antibodies specific for macrophages (MAC387, Serotec, 1 in 100 dilution) and T (CD3, Dako, 1 in 50 dilution) and B (CD79, Cell Marque, 1 in 50 dilution) lymphocytes. Cells were counted in two different areas of the tumours and also in the surrounding tissues. Results: Immune cell infiltration was significantly increased in tumours (P !0.001). T cells were increased five-fold in tumours, whether or not there was concurrent pyometra. In contrast, B cells were slightly increased in tumours without pyometra, but showed a marked increase with inflammation. Macrophages showed only a two-fold increase in tumours compared with controls, yet an eightfold increase in macrophages was found in cases of co-existing pyometra. Conclusions: The inflammatory infiltrate differed in tumours with and without pyometra. In the absence of pyometra, T cells were the predominant population, but if pyometra was present B cells dominated.
INFLUENCE OF BLV INFECTION ON TELOMERE LENGTH AND TELOMERASE ACTIVITY IN CATTLE M. Szczotka, J. Kuzmak and E. Iwan National Veterinary Research Institute, Department of Biochemistry, Pulawy, Poland Introduction: Bovine leukaemia virus (BLV) is a causative agent of bovine leukaemia. The aim of this study was to determine telomerase activity and telomere length in bovine lymphoid cells. Telomerase, a telomere-synthesizing reverse transcriptase, compensates for the loss of telomeres associated with cell division. Telomeres are important structures for the correct function and stability of chromosomes. Telomerase activity is expressed in most human tumour tissues, but not in normal tissues, except for those of the germline (i.e. ovaries and testes). Materials and Methods: Telomerase activity was determined by use of a PCR-ELISA method in sera, lymphoid cells and dendritic cells generated from lymphoid tissue. The telomere length and fluorescence intensity was determined in interphase nuclei and metaphase spreads with the use of fluorescence in-situ hybridization (FISH) and flow cytometry. Human tumour cell lines were used as positive controls. Results: Very high relative telomerase activities were found in dendritic cells and sera from cattle with lymphocytosis and in the FLKBLV cell line. The relative telomere length was much shorter in leukaemic animals. Conclusions: High telomerase activity and shortened telomeres almost always correlate with disease severity in lymphoproliferative disorders. Measurement of telomere length and telomerase activity might be practical to monitor disease and as a prognostic marker and therapeutic target.
DETECTION AND QUANTIFICATION OF BPV-2 DNA IN BOVINE URINARY BLADDER LESIONS FROM THE AZORES ARCHIPELAGO, PORTUGAL J. Cota *, A. Duarte *, T. Carvalho y, C. Pinto z, L. Tavares * and M.C. Peleteiro* *CIISA, Faculdade de Medicina Veterinaria, Universidade Tecnica de Lisboa, y IMM, Faculdade de Medicina, Universidade de Lisboa and zServic¸os de Desenvolvimento Agrario, Ponta Delgada, Portugal Introduction: Bovine papillomavirus type 2 (BPV-2) DNA has been identified in the urinary bladder (UB) of animals suffering from bovine enzootic haematuria (BEH). Few reports have given information on the viral load. The aim of this study was to quantify the number of BPV-2 virions in animals slaughtered in the Azores Archipelago, a BEH endemic region. Materials and Methods: Fifty-one animals were tested using a realtime PCR system for detection and quantification of BPV-2. Thirtyeight animals had gross lesions in the UB and 13 had no lesions. The iliac lymph nodes of 35 animals were also tested. Results: BPV-2 DNA was detected in 27 of the 38 UBs with lesions (71%), in seven of the 13 bladders with no lesions (54%) and in seven of the 35 lymph nodes (20%). The highest number of BPV-2 DNA copies was 4.2/cell and the lowest was 0.0002/cell, with an average of 0.08 copies/cell. Conclusions: This is the first report on the quantification of BPV-2 in UB lesions from animals of the Azores Archipelago.
ESVP/ECVP Proceedings 2013
150:1, 2014
DIFFUSE IRIS MELANOMA WITH ATYPICAL METASTATIC BEHAVIOUR IN A CAT A. Calado *, T. Silva *, C. Santos *, A.C. Figueira *, J. Prada y and I. Canavarro*,z *Escola Universitaria Vasco da Gama, Coimbra, yUniversidade de Tras-osMontes e Alto Douro, Vila Real and zClınica Veterinaria Vetpovoa, Coimbra, Portugal Introduction: A 13-year-old, female DSH cat was treated surgically (enucleation) for a tumour of the right eye diagnosed as diffuse iris melanoma (DIM). Fourteen months later, abdominal ultrasound revealed an unidentified mass. At laparotomy, numerous pigmented lesions were identified and the cat was humanely destroyed. Materials and Methods: A full necropsy examination was performed. Results: Darkly pigmented lesions were present in multiple organs and tissues including lymph nodes, parotid salivary gland, lungs, pleural fat, rib, diaphragm, mesentery, spleen, liver and pancreas. Microscopically, these were composed of round neoplastic melanocytes, ranging from amelanotic to highly pigmented cells. The cells were pleomorphic, with a vesicular nucleus with large prominent nucleoli. Supporting stromal tissue was sparse. Neoplastic emboli were present in peripancreatic lymphatics and in lymph node sinuses. Conclusions: A diagnosis of DIM with metastasis was made. DIM is a common primary intraocular tumour in old cats. The prognosis is poor despite enucleation, due to the high metastatic potential. Metastasis by haematogenous spread often involves the liver, kidneys and lungs. In this case there was wider dissemination throughout multiple organs. Cavitary and lymphatic spread was evident in addition to haematogenous spread.
IMMUNE CELLS INFILTRATE FELINE ENDOMETRIAL ADENOCARCINOMAS IN THE PRESENCE OR ABSENCE OF PYOMETRA M. Tavares Pereira *, A.L. Saraiva *,y, A. Gama *, R. PayanCarreira * and M.A. Pires* *CECAV-ECAV-UTAD, Vila Real and yEUVG, Coimbra, Portugal Introduction: Immune cells play important roles in tumourigenesis through the establishment of a dynamic relationship. In cats endometrial adenocarcinomas (EAs) are commonly associated with pyometra. The aim of this study was to assess the immune cell infiltrate in EA with or without concurrent pyometra. Materials and Methods: Ten samples of feline EA were evaluated (five presenting with pyometra and five without inflammation) as well as 10 control samples of endometrium. Indirect immunohistochemistry was performed using antibodies specific for macrophages (MAC387, Serotec, 1 in 100 dilution) and T (CD3, Dako, 1 in 50 dilution) and B (CD79, Cell Marque, 1 in 50 dilution) lymphocytes. Cells were counted in two different areas of the tumours and also in the surrounding tissues. Results: Immune cell infiltration was significantly increased in tumours (P !0.001). T cells were increased five-fold in tumours, whether or not there was concurrent pyometra. In contrast, B cells were slightly increased in tumours without pyometra, but showed a marked increase with inflammation. Macrophages showed only a two-fold increase in tumours compared with controls, yet an eightfold increase in macrophages was found in cases of co-existing pyometra. Conclusions: The inflammatory infiltrate differed in tumours with and without pyometra. In the absence of pyometra, T cells were the predominant population, but if pyometra was present B cells dominated.
INFLUENCE OF BLV INFECTION ON TELOMERE LENGTH AND TELOMERASE ACTIVITY IN CATTLE M. Szczotka, J. Kuzmak and E. Iwan National Veterinary Research Institute, Department of Biochemistry, Pulawy, Poland Introduction: Bovine leukaemia virus (BLV) is a causative agent of bovine leukaemia. The aim of this study was to determine telomerase activity and telomere length in bovine lymphoid cells. Telomerase, a telomere-synthesizing reverse transcriptase, compensates for the loss of telomeres associated with cell division. Telomeres are important structures for the correct function and stability of chromosomes. Telomerase activity is expressed in most human tumour tissues, but not in normal tissues, except for those of the germline (i.e. ovaries and testes). Materials and Methods: Telomerase activity was determined by use of a PCR-ELISA method in sera, lymphoid cells and dendritic cells generated from lymphoid tissue. The telomere length and fluorescence intensity was determined in interphase nuclei and metaphase spreads with the use of fluorescence in-situ hybridization (FISH) and flow cytometry. Human tumour cell lines were used as positive controls. Results: Very high relative telomerase activities were found in dendritic cells and sera from cattle with lymphocytosis and in the FLKBLV cell line. The relative telomere length was much shorter in leukaemic animals. Conclusions: High telomerase activity and shortened telomeres almost always correlate with disease severity in lymphoproliferative disorders. Measurement of telomere length and telomerase activity might be practical to monitor disease and as a prognostic marker and therapeutic target.
DETECTION AND QUANTIFICATION OF BPV-2 DNA IN BOVINE URINARY BLADDER LESIONS FROM THE AZORES ARCHIPELAGO, PORTUGAL J. Cota *, A. Duarte *, T. Carvalho y, C. Pinto z, L. Tavares * and M.C. Peleteiro* *CIISA, Faculdade de Medicina Veterinaria, Universidade Tecnica de Lisboa, y IMM, Faculdade de Medicina, Universidade de Lisboa and zServic¸os de Desenvolvimento Agrario, Ponta Delgada, Portugal Introduction: Bovine papillomavirus type 2 (BPV-2) DNA has been identified in the urinary bladder (UB) of animals suffering from bovine enzootic haematuria (BEH). Few reports have given information on the viral load. The aim of this study was to quantify the number of BPV-2 virions in animals slaughtered in the Azores Archipelago, a BEH endemic region. Materials and Methods: Fifty-one animals were tested using a realtime PCR system for detection and quantification of BPV-2. Thirtyeight animals had gross lesions in the UB and 13 had no lesions. The iliac lymph nodes of 35 animals were also tested. Results: BPV-2 DNA was detected in 27 of the 38 UBs with lesions (71%), in seven of the 13 bladders with no lesions (54%) and in seven of the 35 lymph nodes (20%). The highest number of BPV-2 DNA copies was 4.2/cell and the lowest was 0.0002/cell, with an average of 0.08 copies/cell. Conclusions: This is the first report on the quantification of BPV-2 in UB lesions from animals of the Azores Archipelago.