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Preneoplastic and Neoplastic Mammary and Non-mammary Lesions Associated with Inflammation in Chemically Induced Carcinogenesis in Wistar Rats
146:1, 2012
ESVP/ECVP Proceedings 2011
65
PRENEOPLASTIC AND NEOPLASTIC MAMMARY AND NON-MAMMARY LESIONS ASSOCIATED WITH INFLAMMATION IN CHEMICALLY INDUCED CARCINOGENESIS IN WISTAR RATS A. F. Gal, S. Andrei, C. Bouari, M. Taulescu, P. Bolfa and C. C atoi Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, Romania Introduction: Recent data have expanded the concept that inflammation is an intrinsic participant in the neoplastic process, fostering proliferation, survival and migration. The aim of this study was to follow up the incidence of chronic inflammation in chemically-induced mammary carcinogenesis. Materials and Methods: Three groups of 37-day-old female Wistar rats were studied: group I inoculated with N-methyl-N-nitroso-urea (MNU), group II given MNU+astaxanthin (ASTA) in the diet and group III given ASTA. ASTA was administered orally (50 mg astaxanthin/rat/day for 7 months). The experiment was ended 14 months after MNU intake. Samples harvested for histopathological examination were processed to paraffin wax. Results: Mammary tumour induction by MNU was reduced (33.3e37.5%). Several other tumour types were diagnosed in other organs (cholangiocarcinoma, nephroblastoma and lung carcinoma). Both groups inoculated with MNU encountered precancerous hyperplastic mammary lesions (simple adenosis, typical lobular epitheliosis), but there was no inflammation in the mammary parenchyma. Chronic inflammation has been associated with hyperplasia and/or cancer in the lungs, liver and kidneys. Conclusions: A strong association between chronic inflammation and MNU-induced carcinogenesis exists. The chronic inflammatory state may lead to environments that foster genomic lesions and tumour initiation.
TELOMERASE ACTIVITY IN CATTLE INFECTED WITH BOVINE LEUKAEMIA VIRUS (BLV) M. Szczotka and J. Kuzmak Department of Biochemistry, National Veterinary Research Institute, Pulawy, Poland Introduction: Telomerase is a telomere synthesizing reverse transcriptase. This enzyme compensates for the loss of telomere associated with cell division. Telomerase adds new telomeric sequences to the end of chromosomal DNA in order to overcome the end-replicating problem. In man and other vertebrates, the telomeric sequence contains TTAGGG repeats. Telomerase activity is present in embryonal and germ cells, but is undetectable in most somatic cells, although many tumour cells have a high level of telomerase activity. Telomerase reactivation in tumour cells has been observed in some mammals. Therefore, telomerase activity has been proposed as a tumour marker in these animals. The bovine leukaemia virus (BLV) is an oncogenic B-lymphotropic retrovirus that causes enzootic bovine leukaemia, the most common tumour in cattle. Materials and Methods: Investigations were performed on a group of 80 cattle infected with BLV and specific antibodies and proviral DNA were detected in their sera by ELISA and PCR, respectively. Telomerase activity was measured in sera, plasma and lysates of blood lymphocytes, spleen, lymph nodes, bone marrow and supernatants of these cells, cultured in vitro. The same investigations were performed with materials taken from 21 healthy control cows. Telomerase activity was determined with the use of a commercial ELISA kit (Cusabio). Results: The concentrations of telomerase in the sera of BLV-infected cows were 0.119e0.354 ng/ml. In the plasma, the telomerase concentrations were 0.105e0.279 ng/ml. In supernatants from lymphoid cells cultured in vitro these concentrations were 0.177e0.482 ng/ml. In samples from control animals telomerase activity was undetectable. Conclusions: Similar to many human tumours, telomerase activity was detected in cows infected with bovine leukaemia virus and this activity may be a useful marker for tumour development or act as a potential therapeutic target.
EXPERIMENTAL STUDIES OF PATHOGENECITY OF CHICKEN INFECTIOUS ANAEMIA VIRUS (THREE ISOLATES) IN IRAN A. Ezzi, A. Shoushtari and H. Mardjanmehr Razi Vaccine and Serum Research Institute, Karaj, Iran Introduction: Chicken anaemia virus (CAV) is a small non-enveloped icosahedral virus with a negative sense, single-stranded circular DNA genome. It has been classified as the only member of the genus Gyrovirus of the family Giroviridae. Materials and Methods: The aim of this experiment was to evaluate pathogenicity of three CAV isolates, CV1, CV2 and CV3. Thirty 1-day-old SPF chickens were grouped and received an intramuscular inoculation of one isolate per group. Two other groups (control groups) were inoculated with a live vaccine virus and normal saline, respectively. The packed cell volumes (PCVs) were determined on blood samples from each bird. Antibodies were measured by competitive ELISA. Samples of liver, bursa of Fabricius, spleen, thymus and skeletal muscle were fixed in 10% neutral buffered formalin, processed and embedded in paraffin wax. The blocks were sectioned (5 mm) and stained with haematoxylin and eosin. The lesions of the bursa and thymus were evaluated for lymphocyte depletion and scored as: 1, normal; 2, mild; 3, moderate and 4, severe. Results: Birds in the first three groups showed ruffled feathers, depression and body weight reduction. After 18 days they were weighed, bled and killed. Three birds were found dead during the experiment (one in each test group). PCVs of the three tested groups were below normal. Grossly the thymus and bursa were severely atrophic. Bone marrow was yellow and pale. Severe atrophy and depletion of the thymus, bursa of Fabricius and bone marrow was observed and this was significantly different to control groups (P ! 0.05). Conclusions: While CAV infection is understood to be most pathogenic in young growing birds, until now the infection has only been traced in slaughter-age chickens in Iran. The present work showed the pathogenicity of CAV in 1-day-old chickens and displayed the detrimental impacts of CAV on the immune system of chickens with apparent concentration in thymus.
IMMUNOHISTOCHEMICAL CHARACTERISATION OF IMMUNE CELL SUBSETS IN LYMPH NODES FROM WATER BUFFALOES G. Cant on, F. Chianini, J. L. Konrad and C. M. Campero Moredun Research Institute, Scotland and INTA, Argentina Introduction: Water buffaloes (Bubalus bubalis) play a crucial role in Asian agriculture and their importance is increasing in western nations. They are susceptible to similar aetiological agents of disease as cattle, but the outcome may be different. This may be due to differences in the responses of the immune systems in the two different species. The aim of this work was to characterize immune system cell subsets in fixed lymph nodes from buffaloes. Materials and Methods: Immunohistochemistry was performed on zinc salts fixed paraffin-wax embedded lymph nodes from healthy water buffalo. Monoclonal antibodies (mAbs) were selected from those used in other species or reported previously for water buffalo tissues using other techniques. Results: Specific labelling was observed using mAbs previously unreported as cross-reacting with buffalo tissues: EBM11 (macrophages), CC58 (CD8 T cells), IL-A29 (gdTCR), NKp46 (NK cells) and HM57 (B cells)] or using clones previously described for use in flow cytometry: MMIA (CD3 T cells), IL-A11 and CC30 (CD4 T-cells). Conclusions: The results from this study provide a new panel of mAbs to investigate the buffalo inflammatory response to diseases in fixed tissues. Other mAbs previously used in ruminants could also be examined to provide further tools for use with water buffalo tissues.
ESVP/ECVP Proceedings 2011
65
PRENEOPLASTIC AND NEOPLASTIC MAMMARY AND NON-MAMMARY LESIONS ASSOCIATED WITH INFLAMMATION IN CHEMICALLY INDUCED CARCINOGENESIS IN WISTAR RATS A. F. Gal, S. Andrei, C. Bouari, M. Taulescu, P. Bolfa and C. C atoi Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, Romania Introduction: Recent data have expanded the concept that inflammation is an intrinsic participant in the neoplastic process, fostering proliferation, survival and migration. The aim of this study was to follow up the incidence of chronic inflammation in chemically-induced mammary carcinogenesis. Materials and Methods: Three groups of 37-day-old female Wistar rats were studied: group I inoculated with N-methyl-N-nitroso-urea (MNU), group II given MNU+astaxanthin (ASTA) in the diet and group III given ASTA. ASTA was administered orally (50 mg astaxanthin/rat/day for 7 months). The experiment was ended 14 months after MNU intake. Samples harvested for histopathological examination were processed to paraffin wax. Results: Mammary tumour induction by MNU was reduced (33.3e37.5%). Several other tumour types were diagnosed in other organs (cholangiocarcinoma, nephroblastoma and lung carcinoma). Both groups inoculated with MNU encountered precancerous hyperplastic mammary lesions (simple adenosis, typical lobular epitheliosis), but there was no inflammation in the mammary parenchyma. Chronic inflammation has been associated with hyperplasia and/or cancer in the lungs, liver and kidneys. Conclusions: A strong association between chronic inflammation and MNU-induced carcinogenesis exists. The chronic inflammatory state may lead to environments that foster genomic lesions and tumour initiation.
TELOMERASE ACTIVITY IN CATTLE INFECTED WITH BOVINE LEUKAEMIA VIRUS (BLV) M. Szczotka and J. Kuzmak Department of Biochemistry, National Veterinary Research Institute, Pulawy, Poland Introduction: Telomerase is a telomere synthesizing reverse transcriptase. This enzyme compensates for the loss of telomere associated with cell division. Telomerase adds new telomeric sequences to the end of chromosomal DNA in order to overcome the end-replicating problem. In man and other vertebrates, the telomeric sequence contains TTAGGG repeats. Telomerase activity is present in embryonal and germ cells, but is undetectable in most somatic cells, although many tumour cells have a high level of telomerase activity. Telomerase reactivation in tumour cells has been observed in some mammals. Therefore, telomerase activity has been proposed as a tumour marker in these animals. The bovine leukaemia virus (BLV) is an oncogenic B-lymphotropic retrovirus that causes enzootic bovine leukaemia, the most common tumour in cattle. Materials and Methods: Investigations were performed on a group of 80 cattle infected with BLV and specific antibodies and proviral DNA were detected in their sera by ELISA and PCR, respectively. Telomerase activity was measured in sera, plasma and lysates of blood lymphocytes, spleen, lymph nodes, bone marrow and supernatants of these cells, cultured in vitro. The same investigations were performed with materials taken from 21 healthy control cows. Telomerase activity was determined with the use of a commercial ELISA kit (Cusabio). Results: The concentrations of telomerase in the sera of BLV-infected cows were 0.119e0.354 ng/ml. In the plasma, the telomerase concentrations were 0.105e0.279 ng/ml. In supernatants from lymphoid cells cultured in vitro these concentrations were 0.177e0.482 ng/ml. In samples from control animals telomerase activity was undetectable. Conclusions: Similar to many human tumours, telomerase activity was detected in cows infected with bovine leukaemia virus and this activity may be a useful marker for tumour development or act as a potential therapeutic target.
EXPERIMENTAL STUDIES OF PATHOGENECITY OF CHICKEN INFECTIOUS ANAEMIA VIRUS (THREE ISOLATES) IN IRAN A. Ezzi, A. Shoushtari and H. Mardjanmehr Razi Vaccine and Serum Research Institute, Karaj, Iran Introduction: Chicken anaemia virus (CAV) is a small non-enveloped icosahedral virus with a negative sense, single-stranded circular DNA genome. It has been classified as the only member of the genus Gyrovirus of the family Giroviridae. Materials and Methods: The aim of this experiment was to evaluate pathogenicity of three CAV isolates, CV1, CV2 and CV3. Thirty 1-day-old SPF chickens were grouped and received an intramuscular inoculation of one isolate per group. Two other groups (control groups) were inoculated with a live vaccine virus and normal saline, respectively. The packed cell volumes (PCVs) were determined on blood samples from each bird. Antibodies were measured by competitive ELISA. Samples of liver, bursa of Fabricius, spleen, thymus and skeletal muscle were fixed in 10% neutral buffered formalin, processed and embedded in paraffin wax. The blocks were sectioned (5 mm) and stained with haematoxylin and eosin. The lesions of the bursa and thymus were evaluated for lymphocyte depletion and scored as: 1, normal; 2, mild; 3, moderate and 4, severe. Results: Birds in the first three groups showed ruffled feathers, depression and body weight reduction. After 18 days they were weighed, bled and killed. Three birds were found dead during the experiment (one in each test group). PCVs of the three tested groups were below normal. Grossly the thymus and bursa were severely atrophic. Bone marrow was yellow and pale. Severe atrophy and depletion of the thymus, bursa of Fabricius and bone marrow was observed and this was significantly different to control groups (P ! 0.05). Conclusions: While CAV infection is understood to be most pathogenic in young growing birds, until now the infection has only been traced in slaughter-age chickens in Iran. The present work showed the pathogenicity of CAV in 1-day-old chickens and displayed the detrimental impacts of CAV on the immune system of chickens with apparent concentration in thymus.
IMMUNOHISTOCHEMICAL CHARACTERISATION OF IMMUNE CELL SUBSETS IN LYMPH NODES FROM WATER BUFFALOES G. Cant on, F. Chianini, J. L. Konrad and C. M. Campero Moredun Research Institute, Scotland and INTA, Argentina Introduction: Water buffaloes (Bubalus bubalis) play a crucial role in Asian agriculture and their importance is increasing in western nations. They are susceptible to similar aetiological agents of disease as cattle, but the outcome may be different. This may be due to differences in the responses of the immune systems in the two different species. The aim of this work was to characterize immune system cell subsets in fixed lymph nodes from buffaloes. Materials and Methods: Immunohistochemistry was performed on zinc salts fixed paraffin-wax embedded lymph nodes from healthy water buffalo. Monoclonal antibodies (mAbs) were selected from those used in other species or reported previously for water buffalo tissues using other techniques. Results: Specific labelling was observed using mAbs previously unreported as cross-reacting with buffalo tissues: EBM11 (macrophages), CC58 (CD8 T cells), IL-A29 (gdTCR), NKp46 (NK cells) and HM57 (B cells)] or using clones previously described for use in flow cytometry: MMIA (CD3 T cells), IL-A11 and CC30 (CD4 T-cells). Conclusions: The results from this study provide a new panel of mAbs to investigate the buffalo inflammatory response to diseases in fixed tissues. Other mAbs previously used in ruminants could also be examined to provide further tools for use with water buffalo tissues.