Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 257
Filter
2.
Open Vet J ; 12(2): 264-272, 2022.
Article in English | MEDLINE | ID: mdl-35603075

ABSTRACT

Background: Ovine pulmonary adenocarcinoma (OPA), caused by Jaagsiekte sheep retrovirus (JSRV), is a contagious neoplastic disease in sheep characterized by chronic respiratory signs, inducing the transformation of secretory epithelial cells of the distal respiratory tract. Aims: To perform clinical, epidemiological, and molecular studies with evaluation of some predisposing factors at the herd level of OPA infection in sheep in Al-Qadisiyah Province, Iraq. Methods: The first step of the study was undertaken to evaluate the clinical cases of OPA in naturally infected sheep and correlation with observing respiratory signs. Seventy-five sheep with chronic respiratory signs were examined clinically, and by molecular and sequences analysis. The second step was the epidemiological part that was carried out on 195 randomly selected animals from 30 flocks, with the prevalence rate based on PCR; sex, age, and size of flocks were assessed, as well as macroscopic and microscopic features of the neoplastic lung. Deep nasal swabs and nasal secretion were collected from all animals. RNA extraction and RT-PCR were also carried out. Results: The results showed that 12 (16%) samples were positive for OPA, based on env gene-specific primers. Nucleotide sequences of partial 545 bp of the env gene showed (0.07-0.12) variations from global strains presented in the NCBI database. The prevalence rate of OPA was 21/195 (10.76%) with PCR. The epidemiological factors analysis showed that there was no effect of sex and herd size on the prevalence rates (p ≥ 0.01), whereas age was significantly affected and the age of 2-4 years was more susceptible (p ≥ 0.01). Gross and microscopic examinations were discussed with the confirmation of an OPA infection. Conclusion: The current study provides useful data about the clinical and epidemiological features of JSRV that is circulating in sheep of Iraq, and concludes that epidemiological studies and disease control may require multi-diagnostic assays.


Subject(s)
Jaagsiekte sheep retrovirus , Pulmonary Adenomatosis, Ovine , Sheep Diseases , Animals , Iraq/epidemiology , Jaagsiekte sheep retrovirus/genetics , Polymerase Chain Reaction/veterinary , Pulmonary Adenomatosis, Ovine/epidemiology , Pulmonary Adenomatosis, Ovine/pathology , Sheep/genetics , Sheep Diseases/epidemiology
3.
Vet Rec ; 191(3): e1578, 2022 08.
Article in English | MEDLINE | ID: mdl-35347736

ABSTRACT

BACKGROUND: Transthoracic ultrasonography (TTUS) is currently the only widely used method to diagnose preclinical or subclinical ovine pulmonary adenocarcinoma (OPA) in the live sheep. However, little is known about the test characteristics of TTUS. METHODS: One thousand and seventy-four breeding ewes in a flock with evidence of low OPA prevalence underwent TTUS by an experienced operator. Fifty-one sheep were diagnosed with OPA and underwent gross postmortem examination (PME). RESULTS: Lesions consistent with OPA were found in only 24% (12/51) of the culled ewes. Thirty-five percent (18/51) of culled ewes had gross lesions consistent with other pulmonary disease and 41% (21/51) had no detectable gross lesions on PME. Histopathology and immunohistochemistry confirmed OPA in only the 12 animals identified with OPA lesions from PME. CONCLUSION: Great caution should be exercised when deciding if TTUS is an appropriate screening test in groups of sheep where OPA prevalence may be anticipated to be low. TTUS is a subjective test and thus individual operator ability will influence the sensitivity and specificity of TTUS for OPA diagnosis while the underlying prevalence influences the eventual positive predictive value.


Subject(s)
Adenocarcinoma of Lung , Lung Neoplasms , Pulmonary Adenomatosis, Ovine , Adenocarcinoma of Lung/veterinary , Animals , Female , Jaagsiekte sheep retrovirus , Lung Neoplasms/diagnostic imaging , Lung Neoplasms/veterinary , Pulmonary Adenomatosis, Ovine/diagnostic imaging , Pulmonary Adenomatosis, Ovine/epidemiology , Sheep , Sheep Diseases , Ultrasonography/veterinary , United Kingdom/epidemiology
4.
Arch Virol ; 166(3): 831-840, 2021 Mar.
Article in English | MEDLINE | ID: mdl-33486631

ABSTRACT

Ovine pulmonary adenomatosis (OPA) is caused by jaagsiekte sheep retrovirus (JSRV) and is a chronic, progressive, and infectious neoplastic lung disease in sheep, which causes significant economic losses to the sheep industry. Neither a vaccine nor serological diagnostic methods to detect OPA are available. We performed a JSRV infection survey in sheep using blood samples (n = 1,372) collected in the three northeastern provinces of China (i.e., Inner Mongolia, Heilongjiang, and Jilin) to determine JSRV infection status in sheep herds using a real-time PCR assay targeting the gag gene of JSRV. The ovine endogenous retrovirus sequence was successfully amplified in all sheep samples tested (296 from the Inner Mongolia Autonomous Region, 255 from Jilin province, and 821 from Heilongjiang province). Subsequently, we attempted to distinguish exogenous JSRV (exJSRV) and endogenous JSRV (enJSRV) infections in these JSRV-positive samples using a combination assay that identifies a ScaI restriction site in an amplified 229-bp fragment of the gag gene of JSRV and a "LHMKYXXM" motif in the cytoplasmic tail region of the JSRV envelope protein. The ScaI restriction site is present in all known oncogenic JSRVs but absent in ovine endogenous retroviruses, while the "LHMKYXXM" motif is in all known exJSRVs but not in enJSRVs. Interestingly, one JSRV strain (HH13) from Heilongjiang province contained the "LHMKYXXM" motif but not the ScaI enzyme site. Phylogenetic analysis showed that strain HH13 was closely related to strain enJSRV-21 reported in the USA, indicating that HH13 could be an exogenous virus. Our results provide valuable information for further research on the genetic evolution and pathogenesis of JSRV.


Subject(s)
Endogenous Retroviruses/genetics , Gene Products, env/genetics , Jaagsiekte sheep retrovirus/genetics , Pulmonary Adenomatosis, Ovine/epidemiology , Pulmonary Adenomatosis, Ovine/pathology , Amino Acid Motifs/genetics , Animals , Base Sequence , China/epidemiology , DNA, Viral/analysis , Deoxyribonucleases, Type II Site-Specific/metabolism , Evolution, Molecular , Genome, Viral/genetics , Jaagsiekte sheep retrovirus/classification , Phylogeny , Real-Time Polymerase Chain Reaction , Restriction Mapping , Sequence Homology, Nucleic Acid , Sheep
5.
Vet Pathol ; 58(2): 361-368, 2021 03.
Article in English | MEDLINE | ID: mdl-33357120

ABSTRACT

Contagious respiratory tumors of sheep and goats are epithelial neoplasms of the lung and nasal cavities. They are associated with oncogenic betaretroviruses known as jaagsiekte sheep retrovirus and enzootic nasal tumor retrovirus of sheep and goats. We investigated the presence of the envelope protein (ENV) of these retroviruses in retropharyngeal and mediastinal lymph nodes using a specific monoclonal antibody by immunohistochemistry methods, single-labeled or combined with ovine B or T lymphocytes or macrophage cell markers. Samples of lymph nodes, fixed in formalin and zinc fixative, were obtained from paraffin-embedded material. Four groups of samples were used: 24 natural cases of ovine pulmonary adenocarcinoma (OPA), 13 of enzootic nasal adenocarcinoma of sheep (ENAS), 19 of enzootic nasal adenocarcinoma of goats (ENAG), and 14 control samples. ENV was detected by single labeling in cortical lymphoid follicles. Six of 24 OPA samples were positive and only in those from sheep with extensive neoplasia. Immunolabeling was detected in 5/13 ENAS and 10/19 ENAG samples. Positive labeling was found either in the intercellular spaces, membranes, or cytoplasm of cells in follicles. Control samples were not correspondingly labeled. Double immunohistochemistry demonstrated co-labeling of ENV and CD21 (B cells and follicular dendritic cells) in all samples, CD14 (macrophage) in OPA samples, and Pax-5 (B cells) in ENAG samples, but not with CD8 or CD4 (T lymphocytes). These results demonstrate the presence of betaretrovirus ENV proteins in nontumor cells in regional lymph nodes in sheep and goats with contagious respiratory tumors.


Subject(s)
Betaretrovirus , Goat Diseases , Jaagsiekte sheep retrovirus , Pulmonary Adenomatosis, Ovine , Sheep Diseases , Animals , Goats , Lymph Nodes , Ruminants , Sheep
6.
Virus Genes ; 57(1): 50-59, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33151445

ABSTRACT

Enzootic nasal tumor virus type 1 (ENTV-1) (ovine nasal tumor virus) and ENTV-2 (caprine nasal tumor virus) are known to be causative agents of enzootic nasal adenocarcinoma (ENA) in sheep and goats, respectively. Although the nucleotide and amino acid sequences of ENTV-1 and ENTV-2 are quite similar, they are recognized as phylogenetically distinct viruses. The envelope protein of ENTV-1 functions as an oncoprotein in the in vitro transformation of epithelial cells and fibroblasts. Thus, it is the primary determinant of in vivo tumorigenesis in ENA. As per our knowledge, no previous studies have reported in detail the role of ENTV-2 in ENA tumorigenesis. Here, in order to investigate the molecular mechanism of caprine ENA oncogenesis by ENTV-2, we have attempted to identify the transforming potential of ENTV-2 envelope, and investigated the activation of cell signaling pathways in oncogenic transformation. Our findings confirmed that ENTV-2 envelope was capable of inducing oncogenic transformation of rat cell lines in vitro. Further, we found that MAPK, Akt, and p38 were constitutively activated in ENTV-2 envelope-transformed clone cells. In addition, inhibitor experiments revealed that MEK-MAPK and PI3K-Akt signaling pathways are involved in the ENTV-2 envelope-induced cell transformation. These data indicate that ENTV-2 envelope could induce oncogenic transformation by signaling pathways that are also utilized by ENTV-1 envelope.


Subject(s)
Cell Transformation, Viral , Gene Products, env/metabolism , Jaagsiekte sheep retrovirus/pathogenicity , Pulmonary Adenomatosis, Ovine/virology , Tumor Virus Infections/virology , Amino Acid Sequence , Animals , Cell Line , Epithelial Cells , Fibroblasts , HEK293 Cells , Humans , Rats , Sheep , Signal Transduction
7.
BMC Vet Res ; 16(1): 296, 2020 Aug 17.
Article in English | MEDLINE | ID: mdl-32807166

ABSTRACT

BACKGROUND: Ovine pulmonary adenocarcinoma (OPA) is a neoplastic disease caused by exogenous Jaagsiekte Sheep Retrovirus (exJSRV). The prevalence of JSRV-related OPA in Eastern European countries, including Romania is unknown. We aimed to investigate: the prevalence and morphological features of OPA (classical and atypical forms) in the Transylvania region (Romania), the immunophenotype of the pulmonary tumors and their relationships with exJSRV infection. A total of 2693 adult ewes slaughtered between 2017 and 2019 in two private slaughterhouses from Transylvania region (Romania) was evaluated. Lung tumors were subsequently assessed by cytology, histology, immunocytochemistry, immunohistochemistry, electron microscopy and DNA testing. RESULTS: Out of 2693 examined sheep, 34 had OPA (1.26% prevalence). The diaphragmatic lobes were the most affected. Grossly, the classical OPA was identified in 88.24% of investigated cases and the atypical OPA in 11.76% that included solitary myxomatous nodules. Histopathology results confirmed the presence of OPA in all suspected cases, which were classified into acinar and papillary types. Myxoid growths (MGs) were diagnosed in 6 classical OPA cases and in 2 cases of atypical form. Lung adenocarcinoma was positive for MCK and TTF-1, and MGs showed immunoreaction for Vimentin, Desmin and SMA; Ki67 expression of classical OPA was higher than atypical OPA and MGs. JSRV-MA was identified by IHC (94.11%) in both epithelial and mesenchymal cells of OPA. Immunocytochemistry and electron microscopy also confirmed the JSRV within the neoplastic cells. ExJSRV was identified by PCR in 97.05% of analyzed samples. Phylogenetic analysis revealed the presence of the exJSRV type 2 (MT809678.1) in Romanian sheep affected by lung cancer and showed a high similarity with the UK strain (AF105220.1). CONCLUSIONS: In this study, we confirmed for the first time in Romania the presence of exJSRV in naturally occurring OPA in sheep. Additionally, we described the first report of atypical OPA in Romania, and to the best of our knowledge, in Eastern Europe. Finally, we showed that MGs have a myofibroblastic origin.


Subject(s)
Adenocarcinoma of Lung/veterinary , Jaagsiekte sheep retrovirus/isolation & purification , Lung Neoplasms/veterinary , Pulmonary Adenomatosis, Ovine/epidemiology , Adenocarcinoma of Lung/pathology , Adenocarcinoma of Lung/virology , Animals , Female , Immunohistochemistry/veterinary , Immunophenotyping/veterinary , Jaagsiekte sheep retrovirus/classification , Jaagsiekte sheep retrovirus/ultrastructure , Lung Neoplasms/pathology , Lung Neoplasms/virology , Microscopy, Electron/veterinary , Phylogeny , Prevalence , Romania/epidemiology , Sheep , Sheep, Domestic
8.
J Virol Methods ; 284: 113923, 2020 10.
Article in English | MEDLINE | ID: mdl-32615131

ABSTRACT

Ovine pulmonary adenocarcinoma (OPA) is a globally occurring tumor of lung epithelium which seriously affects the development of sheep farming. In our research, lung tissues of 3 naturally infected OPA individuals and 3 healthy individuals (2-4 years old) were collected. RNA was extracted for transcriptome analysis and reference gene selection. According to transcriptome analysis, 7 candidate reference genes (eukaryotic translation initiation factor 1, EIF1; glyceraldehyde-3-phosphate dehydrogenase, GAPDH; beta-actin, ACTB; GABA Type A receptor-associated protein, GABARAP; activating transcription factor 4, ATF4; ribosomal protein S15, RPS15; and Y-Box binding protein 1, YBX1) showed fragments per kilobase of transcript per million fragments mapped (FPKM) values > 200.0 and standard errors of the means (SEM) < 20.0. Expression of the above candidate reference genes was evaluated by Real-time quantitative polymerase chain reaction (RT-qPCR) combined with the analysis using GeNorm, NormFinder, and BestKeeper software. Comprehensive analysis of the results showed that ACTB was the most stable one, followed by EIF1 and GABARAP. Then, expression stability of the above three genes were validated, suggesting as suitable reference genes in sheep lung tissue, in additional 30 OPA-affected lung tissues and 10 healthy ovine lung tissues. Finally, our findings will be helpful for the subsequent study on the tumorigenic mechanism of OPA.


Subject(s)
Gene Expression Profiling/standards , Lung/metabolism , Pulmonary Adenomatosis, Ovine/metabolism , Actins/genetics , Animals , Eukaryotic Initiation Factor-1/genetics , Female , Gene Expression Profiling/methods , Jaagsiekte sheep retrovirus , Lung/pathology , Microtubule-Associated Proteins/genetics , Pulmonary Adenomatosis, Ovine/genetics , Pulmonary Adenomatosis, Ovine/pathology , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reference Standards , Reproducibility of Results , Sequence Analysis, RNA , Sheep
9.
J Vet Diagn Invest ; 32(1): 152-155, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31884891

ABSTRACT

Betaretrovirus-induced transmissible respiratory tumors in sheep arise at 2 distinct anatomic locations, either deep in the lung tissue caused by jaagsiekte sheep retrovirus (JSRV) or in the nasal cavity induced by ovine enzootic nasal tumor virus (ENTV-1). JSRV and ENTV-1 are found in many countries worldwide and have a significant economic and animal health impact. Although JSRV is endemic in sheep in the British Isles, ENTV-1 has not been reported. We report herein a nasal adenocarcinoma in a cull 8-y-old Belclare ewe from Ireland. The gross and microscopic features and immunohistochemistry results were consistent with an ENTV-1-associated tumor. However, differential PCR, using primers specific to regions of divergent sequence between the viruses, was performed on different parts of the adenocarcinoma and produced consistent results: positive for JSRV and negative for ENTV-1. An association of JSRV with nasal adenocarcinoma in sheep has not been reported previously, to our knowledge. Our case shows the necessity of using PCR in combination with immunohistochemistry to reach an accurate etiologic diagnosis, which is of importance in countries currently free of ENTV-1.


Subject(s)
Adenocarcinoma/veterinary , Jaagsiekte sheep retrovirus , Nose Neoplasms/veterinary , Pulmonary Adenomatosis, Ovine/virology , Adenocarcinoma/epidemiology , Adenocarcinoma/virology , Animals , Female , Ireland/epidemiology , Nose Neoplasms/epidemiology , Nose Neoplasms/virology , Pulmonary Adenomatosis, Ovine/epidemiology , Pulmonary Adenomatosis, Ovine/pathology , Sheep
10.
Viruses ; 11(11)2019 11 14.
Article in English | MEDLINE | ID: mdl-31739606

ABSTRACT

Jaagsiekte sheep retrovirus (JSRV) and enzootic nasal tumor virus (ENTV) are small-ruminant betaretroviruses that share high nucleotide and amino acid identity, utilize the same cellular receptor, hyaluronoglucosaminidase 2 (Hyal2) for entry, and transform tissues with their envelope (Env) glycoprotein; yet, they target discrete regions of the respiratory tract-the lung and nose, respectively. This distinct tissue selectivity makes them ideal tools with which to study the pathogenesis of betaretroviruses. To uncover the genetic determinants of tropism, we constructed JSRV-ENTV chimeric viruses and produced lentivectors pseudotyped with the Env proteins from JSRV (Jenv) and ENTV (Eenv). Through the transduction and infection of lung and nasal turbinate tissue slices, we observed that Hyal2 expression levels strongly influence ENTV entry, but that the long terminal repeat (LTR) promoters of these viruses are likely responsible for tissue-specificity. Furthermore, we show evidence of ENTV Env expression in chondrocytes within ENTV-infected nasal turbinate tissue, where Hyal2 is highly expressed. Our work suggests that the unique tissue tropism of JSRV and ENTV stems from the combined effort of the envelope glycoprotein-receptor interactions and the LTR and provides new insight into the pathogenesis of ENTV.


Subject(s)
Gene Products, env/genetics , Jaagsiekte sheep retrovirus/physiology , Oncogenic Viruses/physiology , Pulmonary Adenomatosis, Ovine/virology , Terminal Repeat Sequences , Tumor Virus Infections/virology , Viral Tropism , Animals , Cell Line , Gene Order , Genome, Viral , Host Specificity , Host-Pathogen Interactions , Humans , Reassortant Viruses/genetics , Sheep
11.
J Virol ; 93(21)2019 11 01.
Article in English | MEDLINE | ID: mdl-31434729

ABSTRACT

Jaagsiekte sheep retrovirus (JSRV) is the etiologic agent of ovine pulmonary adenocarcinoma (OPA), a neoplastic lung disease of sheep. OPA is an important economic and welfare issue for sheep farmers and a valuable naturally occurring animal model for human lung adenocarcinoma. Here, we used RNA sequencing to study the transcriptional response of ovine lung tissue to infection by JSRV. We identified 1,971 ovine genes differentially expressed in JSRV-infected lung compared to noninfected lung, including many genes with roles in carcinogenesis and immunomodulation. The differential expression of selected genes was confirmed using immunohistochemistry and reverse transcription-quantitative PCR. A key finding was the activation of anterior gradient 2, yes-associated protein 1, and amphiregulin in OPA tumor cells, indicating a role for this oncogenic pathway in OPA. In addition, there was differential expression of genes related to innate immunity, including genes encoding cytokines, chemokines, and complement system proteins. In contrast, there was little evidence for the upregulation of genes involved in T-cell immunity. Many genes related to macrophage function were also differentially expressed, reflecting the increased abundance of these cells in OPA-affected lung tissue. Comparison of the genes differentially regulated in OPA with the transcriptional changes occurring in human lung cancer revealed important similarities and differences between OPA and human lung adenocarcinoma. This study provides valuable new information on the pathogenesis of OPA and strengthens the use of this naturally occurring animal model for human lung adenocarcinoma.IMPORTANCE Ovine pulmonary adenocarcinoma is a chronic respiratory disease of sheep caused by jaagsiekte sheep retrovirus (JSRV). OPA is a significant economic problem for sheep farmers in many countries and is a valuable animal model for some forms of human lung cancer. Here, we examined the changes in host gene expression that occur in the lung in response to JSRV infection. We identified a large number of genes with altered expression in infected lung, including factors with roles in cancer and immune system function. We also compared the data from OPA to previously published data from human lung adenocarcinoma and found a large degree of overlap in the genes that were dysregulated. The results of this study provide exciting new avenues for future studies of OPA and may have comparative relevance for understanding human lung cancer.


Subject(s)
Jaagsiekte sheep retrovirus/physiology , Lung/virology , Pulmonary Adenomatosis, Ovine/genetics , Adenocarcinoma of Lung/genetics , Animals , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Host-Pathogen Interactions , Humans , Lung/metabolism , Lung/pathology , Lung Neoplasms/genetics , Pulmonary Adenomatosis, Ovine/metabolism , Pulmonary Adenomatosis, Ovine/pathology , Pulmonary Adenomatosis, Ovine/virology , Sheep
12.
Vet Res ; 48(1): 76, 2017 11 14.
Article in English | MEDLINE | ID: mdl-29137669

ABSTRACT

JSRV (Jaagsiekte Sheep Retrovirus) is a retrovirus inducing a transmissible lung adenocarcinoma in sheep and goats with predominantly lepidic and papillary lesions. This naturally occurring lung cancer in large animals shares many features with human pneumonic-type lung adenocarcinomas with predominant lepidic growth. The metastatic spread is rare in both human and animal cancers. This unique feature prompted us to decipher the angiogenesis pathway in these cancers. We focused on the levels of mRNA and proteins of genes implicated in the extension of JSRV-induced lung adenocarcinomas by studying their expression in lung cancers (n = 10) and normal lungs (n = 10) and in primary epithelial alveolar type II cells derived from cancers (n = 10) or normal lungs (n = 6). In parallel, we evaluated the levels of expression of key genes in lung tissues collected from lepidic (n = 13) or papillary (n = 5) human adenocarcinomas and, when available, adjacent normal lungs (n = 11). We measured the expression of the same key genes implicated in angiogenesis, lymphangiogenesis and degradation of the extracellular matrix. In ovine adenocarcinomas, VEGFR2 and VEGFD mRNA were downregulated in cancers; MMP9, TIMP1 and FGFR2 mRNA were overexpressed as compared to normal lungs. Importantly, VEGFA and VEGFR2 proteins were not expressed in JSRV-induced cancers. In human lepidic adenocarcinomas, VEGFA and VEGFR2 mRNA were weakly expressed and no VEGFR2 protein was detectable. Downregulation of key angiogenic players may contribute to the control of extra thoracic invasion of cancer cells in human and ovine pneumonic-type adenocarcinoma with predominant lepidic growth.


Subject(s)
Adenocarcinoma/genetics , Gene Expression Regulation, Neoplastic , Jaagsiekte sheep retrovirus/physiology , Lung Neoplasms/genetics , Neovascularization, Pathologic/genetics , Neovascularization, Pathologic/veterinary , Pulmonary Adenomatosis, Ovine/genetics , Adenocarcinoma/metabolism , Adenocarcinoma of Lung , Adenocarcinoma, Papillary/genetics , Adenocarcinoma, Papillary/metabolism , Adult , Aged , Aged, 80 and over , Animals , Female , Humans , Lung/physiopathology , Lung Neoplasms/metabolism , Male , Middle Aged , Pulmonary Adenomatosis, Ovine/metabolism , Sheep
13.
Res Vet Sci ; 114: 332-340, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28651195

ABSTRACT

Surfactant covers the inner surface of lung alveoli and lowers the surface tension to prevent alveoli from collapsing. A lack of surfactant or its dysfunction causes dyspnea. The Jaagsiekte Sheep Retrovirus (JSRV) causes ovine pulmonary adenocarcinoma (OPA), whose typical clinical appearance is fluid running from nostrils. This fluid might contain surfactant as alveolar type II pneumocytes (AEII) are target cells for JSRV. Therefore, the progressive dyspnea during OPA might be caused partially by surfactant alterations. Bronchoalveolar and intracellular surfactant as well as the biophysical function of surfactant were analyzed in OPA sheep and controls. Transmission electron microscopy and stereological methods were used to characterize ultrastructure and distribution of surfactant subtypes in AEII and bronchoalveolar lavage fluid (BALF). Pulsating Bubble Surfactometry enabled studying the surface activity of the surfactant, while lung volumes were detected by computed tomography. The methods used are suitable to determine intraalveolar and intracellular surfactant subtypes in OPA sheep and controls. OPA sheep showed more lamellar body-like forms, multivesicular vesicles and tubular myelin in BALF compared to controls. These higher amounts of active surfactant subtypes might be a consequence of a higher surfactant production and release. Surfactant subtypes in AEII of OPA sheep showed smaller and more immature lamellar bodies compared to controls. The surfactant surface activity of OPA sheep does not show obvious defects. In conclusion, the general quality of surfactant in OPA appears to be equivalent to surfactant produced in controls, however, dyspnea of OPA might be triggered by quantity of fluid production.


Subject(s)
Adenocarcinoma/physiopathology , Pulmonary Adenomatosis, Ovine/physiopathology , Pulmonary Alveoli/physiopathology , Animals , Jaagsiekte sheep retrovirus/physiology , Reference Values , Sheep
14.
PLoS One ; 12(4): e0175586, 2017.
Article in English | MEDLINE | ID: mdl-28419107

ABSTRACT

Developing lung cancer in mouse models that display similarities of both phenotype and genotype will undoubtedly provide further and better insights into lung tumor biology. Moreover, a high degree of pathophysiological similarity between lung tumors from mouse models and their human counterparts will make it possible to use these mouse models for preclinical tests. Ovine pulmonary adenocarcinomas (OPAs) present the same symptoms as adenocarcinomas in humans and are caused by a betaretrovirus. OPAs have served as an exquisite model of carcinogenesis for human lung adenocarcinomas. In this study, we characterized the histopathology and transcriptome profiles of a jaagsiekte sheep retrovirus (JSRV)-envelope protein (Env) transgenic mouse model with spontaneous lung tumors, and associations of the transcriptome profiles with tumor invasion/metastasis, especially the phenomenon of the epithelial-mesenchymal transition (EMT). Genetic information obtained from an expression array was analyzed using an ingenuity pathways analysis (IPA) and human disease database (MalaCards). By careful examination, several novel EMT-related genes were identified from tumor cells using RT-qPCR, and these genes also scored high in MalaCards. We concluded that the JSRV-Env mouse model could serve as a spontaneous lung adenocarcinoma model with a metastatic phenotype, which will benefit the study of early-onset and progression of lung adenocarcinoma. In addition, it can also be a valuable tool for biomarkers and drug screening, which will be helpful in developing intervention therapies.


Subject(s)
Adenocarcinoma/pathology , Disease Models, Animal , Lung Neoplasms/pathology , Lung/pathology , Adenocarcinoma/genetics , Adenocarcinoma/metabolism , Animals , Epithelial-Mesenchymal Transition/genetics , Gene Expression Profiling/methods , Gene Expression Regulation, Neoplastic , Humans , Immunohistochemistry , Jaagsiekte sheep retrovirus/genetics , Lung/metabolism , Lung Neoplasms/genetics , Lung Neoplasms/metabolism , Mice, Transgenic , Multidrug Resistance-Associated Proteins/analysis , Neoplasm Metastasis , Nuclear Proteins/analysis , Phenotype , Pulmonary Adenomatosis, Ovine/genetics , Reverse Transcriptase Polymerase Chain Reaction , Sheep , Thyroid Nuclear Factor 1 , Transcription Factors/analysis , Viral Envelope Proteins/genetics
15.
Biochem Biophys Res Commun ; 485(3): 672-678, 2017 Apr 08.
Article in English | MEDLINE | ID: mdl-28235485

ABSTRACT

The envelope (Env) of Jaagsiekte sheep retrovirus (JSRV) is an oncoprotein of ovine pulmonary adenocarcinoma (OPA). Autophagy is involved in different cancers, but how it is carcinogenic in JSRV Env is unclear. Modulation of autophagy in exJSRV-env-NM-transfected cells through the Akt/mTOR and MAPK signaling pathway was studied, and we observed strong positive labeling of p-Akt, p-mTOR, p-MEK1/2, p-ERK1/2, p-p38 and p-JNK in tumor cells and typical type II pneumocytes in naturally infected OPA lung tissues, which was co-aligned with JSRV-Env positive cells as shown by immunohistochemical and microscopic analysis. Akt/mTOR and MAPK pathways were activated in OPA lung and JSRV-Env transfected NIH 3T3 cells. Decreased Beclin1 and LC3 II/I suggested that autophagy was inhibited in OPA lung and JSRV-Env transfected NIH 3T3 cells. Beclin1 and LC3 II/I increased in JSRV-Env transfected NIH3T3 cells treated with mTOR inhibitor (rapamycin), ERK1/2 inhibitor (PD 98059), p38 inhibitor (SB 203580) and JNK inhibitor (SP 600125), suggesting that Akt/mTOR and MAPK pathways were responsible for JSRV-Env decreased autophagy. In conclusion, JSRV Env decreased autophagy in JSRV-Env transfected NIH3T3 cells through Akt/mTOR and MAPK pathways, in particular, JNK and p38 pathways.


Subject(s)
Autophagy , Gene Products, env/metabolism , Jaagsiekte sheep retrovirus/metabolism , MAP Kinase Signaling System , Proto-Oncogene Proteins c-akt/metabolism , Pulmonary Adenomatosis, Ovine/metabolism , TOR Serine-Threonine Kinases/metabolism , Animals , Extracellular Signal-Regulated MAP Kinases/metabolism , Fibroblasts/metabolism , Fibroblasts/virology , Gene Products, env/genetics , Host-Pathogen Interactions , Immunoblotting , Immunohistochemistry , JNK Mitogen-Activated Protein Kinases/metabolism , Jaagsiekte sheep retrovirus/genetics , Jaagsiekte sheep retrovirus/physiology , Lung/metabolism , Lung/virology , Mice , NIH 3T3 Cells , Phosphorylation , Pulmonary Adenomatosis, Ovine/genetics , Pulmonary Adenomatosis, Ovine/virology , Sheep , p38 Mitogen-Activated Protein Kinases/metabolism
16.
Genet Mol Res ; 15(3)2016 Aug 29.
Article in English | MEDLINE | ID: mdl-27706644

ABSTRACT

The envelope protein (Env) of the Jaagsiekte sheep retrovirus (JSRV) is known to be a unique oncoprotein responsible for inducing ovine pulmonary adenocarcinoma (OPA). The objective of this study was to prepare a specific monoclonal antibody (mAb) against the JSRV Env protein using bioinformatic analysis. According to the structure and epitope prediction results of JSRV Env, the JSRV-Env572-615 antigen was prepared via peptide synthesis (amino acid sequence 572-615, denoted as JSRV-Env572-615). BALB/c mice were immunized to prepare the anti-JSRV-Env572-615 mAb. Spleen cells were fused with SP2/0 myeloma cells after being screened by indirect ELISA and cloned by limiting dilution. The specificity of mAb was evaluated by western blot analysis and immunohistochemistry assays. Western blot results showed that the JSRV Env protein was able to bind to mAb with high specificity. Immunohistochemistry assays demonstrated that the mAb was able to recognize JSRV Env in adenomatous hyperplasia of the lung. Furthermore, JSRV was detected in peripheral blood leukocytes during the pre-clinical period of OPA in 2 of the 25 sheep using this newly synthesized mAb. Therefore, this mAb may be a useful tool for the detection of JSRV in sheep.


Subject(s)
Adenocarcinoma/diagnosis , Adenocarcinoma/veterinary , Antibodies, Monoclonal/biosynthesis , Antibodies, Viral/biosynthesis , Jaagsiekte sheep retrovirus/immunology , Lung Neoplasms/diagnosis , Lung Neoplasms/veterinary , Pulmonary Adenomatosis, Ovine/diagnosis , Adenocarcinoma/immunology , Adenocarcinoma/virology , Adenocarcinoma of Lung , Amino Acid Sequence , Animals , Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/isolation & purification , Antibodies, Viral/chemistry , Antibodies, Viral/isolation & purification , Antibody Specificity , Computational Biology , Early Diagnosis , Epitopes/chemistry , Epitopes/immunology , Gene Products, env/chemistry , Gene Products, env/immunology , Jaagsiekte sheep retrovirus/isolation & purification , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/virology , Lung/immunology , Lung/virology , Lung Neoplasms/immunology , Lung Neoplasms/virology , Mice , Mice, Inbred BALB C , Peptides/administration & dosage , Peptides/chemical synthesis , Peptides/immunology , Pulmonary Adenomatosis, Ovine/immunology , Pulmonary Adenomatosis, Ovine/virology , Sheep , Sheep, Domestic , Spleen/cytology , Spleen/immunology
17.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 32(9): 1188-92, 2016 Sep.
Article in Chinese | MEDLINE | ID: mdl-27609573

ABSTRACT

Objective To explore the influence of the exogenous Jaagsiekte sheep retrovious (exJSRV) envelope protein (Env) on NIH3T3 cell proliferation. Methods A recombinant plasmid pcDNA4/myc-His/exJSRV- env carrying exJSRV- env gene was constructed, and then the correctness of the recombinant plasmid was identified by PCR, restriction enzyme digestion and sequencing. The recombinant plasmid pcDNA4/myc-His/exJSRV- env was transiently transfected into NIH3T3 cells by Lipofectamine(TM) LTX. After the transfection of the recombinant plasmid, the expression of exJSRV- env was detected by reverse transcription PCR and Western blotting. The effect of Env on cell proliferation was investigated by CCK-8 assay and plate colony formation assay. Results The recombinant eukaryotic expression plasmid containing exJSRV- env was successfully constructed as identified by PCR, restriction enzyme identification and sequencing. After the recombinant plasmid was transiently transfected into NIH3T3 cells, reverse transcription PCR and Western blotting showed the expression of exJSRV- env , and Env promoted NIH3T3 cell proliferation significantly. Conclusion JSRV Env was expressed successfully in the NIH3T3 cells and promoted the proliferation of NIH3T3 cells.


Subject(s)
Betaretrovirus/genetics , Cell Proliferation , Pulmonary Adenomatosis, Ovine/physiopathology , Pulmonary Adenomatosis, Ovine/virology , Viral Envelope Proteins/genetics , Animals , Betaretrovirus/metabolism , Gene Expression , Mice , NIH 3T3 Cells , Sheep , Transfection , Viral Envelope Proteins/metabolism
19.
Vet Pathol ; 53(6): 1172-1179, 2016 11.
Article in English | MEDLINE | ID: mdl-27154541

ABSTRACT

Ovine pulmonary adenocarcinoma (OPA) is a contagious lung cancer of sheep caused by jaagsiekte sheep retrovirus (JSRV). It is generally accepted that transmission by the respiratory route occurs under natural conditions. However recent studies strongly indicate that JSRV can also be transmitted to lambs perinatally via colostrum and milk (C/M). The aim of this work was to confirm that C/M can transmit JSRV infection to lambs under natural conditions and investigate the initial events associated with this transmission route. We have analyzed the presence of JSRV in C/M samples from 22 naturally infected, asymptomatic ewes throughout a lactation period, and in various tissues collected from a group of 36 of their lambs that were fed naturally. The lambs were euthanized at 12, 24, 48, and 72 hours and at 5 and 10 days after birth. We detected JSRV-provirus by PCR in the somatic C/M cells from 10/22 ewes (45.45%). The virus was also detected in 9/36 lambs (25%). JSRV-infected cells, with lymphoreticular-like morphology, were observed by immunohistochemistry (IHC) and in situ hybridization (ISH) in Peyer's patches (PP) from the small intestine of the youngest lambs and in mesenteric lymph nodes (MLN) from lambs older than 72 hours. The virus was also detected by PCR in white blood cells (WBC) in 2/36 lambs (5.5%). These results confirm colostral transmission of JSRV to lambs under natural conditions. Infected lymphoreticular cells contained in C/M appear to be involved. These cells can cross the intestinal barrier of newborn lambs, reach the MLN and enter into circulation.


Subject(s)
Infectious Disease Transmission, Vertical/veterinary , Jaagsiekte sheep retrovirus , Lymph Nodes/virology , Peyer's Patches/virology , Pulmonary Adenomatosis, Ovine/transmission , Animals , Animals, Suckling/virology , Female , Male , Mesentery , Milk/virology , Polymerase Chain Reaction/veterinary , Sheep
20.
J Vet Diagn Invest ; 28(3): 249-56, 2016 May.
Article in English | MEDLINE | ID: mdl-27016721

ABSTRACT

Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring cancer in sheep that is caused by the Jaagsiekte sheep retrovirus (JSRV). Because the pathologic and epidemiologic features of OPA are similar to those of bronchoalveolar carcinoma in humans, OPA is considered a useful animal model for pulmonary carcinogenesis. In this study, 3,512 lungs from various breeds of sheep were collected and macroscopically examined. OPA was identified in 30 sheep, and samples of these animals were further examined by histologic, immunohistochemical (p53 protein, surfactant protein A [SP-A], proliferating cell nuclear antigen [PCNA], JSRV matrix protein [MA]), and PCR methods. Papillary or acinar adenocarcinomas were detected microscopically in the affected areas. Immunoreactivity for p53 PAb240 was detected in 13 sheep, whereas p53 DO-1 was not detected in any of the OPA animals. PCNA immunoreactivity was recorded in 27 animals. SP-A and JSRV MA protein was immunopositive in all 30. JSRV proviral DNA was detected by PCR analysis in all of the lung samples collected from OPA animals. In addition, the pulmonary SP-A levels were increased in tumor cells. The results of this study suggest that PCNA and p53 protein expression may be useful indicators in monitoring malignancy of pulmonary tumors.


Subject(s)
Pulmonary Adenomatosis, Ovine/virology , Animals , Immunohistochemistry/veterinary , Jaagsiekte sheep retrovirus/pathogenicity , Lung/pathology , Polymerase Chain Reaction/veterinary , Pulmonary Adenomatosis, Ovine/metabolism , Pulmonary Adenomatosis, Ovine/pathology , Pulmonary Surfactant-Associated Protein A/metabolism , Sheep , Tumor Suppressor Protein p53/metabolism
SELECTION OF CITATIONS
SEARCH DETAIL
...