Cases Of Ovine Pulmonary Adenocarcinoma Research Articles

Retroviral coinfection (Jaagsiekte and Maedi-Visna viruses) in sheep with pulmonary tumors in Transylvania (Romania): retrospective study on 82 cases.

Ovine pulmonary adenocarcinoma (OPA) is an important viral-induced neoplasia in sheep caused by exogenous Jaagsiekte sheep retrovirus (exJSRV). Coinfection of exJSRV and Maedi-Visna virus (MVV) is reported in OPA cases, but its worldwide distribution and significance on lung pathology is not yet completely understood. This study aimed to investigate the MVV coinfection rate in 82 exJSRV-related OPA cases, and their pathological effects on lung parenchyma in slaughtered sheep in Transylvania (Romania). On gross examination, classical form of OPA was identified in 92.7%; no changes consisting with MVV interstitial pneumonia were identified in the included cases. The most common histological type of OPA was acinar (58.5%) and the myxoid growths were found in 18 cases. The exJSRV and MMV coinfection rate in examined sheep was 47.6% (39/82). The assessment of perineoplastic areas from coinfected animals, revealed interstitial lymphoplasmacytic infiltrates in all cases, lymphoid hyperplasia in 60.6% cases (20/33) and fibromuscular hyperplasia in 63.7% (21/33). This is the first report providing new data on distribution of OPA coexisting with MVV infection in slaughtered sheep in Romania. We consider that the OPA and MVV coinfection may play an important role on the severity of ovine chronic pulmonary diseases and further studies are needed to confirm this hypothesis.

A Combined Approach for Detection of Ovine Small Ruminant Retrovirus Co-Infections.

Jaagsiekte retrovirus (JSRV)-induced ovine pulmonary adenocarcinoma (OPA) is an important ovine respiratory disease in Switzerland. Furthermore, ovine lungs with OPA frequently exhibited lesions suggestive of maedi-visna virus (MVV) or caprine arthritis encephalitis virus (CAEV) infection, indicating that co-morbidities might occur. Lungs and pulmonary lymph nodes were sampled from suspected OPA cases, inflammatory lung lesions and control lungs (total of 110 cases). Tissues were (a) processed for histology and immunohistochemistry (IHC), and (b) underwent DNA extraction and real-time PCR for JSRV, MVV and CAEV. Peptide sequences were used to generate virus-specific customized polyclonal antibodies. PCR-positive OPA cases and formalin-fixed and paraffin-embedded MVV- and CAEV-infected synovial cell pellets served as positive controls. Fifty-two lungs were histologically diagnosed with OPA. Histological evidence of MVV/CAEV infection was detected in 25 lungs. JSRV was detected by PCR in 84% of the suspected OPA cases; six were co-infected with MVV and one with CAEV. MVV was detected by PCR in 14 cases, and four lungs were positive for CAEV. Three lungs had MVV/CAEV co-infection. In IHC, JSRV was detected in 91% of the PCR-positive cases, whereas MVV and CAEV immunoreactivity was seen in all PCR-positive lungs. Although PCR showed a higher sensitivity compared to IHC, the combined approach allows for investigations on viral cell tropism and pathogenic processes in co-morbidities, including their potential interdependency. Furthermore, an immunohistochemical tool for specific differentiation of MVV and/or CAEV infection was implemented.

Clinical, molecular, and pathological investigations of ovine pulmonary adenocarcinoma in the middle of Iraq.

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.

Exogenous Small Ruminant Betaretrovirus Envelope Protein Is Detected in Draining Lymph Nodes in Contagious Respiratory Tumors of Sheep and Goats.

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.

A Novel Translational Ovine Pulmonary Adenocarcinoma Model for Human Lung Cancer.

In vitro cell line and in vivo murine models have historically dominated pre-clinical cancer research. These models can be expensive and time consuming and lead to only a small percentage of anti-cancer drugs gaining a license for human use. Large animal models that reflect human disease have high translational value; these can be used to overcome current pre-clinical research limitations through the integration of drug development techniques with surgical procedures and anesthetic protocols, along with emerging fields such as implantable medical devices. Ovine pulmonary adenocarcinoma (OPA) is a naturally-occurring lung cancer that is caused by the jaagsiekte sheep retrovirus. The disease has similar histological classification and oncogenic pathway activation to that of human lung adenocarcinomas making it a valuable model for studying human lung cancer. Developing OPA models to include techniques used in the treatment of human lung cancer would enhance its translational potential, making it an excellent research tool in assessing cancer therapeutics. In this study we developed a novel OPA model to validate the ability of miniaturized implantable O2 and pH sensors to monitor the tumor microenvironment. Naturally-occurring pre-clinical OPA cases were obtained through an on-farm ultrasound screening programme. Sensors were implanted into OPA tumors of anesthetized sheep using a CT-guided trans-thoracic percutaneous implantation procedure. This study reports the findings from 9 sheep that received sensor implantations. Time taken from initial CT scans to the placement of a single sensor into an OPA tumor was 45 ± 5 min, with all implantations resulting in the successful delivery of sensors into tumors. Immediate post-implantation mild pneumothoraces occurred in 4 sheep, which was successfully managed in all cases. This is, to the best of our knowledge, the first description of the use of naturally-occurring OPA cases as a pre-clinical surgical model. Through the integration of techniques used in the treatment of human lung cancer patients, including ultrasound, general anesthesia, CT and surgery into the OPA model, we have demonstrated its translational potential. Although our research was tailored specifically for the implantation of sensors into lung tumors, we believe the model could also be developed for other pre-clinical applications.

Differential immunohistochemical expression of JSRV capsid antigen and tumour biomarkers in classical and atypical OPA: a comparative study

ABSTRACT Ovine pulmonary adenocarcinoma (OPA) is a retroviral disease of sheep and occurs usually in two forms viz. classical and atypical, which differ in their histological lesions, cellular infiltrates, behaviour and prognosis. Present investigation was focussed to evaluate the differential expression of various tumour biomarkers in both OPA forms. Among 23 lung tissue samples suspected for OPA, histopathological examination diagnosed 12 as classical OPA, 6 as atypical OPA and 5 as bronchiolar hyperplasia. Polymerase chain reaction with LTR (long terminal repeat)-specific primers revealed that all classical and atypical OPA cases and three bronchiolar hyperplasia cases were positive for Jaagsiekte sheep retrovirus (JSRV) genome. Immunohistochemical (IHC) staining showed that JSRV capsid antigen was localized in the proliferating alveolar pneumocytes, bronchiolar epithelium and MNCs (mononuclear cells). Expression of PCNA was comparatively higher in classical OPA than atypical OPA and bronchiolar hyperplasia. Atypical OPA had higher MHC II expression than classical form indicated the higher proliferation of fibroblasts. FOXO3a expression is comparatively higher in OPA cases with mild inflammatory reaction while MMP2 expression was higher in OPA cases with severe inflammatory reaction. In conclusion, classical OPA is more proliferative and expansive in nature as compared to the atypical form. Further controlled study is required to prove the prognostic and metastatic potential of these markers in OPA.

Ovine Pulmonary Adenocarcinoma: A Unique Model to Improve Lung Cancer Research.

Lung cancer represents a major worldwide health concern; although advances in patient management have improved outcomes for some patients, overall 5-year survival rates are only around 15%. In vitro studies and mouse models are commonly used to study lung cancer and their use has increased the molecular understanding of the disease. Unfortunately, mouse models are poor predictors of clinical outcome and seldom mimic advanced stages of the human disease. Animal models that more accurately reflect human disease are required for progress to be made in improving treatment outcomes and prognosis. Similarities in pulmonary anatomy and physiology potentially make sheep better models for studying human lung function and disease. Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring lung cancer that is caused by the jaagsiekte sheep retrovirus. The disease is endemic in many countries throughout the world and has several features in common with human lung adenocarcinomas, including histological classification and activation of common cellular signaling pathways. Here we discuss the in vivo and in vitro OPA models that are currently available and describe the advantages of using pre-clinical naturally occurring OPA cases as a translational animal model for human lung adenocarcinoma. The challenges and options for obtaining these OPA cases for research purposes, along with their use in developing novel techniques for the evaluation of chemotherapeutic agents or for monitoring the tumor microenvironment in response to treatment, are also discussed.

Diagnosis and prevalence of ovine pulmonary adenocarcinoma in lung tissues of naturally infected farm sheep

Aim:This study was aimed to detect ovine pulmonary adenocarcinoma (OPA) in sheep flocks affected with pulmonary disorders at organized farm.Materials and Methods:A total of 75 sheep died naturally were thoroughly examined for the lesions of OPA during necropsy. Tissue sections from affected portion of the lungs from each animal were collected aseptically and divided into two parts; one each for polymerase chain reaction (PCR) and another for histopathology.Results:On PCR examination of lung tissues, six sheep (8%) were found to be positive for JSRV. Two of them were 3-6 months of age and did not show clinical signs/gross lesions of OPA. Four adult sheep positive on PCR revealed characteristic lesions of OPA on gross and histopathological examination.Conclusion:In the absence of known specific antibody response to the infection with JSRV, there is no diagnostic serological test available. The PCR assay employed in this study on lung tissues, using primers based on the U3 region of the viral long terminal repeat for JSRV would be helpful in the screening of preclinical and clinical cases of OPA in sheep.

Detection of exogenous Jaagsiekte sheep retrovirus in Turkey

The aim of this study is to detect the exogenous Jaagsiekte sheep retrovirus (exJSRV) in suspected cases of ovine pulmonary adenocarcinoma (OPA) from the Eastern Anatolian region of Turkey. Pathological examination and PCR were carried out with the lung, lymph nodule, brain, heart and liver tissues of four sheep with suspected OPA. Histology of the lung sections indicated the well-circumscribed, multifocal and unencapsulated gray to white masses (arrows) on the parietal surface of medial and caudal lobes. exJSRV was detected in all tissue samples except for brain by nested polymerase chain reaction (nPCR). In addition to the nested-PCR results, the presence of exJSRV into the clinical samples was confirmed with sequencing of two PCR-positive products for OPAV. This report highlights the first presence of exJSRV in the sheep suspected with OPA in Turkey. Furthermore, the results provide supporting evidence for the metastasis of exJSRV in extra-thoracic tissues.

Antibiotic Treatment Response of Chronic Lung Diseases of Adult Sheep in the United Kingdom Based upon Ultrasonographic Findings.

Examination of the lungs of adult sheep with chronic respiratory diseases was readily achieved using both 5 MHz linear and sector scanners. Superficial lung abscesses in eight sheep appeared as anechoic areas containing multiple hyperechoic dots bordered distally by a broad hyperechoic capsule. Unilateral fibrinous pleurisy (2 sheep) appeared as an anechoic area containing a hyperechoic latticework. Ovine pulmonary adenocarcinoma (OPA) lesions appeared as sharply demarcated hypoechoic areas in the lung parenchyma initially in the cranioventral lung lobes (21 sheep) with lesions also present in the caudodorsal diaphragmatic lobe (11 sheep); abscesses and areas of calcification within the OPA tumour mass were also identified. Daily treatment with procaine penicillin for 30 consecutive days was successful in both sheep with unilateral fibrinous pleurisy and six sheep identified with superficial lung abscesses measuring 2–8 cm in diameter; only one of two sheep with more extensive lesions recovered. Auscultation of the chest failed to detect adventitious sounds in any of the ten sheep with lung abscesses; normal breath sounds were reduced over the area of fibrinous pleurisy; no pleuritic rubs were heard. Wheezes and crackles auscultated in some OPA cases and did not correlate well with lesions detected ultrasonographically.

Nucleotide Sequencing and Phylogenetic Analysis Using PCR Amplicons of U3 Gene of Jaagsiekte Sheep Retrovirus (JSRV) Detected in Natural Cases of Ovine Pulmonary Adenocarcinoma in India

Ovine pulmonary adenocarcinoma (OPA) caused by an exogenous Jaagsiekte sheep retrovirus (JSRV) is prevalent in Indian sheep. In the present study, OPA was diagnosed in sheep by clinical signs, gross and histopathology and polymerase chain reaction (PCR). Proviral DNA of exogenous JSRV was detected in lung tumor tissues, mediastinal lymph nodes, blood and lung fluid samples from natural cases of OPA by using U3-hn PCR and the PCR amplicons were sequenced to analyze nucleotide divergence. In total, six isolates were sequenced that had 96% - 100% homology with a UK strain (AF105220.1) but more divergent from a South African strain (M80216) with 88% - 93% identity. The phylogenetic analysis revealed segregation of the six isolates into two clusters. In conclusion, this study is the first report on sequencing and phylogenetic analysis of JSRV in India and further studies are suggested to know the complete sequencing and genetic divergence of JSRV in Indian sheep.

Lung auscultation recordings from normal sheep and from sheep with well-defined respiratory tract pathology

Increased audibility of normal lung sounds is commonly caused by hyperventilation after exercise and during hot weather. Tachypnoea is common in toxaemic and septicaemic conditions, but there may be no adventitious sounds. Moderate to pronounced coarse crackles are readily identified in advanced cases of ovine pulmonary adenocarcinoma, but auscultation findings do not correspond well to the distribution of lesion(s) revealed at necropsy. Auscultation does not detect pleural abscesses (up to 10 cm diameter). Marked attenuation of normal lung sounds results from marked fibrinous pleurisy and extensive unilateral pyothorax. Pleural frictions rubs are not heard in cases of marked fibrinous pleurisy associated with pleural abscesses. Rumen contraction sounds are often superimposed upon lung sounds. Auscultation of the ovine chest alone does not allow the clinician to determine the presence of all superficial lung pathology nor accurately define its distribution. Ultrasonography provides more accurate information regarding the nature and extent of superficial lung pathology in sheep.

Lung Tumor Development and Spontaneous Regression in Lambs Coinfected With Jaagsiekte Sheep Retrovirus and Ovine Lentivirus

Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring and experimentally inducible lung cancer of sheep caused by Jaagsiekte sheep retrovirus (JSRV). The first aim of this study was to monitor the development of OPA with minimally invasive, real-time observations of animals experimentally infected with JSRV as well as ovine lentivirus (maedi-visna virus). Worldwide, simultaneous infection of sheep with these 2 retroviruses is a common occurrence, naturally and experimentally; consequently, the lung tumor homogenates used as inocula contained both viruses. Following inoculation, computed tomography was used to detect tumor nodules early, before the onset of clinical signs, and to monitor tumor advancement. However, not only was OPA disease progression observed, but the apparent spontaneous regression of OPA was witnessed. In fact, regression was more common than progression following JSRV inoculation of neonatal lambs. Immune responses were detected, particularly involving CD3(+) T cells and the production of antibodies against JSRV that may mediate the spontaneous regression of JSRV-induced OPA. The second aim of this study was to determine whether OPA tumors harbor genetic alterations similar to those found in human lung adenocarcinoma. No mutations were found in the tyrosine kinase domain of the epidermal growth factor receptor, KRAS codons 12 and 13, or the DNA-binding domain of p53 in tumor DNA from naturally occurring and experimentally-induced OPA cases. Overall, the genetic profile combined with the disease development data provides further important characterization of OPA and describes, for the first time, spontaneous regression of OPA tumors in experimentally infected sheep.

Relationship between thoracic auscultation and lung pathology detected by ultrasonography in sheep

The utility of routine auscultation to detect and characterise the nature of a range of superficial lung and pleural pathologies in domestic sheep was assessed using ultrasonographic examination to indicate and localise pathologies pre-mortem. Necropsy examination was then used to fully characterise the nature and extent of the lesions. Auscultation recordings were made from 10 normal sheep with no clinical evidence of respiratory disease and with absence of significant superficial lung pathology, which was confirmed initially by ultrasound examination and subsequently at necropsy examination. A further two sheep with endotoxaemia and 30 sheep with well-defined lung lesions were also examined. Increased audibility of normal lung sounds in 4/10 normal sheep was associated with tachypnoea as a consequence of handling and transport during hot weather and was also observed in the two sheep with endotoxaemia. Moderate to severe coarse crackles detected in all advanced cases of ovine pulmonary adenocarcinoma (n=16) were audible over an area larger than the lesion distribution identified during ultrasound examination, and confirmed later at necropsy. Auscultation did not detect abnormal sounds in any of the five sheep with focal pleural abscesses (up to 10 cm diameter). Unilateral pyothorax caused attenuation of sounds relative to the contra-lateral normal lung in all three sheep with this condition. Marked fibrinous pleurisy caused attenuation of sounds relative to normal areas of lung in six sheep. No sounds resembling the description of pleural frictions rubs were heard in the sheep with marked fibrinous pleurisy (n=6) or associated with focal pleural abscesses (n=5). Routine interpretation of auscultated sound did not allow the presence of superficial lung pathology or its distribution to be accurately defined in the respiratory diseases represented in this study.

Jaagsiekte sheep retrovirus is present at high concentration in lung fluid produced by ovine pulmonary adenocarcinoma-affected sheep and can survive for several weeks at ambient temperatures

Jaagsiekte sheep retrovirus (JSRV) causes a fatal lung cancer of sheep known as ovine pulmonary adenocarcinoma (OPA). OPA is a significant disease in many sheep-rearing countries and there is no effective method of control. A unique feature of OPA is the overproduction of fluid in the lung of affected animals. This lung fluid contains JSRV and provides a means of transmission through the inhalation of virus. In this study we demonstrated that lung fluid from different OPA cases contained between 10 7 and 10 10 copies of JSRV RNA per ml. Examination of JSRV RNA survival under conditions that mimic natural conditions suggested that intact JSRV virions may persist for several weeks in the environment. These are the first quantitative data on JSRV in lung fluid and provide valuable information for implementing appropriate biosecurity measures to control the spread of JSRV in the field.

In-situ Demonstration of Mitogen-activated Protein Kinase Erk 1/2 Signalling Pathway in Contagious Respiratory Tumours of Sheep and Goats

Ovine pulmonary adenocarcinoma (OPA) and enzootic nasal adenocarcinoma (ENA) are two contagious neoplastic diseases of secretory epithelial cells in the respiratory system of sheep and goats. Jaagsiekte sheep retrovirus (JSRV) is the aetiological agent of OPA, and enzootic nasal tumour virus (ENTV) is associated with ENA. The genomes of these retroviruses do not contain known oncogenes but products of the env gene are important in the generation of transforming stimuli. However, the cell signalling pathways activated in vivo are not completely understood. This study was based on the use of activation stage antibodies specifically detecting proteins of the extracellular signal regulated kinase Erk 1/2 cell signalling pathway and transcription factors. Tissue sections were collected from four natural cases of OPA, four experimentally induced OPA tumours, four ENA tumours in sheep, four ENA tumours in goats, two normal sheep lungs and two lungs with chronic inflammation. Routine immunohistochemical procedures with phosphorylation stage-specific antibodies were carried out. Representative proteins of the Erk1/2 pathway (Raf-1, Mek1/2 and p44/42MAPK) were activated in natural cases of OPA and ENA in sheep and goats and also in experimentally induced OPA. Transcription factors 90Rsk and Elk-1 were activated in OPA and ENA tumours. However, c-Myc was activated only in OPA tumours. In contagious respiratory neoplasms of sheep and goats the Erk1/2 pathway appears to be important for the in-vivo generation of the transforming stimuli.

Jaagsiekte sheep retrovirus-specific immune responses induced by vaccination: A comparison of immunisation strategies

Jaagsiekte sheep retrovirus (JSRV) is the aetiological agent of ovine pulmonary adenocarcinoma (OPA). No JSRV-specific immunological responses have been detected in clinical cases of OPA or in experimentally infected lambs. The aim of the present study was to induce immune responses in sheep against JSRV proteins using several immunisation strategies. The vaccines were administered subcutaneously and intradermally, or intranasally, in adjuvant. Antibodies were measured by ELISA and immunoblotting, and T cell responses by lymphoproliferation assay. Antibodies specific for JSRV-capsid protein were induced by inoculation of recombinant proteins in adjuvant, and transient JSRV-specific T cell responses by intranasal inoculation with inactivated virus. These results will help in the design of a protective vaccine against JSRV infection and the development of OPA.

Molecular biology of jaagsiekte sheep retrovirus.

Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a contagious lung cancer of sheep. Until recently, research on JSRV/OPA was hampered by the lack of a tissue culture system for the propagation of the virus. Historically, pathological samples (lung fluid) collected from sheep affected by OPA were the only source of infectious JSRV. Thus studies on the JSRV/OPA system were conducted only where field isolates of OPA cases were readily available. In the past 10 years, the deduction of the JSRV sequence (York et al. 1991; York 1992), the isolation of an infectious and oncogenic JSRV molecular clone (JSRV21) (Palmarini et al. 1999a) and the establishment of a rapid method to produce infectious virus in vitro (Palmarini et al. 1999a) sparked many studies at the molecular level that strengthened past observations and revealed new properties of this unique virus. Here, we will review the data accumulated so far on the molecular biology of JSRV using the infectious and oncogenic JSRV21 molecular clone as virus of reference.

Alveolar type II cells expressing jaagsiekte sheep retrovirus capsid protein and surfactant proteins are the predominant neoplastic cell type in ovine pulmonary adenocarcinoma.

Ovine pulmonary adenocarcinoma is caused by jaagsiekte sheep retrovirus. To gain insight into the histogenesis and viral pathogenesis of this neoplasm, the tumor cell phenotypes and differentiation state were correlated with the distribution of jaagsiekte sheep retrovirus capsid protein in neoplastic and normal cells of the lung in nine naturally occurring and 12 experimentally induced cases of ovine pulmonary adenocarcinoma. Overall, 82% of tumor cells had ultrastructural features consistent with alveolar type II cells, 7% of tumor cells had features of Clara cells, and 11% of tumor cells were insufficiently differentiated to classify. The proportion of the neoplastic cell phenotypes varied within tumors, and no tumor consisted of a morphologically uniform cell population. To further characterize the neoplastic cell population, sections of tumors were immunostained with antibodies to surfactant protein A, surfactant protein C, and Clara cell 10-kd protein. Overall, surfactant proteins A and C were expressed in 70% and 80% of tumor cells, respectively, whereas Clara cell 10-kd protein was expressed in 17% of tumor cells. Jaagsiekte sheep retrovirus capsid protein was detected in 71% of tumor cells and in macrophages (5/21 tumors examined) and in nonneoplastic alveolar and bronchiolar cells (6/14 tumors). Expression of this viral protein in neoplastic cells, classified morphologically and by immunophenotyping primarily as of the alveolar type II lineage, implies an important role for specific virus-cell interactions in the pathogenesis of ovine pulmonary adenocarcinoma.