Pathology and pathogenesis of ovine pulmonary adenocarcinoma.

Ovine pulmonary adenocarcinoma (OPA), also known as jaagsiekte, is a transmissible lung tumour of sheep caused by jaagsiekte sheep retrovirus (JSRV). JSRV induces neoplastic transformation of alveolar and bronchiolar secretory epithelial cells and the resulting tumours can grow to occupy a significant portion of the lung. Tumour growth is frequently accompanied by the overproduction of fluid in the lung, which further compromises normal respiration. The period between infection and the appearance of clinical signs may be several months or years and many JSRV-infected sheep do not exhibit clinical signs at all during their lifespan. This allows the spread of OPA into new flocks through contact with infected but apparently normal animals. OPA was first described in the early 19th century; however, it has still not been possible to devise effective methods for controlling its spread and it remains an important problem in most countries where sheep are farmed. This is due in part to the absence of an immunological response to JSRV in infected animals, which has hindered the development of serological diagnostic tests and vaccines. In addition to its veterinary importance, OPA is regarded as a potential large animal model for human lung adenocarcinoma and this has stimulated research into the pathogenesis of the ovine disease. This work has produced some significant results, including the finding that one of the JSRV structural proteins is directly involved in oncogenesis. The recent advances in understanding JSRV and the pathogenesis of OPA should lead to novel strategies for diagnosis and control of this disease and for its exploitation as a comparative model for human lung cancer.

A Moloney murine leukemia virus driven by the Jaagsiekte sheep retrovirus enhancers shows enhanced specificity for infectivity in lung epithelial cells.

Jaagsiekte sheep retrovirus (JSRV) is the etiologic agent of ovine pulmonary adenocarcinoma (OPA), a transmissible lung cancer in sheep. One of the unique features of this virus is that in infected animals, the only tissues that show expression of the virus are the tumor cells in the lung. We previously showed that the JSRV long terminal repeat (LTR) is preferentially active in murine lung epithelial cell lines (MLE-15 and mtCC1-2). To further explore the tissue specificity, we inserted the JSRV enhancer sequences from the U3 region of the LTR into a Moloney murine leukemia virus (M-MuLV) LTR lacking its own enhancer sequences, to give the chimeric LTR DeltaMo + JS. Transient transfection assays indicated that the DeltaMo + JS LTR is > 5-fold more active in lung epithelial cell lines than in non-lung lines, compared to the wild-type M-MuLV LTR. This was due to preferential activity of the JSRV enhancers in lung epithelial cells. Moreover, M-MuLV driven by the DeltaMo + JS LTR was > 3 logs more infectious in MLE-15 cells compared to non-lung cell lines. This chimeric virus may facilitate investigations of the tissue-specificity of JSRV.

Infection of lung epithelial cells and induction of pulmonary adenocarcinoma is not the most common outcome of naturally occurring JSRV infection during the commercial lifespan of sheep.

Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA). In this study, we followed over a 31-month period the natural transmission of JSRV in adult sheep and in their offspring. We established groups derived from flocks with either a high or low incidence of OPA and monitored virus transmission, clinical disease and macroscopic/microscopic lung lesions at necropsy. Results obtained show that (i) JSRV infection can occur perinatally or in the first few months of life in lambs and in adult sheep; (ii) only a minority of JSRV-infected animals develop clinical disease during their commercial lifespan; and (iii) JSRV is more readily detectable in peripheral blood leucocytes and lymphoid organs than in the lungs. These data support a model of opportunistic JSRV infection and tumorigenic conversion of type II pneumocytes/Clara cells in the lungs, while lymphoreticular cells serve as the principal virus reservoir.

An influx of macrophages is the predominant local immune response in ovine pulmonary adenocarcinoma.

Infection with a retrovirus, Jaagsiekte sheep retrovirus (JSRV), causes ovine pulmonary adenocarcinoma (OPA). The excess production of surfactant proteins by alveolar tumour cells results in increased production of pulmonary fluid, which is characteristically expelled through the nostrils of affected sheep. The immune response to JSRV and the tumour is poorly understood: no JSRV-specific circulating antibodies or T cells have been detected to date. The aim of the present study was to obtain phenotypic evidence for a local immune response in OPA lungs. Specific-pathogen free lambs were infected intratracheally with JSRV. When clinical signs of OPA were apparent, the lungs were removed at necropsy and immunohistochemistry (IHC) was performed on lung sections using a panel of mouse anti-sheep mAbs. No influx of dendritic cells, B cells, CD4, CD8 or gammadelta T cells was seen in the neoplastic nodules or in their periphery. MHC Class II-positive cells were found intratumourally, peritumourally and in the surrounding alveolar lumina. In the tumours, many of these cells were shown to be fibroblasts and the remainder were likely to be mature macrophages. In the alveolar lumen, the MHC Class II-positive cells were CD14-positive and expressed high levels of IFN-gamma. They appeared to be immature monocytes or macrophages which then differentiated to become CD14-negative as they reached the periphery of the tumours. A high level of MHC Class I expression was detected on a range of cells in the OPA lungs but the tumour nodules themselves contained no MHC Class I-positive cells. On the basis of these findings, it is proposed that the lack of an effective immune response in OPA could result from a mechanism of peripheral tolerance in which the activity of the invading macrophages is suppressed by the local environment, possibly as a consequence of the inhibitory properties of the surfactant proteins.

[New types of virus infections of domestic animals in the German Democratic Republic. 3. Seroepizootiologic and experimental studies of herpesvirus infections in sheep--the etiology of pulmonary adenomatosis]. / Für die DDR neuartige Virusinfektionen der Haustiere. 3. Mitteilung: Seroepizootiologische und experimentelle Untersuchungen zur Herpesvirusinfektion des Schafes--Ein Beitrag zur Atiologie der Lungenadenomatose.

Neutralisation tests for antibodies against ovine herpesvirus were applied to 848 sera which had been sampled from different sheep herds across the GDR. Between six and 20 percent of sheep in the herds tested exhibited neutralising antibodies, notwithstanding their pulmonary adenomatosis status. Incidence and titre distribution of antibodies against ovine herpesviruses in pulmonary adenomatosis herds were identical with those recorded from unsuspected herds. From among 21 sheep with pathomorphologically secured pulmonary adenomatosis, six animals exhibited antibody titres just as high as those recorded from responders of all herds examined. Lambs were obtained by hysterectomy and raised without mothers and were experimentally infected with Herpesvirus ovis. All of these animals responded to infection by clearly rising titres (between 1:2 and 1:32). Adenomatous pulmonary lesions were not recordable from any of them. One lamb, following experimental ovine herpesvirus infection, exhibition, exhibited subclinical interstitial pneumonia. Herpesvirus ovis has been widespread in sheep herds across the GDR. The authors' serological and experimental investigations do not support the assumption of an aetiological relationship between ovine herpesvirus infection and incidence of pulmonary adenomatosis.

Retrovirus-associated ovine pulmonary carcinoma (sheep pulmonary adenomatosis) and lymphoid interstitial pneumonia. I. Lesion development and age susceptibility.

To determine the lesion development of retrovirus-induced ovine pulmonary carcinoma (OPC), ten neonatal lambs were inoculated intratracheally with either 1) lung fluid preparations derived from a sheep with Type D retrovirus-associated OPC and concurrent ovine lentivirus (OvLV)-associated lymphoid interstitial pneumonia (LIP) (n = 8); or 2) lung fluid from a sheep with only OvLV-LIP (n = 2). Seven of eight neonates that received Type D retrovirus-associated OPC/OvLV-LIP lung fluid developed both OPC and LIP lesions between 9 and 32 weeks after inoculation. Mild OPC lesions consisted of foci of type II alveolar epithelial cells lining alveoli surrounded by minimal alveolar macrophage infiltrates. More severe OPC lesions consisted of multifocal aggregates of cuboidal to columnar neoplastic cells forming acini or masses associated with abundant alveolar macrophage infiltrates. Lesions of LIP consisted of peribronchiolar and perivascular lymphoid hyperplasia and heterogeneous interstitial leukocytic infiltrates. The two neonates that received OvLV-LIP lung fluid developed rapid and severe LIP, but not OPC lesions. Two lambs (inoculated as neonates with virus-free lung fluid) and three lambs (uninoculated contacts) served as controls and did not develop OPC. To investigate age susceptibility for development of OPC, 20 additional lambs within defined age groups (neonates, 2 weeks old, 5 weeks old, and 10 weeks old) received ultracentrifuged tumor homogenate. Neonatal to 5-week-old lambs inoculated with Type D retrovirus-associated OPC/OvLV-LIP tumor homogenate were equally likely to develop OPC, but lambs inoculated at 10 weeks of age were more refractory to tumor development.(ABSTRACT TRUNCATED AT 250 WORDS)

The possible involvement of immunosuppression caused by a lentivirus in the aetiology of jaagsiekte and pasteurellosis in sheep.

A South African isolate of ovine lentivirus was shown to cause a mild immunosuppression in sheep, reflected by a reduced delayed hypersensitivity reaction. This effect, measured in terms of skin swelling after intradermal inoculation with tuberculin, showed a positive linear relationship with the latency period before the appearance of jaagsiekte symptoms in animals co-infected with JSRV, as well as with the activity of monocytes. In a parallel study, increased susceptibility of lentivirus-infected sheep to infection with Pasteurella haemolytica was demonstrated. It is concluded that the lentivirus may play an enhancing role in both viral and bacterial infections of sheep by compromising the host's cellular immune response.

The etiology and pathogenesis of ovine pulmonary carcinoma (sheep pulmonary adenomatosis).

Ovine pulmonary carcinoma (OPC, sheep pulmonary adenomatosis, jaagsiekte) occurs naturally as a contagious bronchioloalveolar carcinoma of sheep in the Americas, Europe, Africa and Asia. The disease is endemic and economically important in Peru and apparently more common than previously suspected in the U.S.A. The tumor is a result of transformation of type II alveolar epithelial cells or non-ciliated bronchiolar cells of the lung. Clinically affected sheep develop dyspnea, tachypnea and often a watery nasal discharge that originates from tumor secretions. The course is progressive and death usually occurs within a few weeks. To study the viral etiology and pathogenesis of OPC in the U.S.A., the disease was experimentally transmitted to neonatal or young lambs with a success rate of 69%. Ovine lentivirus (OvLV), present in the inocula, was concurrently transmitted and induced lymphoid interstitial pneumonia in most animals. While morphological, immunological and other studies implicate a type D or type B retrovirus as the etiologic agent of OPC, this virus has not yet been cultured and the role of ovine lentivirus in the disease remains unknown.

[The so-called lung adenomatosis of sheep--an interesting pneumopathy from the comparative pathologic view]. / Die sogenannte Lungenadenomatose des Schafes--eine aus vergleichend pathologischer Sicht interessante Pneumopathie.

Contrary to conditions in human pathology, primary pulmonary neoplasms are rare in animals. The so-called pulmonary adenomatosis of sheep (PAS) is of greater importance among the epithelial lung tumors of domestic animals. PAS is a chronic, lethal pneumopathy caused by an oncogenic retrovirus. It is associated with the category of slow virus infections for its extremely long incubation period. PAS is characterized as a primary multicentric epithelial neoplasm which develops from proliferating alveolar pneumocytes II and non-ciliated bronchial epithelial cells. It has been recently classified as broncho-alveolar carcinoma on account of its morphological features and occasional metastasis into bronchial and mediastinal lymph nodes. PAS because of its similarity with human pulmonary carcinoma may be used as a model pneumopathy in comparative oncology.

Heterotransplantation of experimentally induced sheep lung adenomatosis into nude mice.

Male ICR nude mice inoculated subcutaneously with minced tissue obtained from pulmonary adenomatosis of sheep which was induced by RNA type-C retrovirus developed a transplant which contained cysts lined by a proliferative epithelium. These cysts showed several characteristics of the epithelial tumour cells of origin, such as high glycogen content and secretion of mucin. Microinvasion of epithelial cells from the adenomatous cysts into the stroma was observed. The stromal tissue demonstrated islets of cartilage. It was concluded that virus-induced pulmonary carcinoma of sheep can be transplanted and grown in nude mice.

Further evidence for a retrovirus as the aetiological agent of sheep pulmonary adenomatosis (jaagsiekte).

The infective agent of jaagsiekte was shown to be present in the fluid which accumulates in the respiratory tract of sheep during the terminal stages of the disease. The fluid also contained reverse transcriptase (RT) activity which showed a clear preference for a ribonucleic acid synthetic template over the corresponding deoxyribonucleic acid template and which utilised the RT specific template/primer poly (2'-0-methylcytidylate) oligodeoxyguanylate. This enzyme activity was associated with a particle which had typical retroviral buoyant densities in a range of gradient media.

Aetiology of jaagsiekte: transmission by means of subcellular fractions and evidence for the involvement of a retrovirus.

Jaagsiekte (ovine pulmonary adenomatosis) was transmitted to new-born lambs by inoculation of the microsomal fraction of a cytoplasmic extract of cultured tumour cells or tumour tissue. Various treatments of the biologically active fraction were carried out to differentiate between various classes of possible aetiological agents. The results obtained suggested the involvement of a membrane-associated RNA containing virus. Reverse transcriptase activity dependent on Mg++ was subsequently demonstrated in these extracts and in lung exudate, and was shown to be associated with particles banding at a density 1,175 in sucrose gradients. These characteristics, as well as the appearance of the particles in the electron microscope, are similar to those reported for Type B and Type D retroviruses. Serial transmissions of jaagsiekte over a number of years, using cytoplasmic extracts and purified virus, strongly suggest that this virus is the aetiologic agent of jaagsiekte.

Presence of Herpesvirus ovis DNA sequences in cellular DNA from sheep lungs affected with jaagsiekte (pulmonary adenomatosis).

To investigate further the possible involvement of Herpesvirus ovis in the aetiology of jaagsiekte, the kinetics of reassociation of viral DNA and DNA isolated from tumour tissue as well as from cell cultures derived from it were studied. Although DNA-DNA hybridization could be demonstrated in 2 cases of jaagsiekte, no correlation was found between the presence of Herpesvirus ovis genome sequences and the occurrence of the disease.

On the etiological role of Herpesvirus ovis in jaagsiekte.

Serologically related ovine herpesviruses have been isolated independently by various workers in different countries from adenomatous lung tissue of sheep or cell cultures derived from it. Although the disease can be transmitted with lung homogenates and with cell cultures, transmission attempts with virus alone failed. IUDR treatment of tumour-cell cultures and co-cultivation or fusion with cells permissive for virus replication induced antigens which react with some sera from tumour-bearing animals. These antigens give positive immunofluorescence reactions with sera both positive and negative in neutralization assays against the virus, however. Hybridization studies failed to demonstrate the presence of viral genomes in 15.4 tumour cells.

Characteristics of an ovine herpesvirus associated with pulmonary adenomatosis (jaagsiekte) in sheep.

The isolation of an ovine herpesvirus from a cell culture of an adenomatous sheep lung is reported, confirming previous observations of a possible association of a herpesvirus with this tumour. Some growth properties and morphological characteristics of the virus are described, as well as serological data supporting a possible relationship between tumour and virus. Attempts to produce jaagsiekte by intratracheal injection of virus into lambs were unsuccessful, suggesting that a second factor may be involved inthe oncogenic process possibly similar to that proposed for the well-known EBV-Burkitt's lymphoma system.