title: Jaagsiekte retrovirus establishes a disseminated infection of the lymphoid tissues of sheep affected by pulmonary adenomatosis.
abstract: Jaagsiekte retrovirus (JSRV) is an exogenous type D-related retrovirus specifically associated with a contagious lung cancer of sheep (sheep pulmonary adenomatosis; SPA). Recently, epithelial tumour cells in the lungs of SPA-affected sheep were identified as major sites of JSRV replication by immunological techniques and RT-PCR amplification of part of JSRV gag. JSRV was not detected outside the lungs and their draining lymph nodes. However, low levels of JSRV expression in non-respiratory tissues could have been masked by co-amplification of endogenous JSRV-related sequences, which were differentiated from JSRV by the lack of a Scal restriction site in the PCR product. To further investigate the pathogenesis of SPA, an exogenous virus-specific hemi-nested PCR was developed utilizing primers in the U3 region of JSRV LTR, where major differences between endogenous and exogenous sequences exist. This technique was shown to be > or = 10(5)-fold more sensitive than the previous gag PCR/ScaI digestion method. Using this new assay the tissue distribution of JSRV in sheep with natural and experimentally induced SPA was analysed. Proviral DNA and JSRV transcripts were found in all tumours and lung secretions of SPA-affected sheep (n = 22) and in several lymphoid tissues. The mediastinal lymph nodes draining the lungs were consistently demonstrated to be infected by JSRV (10/10). JSRV transcripts were also detected in spleen (7/9), thymus (2/4), bone marrow (4/8) and peripheral blood mononuclear cells (3/7). Proviral DNA was also detected in these tissues although in a much lower proportion of cases. JSRV was not detected in 27 samples from unaffected control animals (n = 15).
authors.affiliation: Moredun Research Institute, Edinburgh, UK.
authors.forename: M
authors.initials: M
authors.lastname: Palmarini
authors.forename: M J
authors.initials: MJ
authors.lastname: Holland
authors.forename: C
authors.initials: C
authors.lastname: Cousens
authors.forename: R G
authors.initials: RG
authors.lastname: Dalziel
authors.forename: J M
authors.initials: JM
authors.lastname: Sharp
mesh_terms: D000818:Animals; D001483:Base Sequence; D030121:Betaretrovirus / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000032:analysis; D008221:Lymphoid Tissue / Q000821:virology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000235:genetics; D011648:Pulmonary Adenomatosis, Ovine / Q000473:pathology / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis; D012091:Repetitive Sequences, Nucleic Acid; D012680:Sensitivity and Specificity; D012756:Sheep / Q000821:virology
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral