Palmarini M, Cousens C, Dalziel R G, Bai J, Stedman K, DeMartini J C, Sharp J M
Moredun Research Institute, Edinburgh, United Kingdom.
J Virol. 1996 Mar;70(3):1618-23. doi: 10.1128/JVI.70.3.1618-1623.1996.
Sheep pulmonary adenomatosis ([SPA] ovine pulmonary carcinoma) is a transmissible lung cancer of sheep that has been associated etiologically with a type D- and B-related retrovirus (jaagsiekte retrovirus (JSRV]). To date it has been impossible to cultivate JSRV in vitro and therefore to demonstrate the etiology of SPA by a classical approach. In addition, the presence of 15 to 20 copies of endogenous JSRV-related sequences (enJSRV) has hampered studies at the molecular level. The aim of this study was to investigate whether the expression of exogenous JSRV was specifically associated with neoplasia in SPA-affected animals. Initially, we found that enJSRVs were transcribed in a wide variety of normal sheep tissues. Then, by sequencing part of the gag gene of enJSRV we established a ScaI restriction site in gag as a molecular marker for the exogenous form of JSRV. Restriction enzyme digestion of PCR products obtained from the amplification of cDNA from a total of 65 tissues collected from SPA-affected and unaffected control sheep revealed that the exogenous form of JSRV was exclusively and consistently present in tumor tissues and lung secretions of the affected animals. In addition, exogenous JSRV provirus was detected only in DNA from SPA tumors and not from nontumor tissues of the same animals. This study has demonstrated clearly that the exogenous form of JSRV is specifically associated with SPA tumors.
绵羊肺腺瘤病([SPA]绵羊肺癌)是一种绵羊的传染性肺癌,在病因上与一种D型和B型相关逆转录病毒(绵羊肺腺瘤逆转录病毒[JSRV])有关。迄今为止,尚无法在体外培养JSRV,因此无法通过经典方法证明SPA的病因。此外,15至20个内源性JSRV相关序列(enJSRV)的存在阻碍了分子水平的研究。本研究的目的是调查外源性JSRV的表达是否与受SPA影响动物的肿瘤形成特异性相关。最初,我们发现enJSRV在多种正常绵羊组织中转录。然后,通过对enJSRV的gag基因部分进行测序,我们在gag中建立了一个ScaI限制性酶切位点作为外源性JSRV的分子标记。对从受SPA影响和未受影响的对照绵羊收集的总共65个组织的cDNA扩增得到的PCR产物进行限制性酶切分析,结果显示外源性JSRV仅在受影响动物的肿瘤组织和肺分泌物中持续存在。此外,仅在SPA肿瘤的DNA中检测到外源性JSRV前病毒,而在同一动物的非肿瘤组织中未检测到。这项研究清楚地表明,外源性JSRV与SPA肿瘤特异性相关。