Kreider J W, Cladel N M, Patrick S D, Welsh P A, DiAngelo S L, Bower J M, Christensen N D
Department of Pathology, Milton S. Hershey Medical Center, Pennsylvania State University, Hershey 17033, USA.
J Virol Methods. 1995 Oct;55(2):233-44. doi: 10.1016/0166-0934(95)00062-y.
Plasmids containing cottontail rabbit papillomavirus (CRPV) DNA can induce papillomas in vivo, but efficiency has been low. The aim of the present investigation was to explore some of the technical variables involved in inoculation of rabbits with recombinant CRPV DNA in attempts to improve both yield and consistency of papilloma induction. It was found that induction of epidermal hyperplasia, with either a mixture of turpentine and acetone or phorbol esters, produced a marked increase in papilloma yield. An additional powerful factor was the use of very vigorous, cutaneous scarification, sufficient to penetrate the papillary dermis and produce bleeding. When used in combination, papilloma yields were consistent and often reached 90-100% of inoculated sites. A number of other variables which did not consistently affect papilloma yield were tested. These included bleb and puncture injections, plasmid dose, vector type, occlusive dressings, lipofection reagent, carrier DNA, and different methods for plasmid DNA extraction and purification. It is concluded that the most important variables in improving papilloma yields were prior induction of epidermal hyperplasia and vigorous cutaneous scarification.
含有棉尾兔乳头瘤病毒（CRPV）DNA的质粒可在体内诱导乳头瘤，但效率一直较低。本研究的目的是探索在给兔子接种重组CRPV DNA过程中涉及的一些技术变量，以提高乳头瘤诱导的产量和一致性。研究发现，用松节油和丙酮的混合物或佛波酯诱导表皮增生，可使乳头瘤产量显著增加。另一个重要因素是使用非常剧烈的皮肤划痕法，足以穿透乳头真皮并导致出血。当联合使用时，乳头瘤产量稳定，且通常达到接种部位的90-100%。还测试了一些其他对乳头瘤产量没有持续影响的变量。这些变量包括水疱注射和穿刺注射、质粒剂量、载体类型、封闭敷料、脂质体转染试剂、载体DNA以及质粒DNA提取和纯化的不同方法。得出的结论是，提高乳头瘤产量的最重要变量是预先诱导表皮增生和剧烈的皮肤划痕法。
