CAJ | 학술논문

探索以犬腺病毒1型疫苗株(Cannaught Laboratory Limited.CLL)作为病毒重组疫苗和基因转移载体的可行性。方法构建带增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)报告基因的E3缺失重组病毒CLLEGFP。将CLLEGFP感染各种人源细胞，并以灌胃、腹腔注射、尾静脉注射和肌肉注射等不同途径接种昆明小鼠。多时间点取小鼠组织标本，冷冻干燥切片，观察EGFP的表达。4周后采集小鼠血清，以Western blot分析抗EGFP 抗体的产生。结果 CLLEGFP能够感染各种人源细胞并表达EGFP。在腹腔接种CLLEGFP 3 d的小鼠肝组织细胞中可见转导的EGFP。Western blot分析显示，以各种途径免疫接种重组病毒4周后的小鼠血清中均存在抗EGFP特异抗体。结论 CLL具有开发成为病毒重组疫苗和基因转移载体的潜力。
탐색이견선병독1형역묘주(Cannaught Laboratory Limited.CLL)작위병독중조역묘화기인전이재체적가행성。방법구건대증강형록색형광단백(enhanced green fluorescent protein,EGFP)보고기인적E3결실중조병독CLLEGFP。장CLLEGFP감염각충인원세포，병이관위、복강주사、미정맥주사화기육주사등불동도경접충곤명소서。다시간점취소서조직표본，냉동간조절편，관찰EGFP적표체。4주후채집소서혈청，이Western blot분석항EGFP 항체적산생。결과 CLLEGFP능구감염각충인원세포병표체EGFP。재복강접충CLLEGFP 3 d적소서간조직세포중가견전도적EGFP。Western blot분석현시，이각충도경면역접충중조병독4주후적소서혈청중균존재항EGFP특이항체。결론 CLL구유개발성위병독중조역묘화기인전이재체적잠력。
Objective To evaluate canine adenovirus type 1 vaccine strain (Cannaught Laboratory Limited,CLL) as recombinant vaccine and gene transfer vector. Methods Recombinant virus CLLEGFP which contains enhanced green fluorescent protein(EGFP) reporter gene was constructed. CLLEGFP was used to infect various human derived cell lines (293, Hela, CO, SW, Hep-2 and CAM) by inoculating intraperitoneally(IP), intravenously(IV)and intramuscularly (IM)to Kunming mice other than oral administration. Various tissue samples of the mice were collected at multitime point for observing EGFP green fluorescence. Anti-EGFP antibodies were detected by Western blot analysis in the sera after 4 weeks. Results CLLEGFP can infect various human derived cell lines and express EGFP. EGFP green fluorescence were observed in liver tissue cells after IP transducing 3 days. All immune inoculation ways above could induce Kunming mice producing anti-EGFP antibodies which were identified by Western blot analysis. Conclusions These resluts indicate that CLL possess powerful potential as recombinant vaccine and gene transfer vector.