中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2012年
3期
276-280
,共5页
任皎%赵莉%田厚文%高见%冯靖%庞正%吴小兵%谭文杰%阮力
任皎%趙莉%田厚文%高見%馮靖%龐正%吳小兵%譚文傑%阮力
임교%조리%전후문%고견%풍정%방정%오소병%담문걸%원력
人乳头瘤病毒16型%宫颈癌%免疫治疗%腺病毒载体
人乳頭瘤病毒16型%宮頸癌%免疫治療%腺病毒載體
인유두류병독16형%궁경암%면역치료%선병독재체
Human papillomavirus type 16%Cervical cancer%Immunotherapy%Adenovirus vector
目的 构建用于宫颈癌治疗的HPV16E7E6重组腺病毒并评价其免疫效果.方法 根据哺乳细胞使用密码子的偏嗜性设计HPV16E7E6融合蛋白表达优势密码子基因序列并合成.将合成的E7E6基因插入腺病毒重组穿棱质粒pCD316后,与Ad5腺病毒骨架质粒共转染293细胞,重组病毒经单斑纯化并进行目的基因插入及表达的鉴定.扩增纯化重组病毒后免疫小鼠,评价其免疫活性.结果 PCR试验证明E7E6密码子优化基因成功插入Ad5病毒；Western blot检测结果表明重组腺病毒中E7E6优化基因能高效表达,该重组腺病毒免疫C57小鼠后,可诱发强特异性T细胞免疫应答,在小鼠体内能完全抑制5×104 TC-1移植瘤细胞的生长.结论 重组HPV16E7E6腺病毒可作为治疗HPV慢性感染或宫颈癌的候选疫苗.
目的 構建用于宮頸癌治療的HPV16E7E6重組腺病毒併評價其免疫效果.方法 根據哺乳細胞使用密碼子的偏嗜性設計HPV16E7E6融閤蛋白錶達優勢密碼子基因序列併閤成.將閤成的E7E6基因插入腺病毒重組穿稜質粒pCD316後,與Ad5腺病毒骨架質粒共轉染293細胞,重組病毒經單斑純化併進行目的基因插入及錶達的鑒定.擴增純化重組病毒後免疫小鼠,評價其免疫活性.結果 PCR試驗證明E7E6密碼子優化基因成功插入Ad5病毒；Western blot檢測結果錶明重組腺病毒中E7E6優化基因能高效錶達,該重組腺病毒免疫C57小鼠後,可誘髮彊特異性T細胞免疫應答,在小鼠體內能完全抑製5×104 TC-1移植瘤細胞的生長.結論 重組HPV16E7E6腺病毒可作為治療HPV慢性感染或宮頸癌的候選疫苗.
목적 구건용우궁경암치료적HPV16E7E6중조선병독병평개기면역효과.방법 근거포유세포사용밀마자적편기성설계HPV16E7E6융합단백표체우세밀마자기인서렬병합성.장합성적E7E6기인삽입선병독중조천릉질립pCD316후,여Ad5선병독골가질립공전염293세포,중조병독경단반순화병진행목적기인삽입급표체적감정.확증순화중조병독후면역소서,평개기면역활성.결과 PCR시험증명E7E6밀마자우화기인성공삽입Ad5병독；Western blot검측결과표명중조선병독중E7E6우화기인능고효표체,해중조선병독면역C57소서후,가유발강특이성T세포면역응답,재소서체내능완전억제5×104 TC-1이식류세포적생장.결론 중조HPV16E7E6선병독가작위치료HPV만성감염혹궁경암적후선역묘.
Objective To construct one recombinant adenovirus AdE7E6 expressing HPV16 E6 and E7 fusion protein as candidate for HPV16 therapeutic vaccine.Methods The codon-optimized E6 and E7 gene,were fused to create one open reading frame,then inserted into adenovirus vector pCD316.A strain of recombinant adenovirus was constructed through homologous recombinant in 293 cells,and identified by PCR and Western blot.Finally,it was employed to study it's immunogenicity and the activity of the tumor growth regression.Results The PCR result showed that E6E7 fusion gene had been integrated in recombinant Ad5 DNA.Western blot test confirmed that the E6E7 fusion protein was highly expressed in 293 cells infected with Ad5E7E6 recombinant adenovirus.The recombinant adenovirus elicited significant E7 specific CD8+ T lymphocyte response in vaccinated mice.These responses could completely prevent the TG-1 tumor cell bearing mice treated with AdE7E6 from developing into tumor.Conclusion These results suggested that rAd5E7E6 could be a potent vaccine candidate for the treatment of HPV16-associated tumors and their precancerous transformations.
