治疗型VP22△-mE6△/mE7重组蛋白疫苗的表达与免疫学初步分析

中国生物工程杂志

2009, Vol. 29

Issue (04): 6-11

研究报告

治疗型VP22△-mE6△/mE7重组蛋白疫苗的表达与免疫学初步分析

高丹丹,彭正华,杨旭,毕研伟,李智华,姬秋彦,李建芳,李健峰,徐维明

中国医学科学院昆明医学生物学研究所

Expression and Immune Effect Study of Therapeutic Recombinant Protein Vaccine VP22Δ-mE6Δ/mE7

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摘要： 制备16型人乳头瘤病毒mE6Δ/mE7蛋白与I型人单纯疱疹病毒VP22Δ蛋白的治疗型分子内佐剂融合蛋白疫苗，并检测其免疫原性和抗肿瘤相关生物活性。通过克隆HSV-1 VP22Δ及HPV-16 mE6Δ/mE7基因，构建pET28a-VP22Δ-mE6Δ/mE7原核表达载体。重组质粒在Rosetta（DE3）宿主菌中进行诱导表达，表达蛋白经分离、复性后，通过镍离子亲和层析进行纯化，纯化蛋白经SDS-PAGE、Western blot 鉴定，并免疫BalB/C及C57BL/6小鼠，检测其免疫原性和抗肿瘤活性。结果显示，VP22Δ-mE6Δ/mE7蛋白以包涵体形式表达，分子量约为34kDa，表达量约占菌体总蛋白的45%。该蛋白免疫小鼠后血清特异性IgG、特异性淋巴细胞增殖效果及对TC-1致瘤小鼠的肿瘤治疗效果均高于无佐剂单一重组蛋白疫苗。以上结果说明，所获得的重组融合蛋白具有较好的免疫原性和抗肿瘤活性，为治疗型HPV分子内佐剂疫苗的进一步研究奠定了基础。

关键词： HPV-16;重组蛋白疫苗;分子内佐剂;VP22△;mE6△/mE7

Abstract: In order to investigate the biological effects of the VP22Δ-mE6Δ/mE7 built in adjuvant fusion protein vaccine on the tumor associated with HPV-16 infection.HSV-1 VP22Δ and HPV-16 mE6Δ/mE7 genes were cloned,and the pET28a-VP22Δ-mE6Δ/mE7 recombinat prokaryotic expression vector was constructed.Vector was transformed into Rosetta(DE3)E.coli string and expressed under the induction of IPTG.The re-naturalized protein was then purified via Ni2+ affinity adsorbent column and identified by SDS-PAGE and Western blot.Purified protein was immunized BalB/C and C57BL/6 mice to evaluate the immunogenicity and anti-tumor activity.The expressed recombinant protein formed as inclusion body with a prediction MW about 34kDa and contained approximately 45% of total somatic protein.The VP22Δ-mE6Δ/mE7 immunization induced higher titer of specific IgG against HPV,higher level of lymphocyte proliferation and better effect on suppressing HPV16 positive TC-1 tumor growth than the mice immunized with mE6Δ/mE7 alone.The results showed that the recombined built-in adjuvant vaccine could induce specific cellular immune response in vitro and inhibit the TC-1 tumor proliferation in vivo,that would be a foundation for further studies and developments of inner adjuvant vaccines.

Key words: HPV-16;recombinant protein vaccine;built-in adjuvant;VP22△;mE6△/mE7

收稿日期: 2008-10-27 出版日期: 2009-04-27

ZTFLH:

Q786

基金资助:

云南省自然科学基金资助项目（2007C144M）;省部基金

通讯作者: 李健峰

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引用本文:

高丹丹,彭正华,杨旭,毕研伟,李智华,姬秋彦,李建芳,李健峰,徐维明. 治疗型VP22△-mE6△/mE7重组蛋白疫苗的表达与免疫学初步分析[J]. 中国生物工程杂志, 2009, 29(04): 6-11.

GAO Dan-Dan- Bang-Zheng-Hua- Yang-Xu- Bi-Xing-Wei- Li-Zhi-Hua- Ji-Qiu-Pan- Li-Jian-Fang- Li-Jian-Feng- Xu-Wei-Meng. Expression and Immune Effect Study of Therapeutic Recombinant Protein Vaccine VP22Δ-mE6Δ/mE7. China Biotechnology, 2009, 29(04): 6-11.

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https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2009/V29/I04/6

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