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HPV16 E6、E7多表位DNA疫苗的构建及免疫效果评估

崔湘杰 陶玉芬 朱兰芳 姚宇峰 史荔

崔湘杰, 陶玉芬, 朱兰芳, 姚宇峰, 史荔. HPV16 E6、E7多表位DNA疫苗的构建及免疫效果评估[J]. 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20240503
引用本文: 崔湘杰, 陶玉芬, 朱兰芳, 姚宇峰, 史荔. HPV16 E6、E7多表位DNA疫苗的构建及免疫效果评估[J]. 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20240503
Xiangjie CUI, Yufen TAO, Lanfang ZHU, Yufeng YAO, Li SHI. Evaluations of Immunogenicity and Efficacy of A Novel HPV16 E6 and E7 Multi-epitope DNA Vaccine[J]. Journal of Kunming Medical University. doi: 10.12259/j.issn.2095-610X.S20240503
Citation: Xiangjie CUI, Yufen TAO, Lanfang ZHU, Yufeng YAO, Li SHI. Evaluations of Immunogenicity and Efficacy of A Novel HPV16 E6 and E7 Multi-epitope DNA Vaccine[J]. Journal of Kunming Medical University. doi: 10.12259/j.issn.2095-610X.S20240503

HPV16 E6、E7多表位DNA疫苗的构建及免疫效果评估

doi: 10.12259/j.issn.2095-610X.S20240503
基金项目: 中国医学科学院医学与健康科技创新工程基金资助项目(2021-I2M-1-004);云南省重大科技专项基金资助项目(202002AA100009)
详细信息
    作者简介:

    崔湘杰(1998 ~ ),女,辽宁铁岭人,在读硕士研究生,主要从事感染性疾病及肿瘤的免疫遗传学研究工作

    通讯作者:

    史荔,E-mail:shili.imb@gmail.com

  • 中图分类号: R737.33

Evaluations of Immunogenicity and Efficacy of A Novel HPV16 E6 and E7 Multi-epitope DNA Vaccine

  • 摘要:   目的  构建和评价HPV16 E6、E7多表位DNA疫苗诱导的特异性CTL细胞应答及其对肿瘤生长的干预作用,从而揭示其作为候选HPV治疗性疫苗的潜能。  方法  首先通过IEDB网站中的MHC I Processing Predictions和MHC I Binding Predictions方法,分别预测人类HLA-A*02:01、HLA-A*11:01、HLA-A*24:02和C57BL/6小鼠H-2b的限制性CTL表位,然后根据评分以及ELISPOT实验筛选出二者共同呈递的CTL表位,并将其构建成多表位DNA疫苗(pVAX1-10P)。从预防性和治疗性二个方面研究pVAX1-10P对小鼠移植TC-1异位癌的免疫干预作用,流式细胞术检测特异性CTL应答。  结果  获得10条可被人与鼠MHC分子共呈递的CTL表位,ELISPOT结果表明这10条CTL表位均能诱导小鼠淋巴细胞产生特异性免疫应答;由此构建的多表位DNA疫苗pVAX1-10P无论在预防性实验还是治疗性实验中,均能诱导特异性的细胞免疫并抑制肿瘤的生长。  结论  构建的HPV16 E6、E7多表位DNA疫苗pVAX1-10P能够诱导特异性CTL应答,显著抑制肿瘤生长,有望作为候选HPV治疗性DNA疫苗。
  • 图  1  小鼠免疫程序

    A:疫苗的预防性抗肿瘤评价中的小鼠免疫程序;B:疫苗的治疗性抗肿瘤评价中的小鼠免疫程序。

    Figure  1.  Immunisation program in mice

    图  2  ELISPOT检测结果

    A:实验组及对照组代表性酶联免疫斑点法图;B:ELISPOT检测IFN-γ斑点数统计图,条形图以$ \bar x \pm s $表示。

    Figure  2.  ELISPOT results

    图  3  质粒构建图

    Figure  3.  Plasmid construction diagram

    图  4  多表位DNA疫苗诱导的预防性抗肿瘤免疫

    A:肿瘤解剖图;B:流式细胞术检测的CD8+T IFN-γ占比;与对照组比较,*P < 0.05。

    Figure  4.  Prophylactic anti-tumor immunity induced by multi-epitope vaccine

    图  5  多表位DNA疫苗诱导的治疗性抗肿瘤免疫

    A:肿瘤解剖图;B:流式细胞术检测的CD8+T IFN-γ占比;与对照组比较,*P < 0.05,nsP ˃ 0.05。

    Figure  5.  Therapeutic anti-tumor immunity induced by multi-epitope vaccine

    表  1  预测得到的人鼠共呈递的CTL表位

    Table  1.   Predicted human and mouse co-presented CTL epitopes

    蛋白 肽名称 起始位点 结束位点 序列 总分(鼠) 总分(人) 等级排名(鼠) 等级排名(人)
    E6 E6p1 38 45 VYCKQQLL −0.81 −1.39 1.4 0.45
    E6p2 49 57 VYDFAFRDL −1.39 −1.23 1.3 0.28
    E6p3 75 83 KFYSKISEY −0.72 −0.74或−0.75 1.5 1.6
    E6p4 124 132 RHLDKKQRF −1.3 −1.06 1.4 0.46
    E6p5 81 90 SEYRHYCYSL −0.29 1.7
    82 90 EYRHYCYSL −0.68 0.57
    E6p6 86 95 YCYSLYGTTL −0.52 −0.37 1.6 0.41
    E7 E7p1 49 57 RAHYNIVTF 0.83或−0.59 −0.3 0.01或0.6 0.54
    E7p2 7 15 TLHEYMLDL −1.08 −0.28 1.3 0.21
    E7p3 50 57 AHYNIVTF −0.04或−1.26 −1.12 0.23或1.2 1.2
    E7p4 77 87 RTLEDLLMGTL −1.37 −1.36 1.3 1.2
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  • 收稿日期:  2024-01-16
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