皮下注射以CpG寡脱氧核苷酸为佐剂的人乳头瘤病毒多肽疫苗对小鼠原位宫颈癌的抑制作用研究  

The inhibitory effect of hppillomavirus peptide vaccine on cervical cancer in mice with CpG olidoxynucleotide as adjuvant

作　　者：王欢 王雪莲 车玉鑫 马壮 WANG Huan;WANG Xue-lian;CHE Yu-xin;MA Zhuang(Jinzhou Medical University Nursing College,Jinzhou,Liaoning 121001,China)

机构地区：[1]锦州医科大学护理学院,辽宁锦州121001

出　　处：《中国妇幼保健》2023年第2期329-333,共5页Maternal and Child Health Care of China

基　　金：辽宁省自然科学基金项目(2021-MS-322)。

摘　　要：目的研究皮下注射以CpG寡脱氧核苷酸为佐剂的人乳头瘤病毒(HPV)多肽疫苗对小鼠原位宫颈癌的抑制作用。方法采用6~8周C57BL/6小鼠,通过将TC-1细胞直接注入子宫颈附近的阴道黏膜下层来模拟原位宫颈癌的发生,建立小鼠原位宫颈癌模型,分析以CpG ODN作为佐剂的HPV E7多肽疫苗对小鼠肿瘤生长的抑制作用;应用流式细胞术研究以CpG ODN作为佐剂的HPV E7多肽疫苗对C57BL/6小鼠宫颈局部肿瘤微环境的影响;制备负载HPV 16 E7多肽和佐剂CpG ODN的甘露糖修饰脂质体疫苗,利用透射电镜观察甘露糖修饰的脂质体疫苗形态,利用激光粒度分析仪在水中测量脂质体疫苗的粒度分布和zeta电位,应用荷宫颈癌的小鼠,研究负载HPV 16 E7多肽和佐剂CpG ODN的甘露糖修饰脂质体疫苗对晚期宫颈癌的治疗作用;应用流式细胞术研究脂质体疫苗对小鼠产生特异性细胞免疫应答的影响;应用qRT-PCR研究肿瘤组织中的细胞因子、趋化因子及基质金属蛋白酶(MMP)等表达水平;应用免疫组织化学法研究肿瘤组织中的凋亡、侵袭、血管形成及胶原分布等对小鼠肿瘤微环境的影响。结果皮下注射CpG ODN为佐剂的HPV多肽治疗性疫苗,可显著增加肿瘤局部浸润的CD4+T细胞和CD8+T细胞,降低肿瘤局部的骨髓衍生抑制细胞(MDSCs)、肿瘤相关成纤维细胞(CAFs),调节一系列与肿瘤增殖、侵袭、转移及血管形成相关的细胞因子、趋化因子、MMPs等分子的表达水平。结论皮下注射CpG ODN为佐剂的HPV多肽治疗性疫苗可抑制已建立的原位宫颈转移瘤的生长。Objective To research the inhibitory effect of hppillomavirus(HPV)peptide vaccine on cervical cancer in mice with CpG olidoxynucleotide as adjuvant.Methods C57BL/6 mice for 6-8 weeks were used to simulate the occurrence of cervical cancer in situ by injecting TC-1 cells directly into the vaginal submucosa near the cervix.The mouse model of cervical cancer in situ was established to analyze the inhibitory effect of HPV E7 polypeptide vaccine with CpG ODN as adjuvant on tumor growth in mice.The effect of HPV E7 polypeptide vaccine with CpG ODN as adjuvant on the local tumor microenvironment of C57BL/6 mice was studied by flow cytometry.Mannose modified liposome vaccine loaded with HPV 16 E7 polypeptide and adjuvant CpG ODN was prepared.The morphology of mannose modified liposome vaccine was observed by transmission electron microscope.The particle size distribution and zeta potential of Liposome Vaccine were measured by laser particle size analyzer in water to study the therapeutic effect of mannose modified liposome vaccine loaded with HPV 16 E7 polypeptide and adjuvant CpG ODN on advanced cervical cancer.Flow cytometry was used to study the effect of Liposome Vaccine on specific cellular immune response in mice.The expression levels of cytokines,chemokines,and matrix metalloproteinase(MMP)in tumor tissues were studied by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).Immunohistochemical method was used to study the effects of apoptosis,invasion,angiogenesis,and collagen distribution on tumor microenvironment in mice.Results HPV peptide therapeutic vaccine with CpG ODN as adjuvant can significantly increase the locally immersed CD4+T cells and CD8+T cells of tumor,reduce bone marrow derived suppressor cells(MDSCs)and cancer associated fibroblasts(CAFs)in tumor,and regulate the expression levels of a series of cytokines,chemogenization factors,MMPs,and other molecules associated with tumor proliferation,invasion,metastasis,and vascular formation.Conclusion HPV peptide therapeutic vaccine with

关 键 词：CPG寡脱氧核苷酸 人乳头瘤病毒 多肽疫苗 宫颈癌 

分 类 号：R332[医药卫生—人体生理学]