目的:人乳头瘤病毒(HPV)感染在全球范围内非常普遍。虽然存在预防性HPV疫苗,需要治疗性疫苗来治疗现有的HPV病变和恶性肿瘤.这项研究评估了三种基于重组蛋白的治疗性疫苗候选物的免疫刺激和抗肿瘤作用,肿瘤细胞裂解物(TCL),和工程化的外泌体(Exo)在小鼠模型中具有热休克蛋白27(Hsp27)-E7融合构建体。
方法:首先,重组Hsp27-E7蛋白在大肠杆菌表达系统中产生。然后,使用慢病毒系统产生基于肿瘤细胞裂解物和基于工程外泌体的含有绿色荧光蛋白(GFP)和Hsp27-E7的疫苗构建体。最后,在预防和治疗实验中研究了它们的免疫和抗肿瘤作用。
结果:我们的数据显示重组Hsp27-E7蛋白,TCL-Hsp27-E7和Exo-Hsp27-E7方案可以诱导最高水平的IFN-γ,TNF-α和颗粒酶B,分别。在预防和治疗实验中,三种疫苗策略(存活率:75%)的无瘤小鼠百分比相同。一般来说,TCL-Hsp27-E7,Exo-Hsp27-E7和重组Hsp27-E7蛋白方案诱导了对Th1和CTL活性的有效免疫应答,以及随后在小鼠模型中的抗肿瘤作用。
结论:关于更高的粒酶B分泌,更低的肿瘤生长和更多的安全性,Exo-Hsp27-E7方案可被认为是最有希望的HPV疫苗接种策略.
OBJECTIVE: Human papillomavirus (HPV) infections are highly prevalent globally. While preventive HPV vaccines exist, therapeutic vaccines are needed to treat existing HPV lesions and malignancies. This study evaluated the immunostimulatory and anti-tumor effects of three therapeutic vaccine candidates based on the recombinant protein, tumor cell lysate (TCL), and engineered exosome (Exo) harboring the heat shock protein 27 (Hsp27)-E7 fusion construct in mouse model.
METHODS: At first, the recombinant Hsp27-E7 protein was generated in E. coli expression system. Then, tumor cell lysates-based and engineered exosomes-based vaccine constructs harboring green fluorescent protein (GFP) and Hsp27-E7 were produced using lentiviral system. Finally, their immunological and antitumor effects were investigated in both prophylactic and therapeutic experiments.
RESULTS: Our data showed that the recombinant Hsp27-E7 protein, TCL-Hsp27-E7 and Exo-Hsp27-E7 regimens can induce the highest level of IFN-γ, TNF-α and Granzyme B, respectively. The percentage of tumor-free mice was identical for three vaccine strategies (survival rate: 75 %) in both prophylactic and therapeutic experiments. Generally, the TCL-Hsp27-E7, Exo-Hsp27-E7 and recombinant Hsp27-E7 protein regimens induced effective immune responses toward Th1 and CTL activity, and subsequently antitumor effects in mouse model.
CONCLUSIONS: Regarding to higher Granzyme B secretion, lower tumor growth and more safety, the Exo-Hsp27-E7 regimen can be considered as the most promising HPV vaccination strategy.