靶向PreS1的HBV mRNA疫苗的开发与免疫原性评价

Development and evaluation of PreS1-targeted HBV mRNA vaccine

目的通过靶向HBV PreS1抗原设计并制备具有强免疫原性的治疗性mRNA疫苗。方法基于中国患者常见的GenotypeC-Adr亚型HBV毒株的PreS1抗原序列,并添加T7 RNA聚合酶启动子、UTR序列和分泌肽,设计并通过不同平台优化mRNA疫苗序列。将mRNA包封在LNP中,通过肌肉注射免疫小鼠。通过ELISA检测血清中的抗体水平,ELISpot检测脾脏T细胞的抗原特异性应答验证mRNA疫苗的免疫原性。结果与空白对照组相比,mRNA疫苗组小鼠血清中产生大量PreS1特异性IgG抗体(P<0.05)。ELISpot结果显示,mRNA疫苗组脾脏T细胞对PreS1肽库刺激特异性产生IFNγ。结论靶向PreS1的治疗性HBV mRNA疫苗具有良好的免疫原性,在小鼠体内诱导特异性B细胞反应(体液免疫)和T细胞反应(细胞免疫)。

Objective To design and develop a therapeutic mRNA vaccine encoding HBV PreS1 antigen with strong immunogenicity.Methods Based on the PreS1 antigen sequence of GenotypeC-Adr subtype HBV strains that commonly infect Chinese patients,mRNA vaccine sequences was designed and optimized using different platforms with the addition of T7 RNA polymerase promoter,UTR sequence,and signal peptide.The mRNA was then efficiently encapsulated with an ionizable lipid-based LNP,and mice were immunized via intramuscular injection.Subsequently,antibody levels in serum were detected using ELISA,and antigen-specific responses of splenic T cells were validated by ELISpot to assess the immunogenicity of the mRNA vaccine.Results Compared to the control group,mice in the mRNA vaccine groups showed significantly increased levels of PreS1-specific IgG antibodies in serum(P<0.05).The ELISpot assay demonstrated that splenic T cells from the mRNA vaccine groups produced IFNγin response to stimulation with PreS1 peptide pools.Conclusion The therapeutic HBV mRNA vaccine encoding PreS1 antigen exhibited robust immunogenicity,inducing specific B cell(humoral immunity)and T cell responses(cellular immunity)in mice.