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杆状病毒表达系统表达嵌合兔出血症病毒样颗粒,作为针对经典兔出血症病毒(GI.1)和兔出血症病毒2型(GI.2)的二价疫苗候选物。

The Baculovirus Expression System Expresses Chimeric RHDV VLPs as Bivalent Vaccine Candidates for Classic RHDV (GI.1) and RHDV2 (GI.2).

作者信息

Wang Yan, Fan Yiyang, Bi Ruixiang, Zhao Yapeng, Gao Wanning, Zhang Derong, Bai Jialin

机构信息

Engineering Research Center for Key Technology and Industrialization of Cell-Based Vaccine, Ministry of Education, Northwest Minzu University, Lanzhou 730030, China.

Key Laboratory of Bioengineering and Biotechnology of the National Ethnic Affairs Commission, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China.

出版信息

Vaccines (Basel). 2025 Jun 27;13(7):695. doi: 10.3390/vaccines13070695.

Abstract

BACKGROUND

Rabbit hemorrhagic disease (RHD) is an acute, hemorrhagic and highly lethal infectious disease caused by rabbit hemorrhagic disease virus (RHDV), which causes huge economic losses to the rabbit breeding industry. Moreover, there is limited cross-protection between the two different serotypes of classic RHDV (GI.1) and RHDV2 (GI.2). The shortcomings of traditional inactivated vaccines have led to the development of novel subunit vaccines that can protect against both strains, and the VP60 capsid protein is the ideal antigenic protein. This study focused on developing a bivalent RHDV vaccine that can prevent infection with both GI.1 and GI.2 strains.

METHODOLOGY

Baculovirus vectors containing classic RHDV and RHDV2 VP60 were co-transfected with linearized baculovirus into sf9 cells and transferred to baculovirus via homologous recombination of the VP60 gene. Infected sf9 cells were lysed, and after purification via Ni-NTA chromatography, VLPs were observed using transmission electron microscopy (TEM). In order to evaluate the immunogenicity of the chimeric RHDV VLP vaccine in rabbits, the RHDV VP60-specific antibody, IL-4, IFN-γ and neutralizing antibody titers were analyzed in serum using ELISA and HI.

RESULTS

The recombinant baculovirus system successfully expressed chimeric RHDV VLPs with a diameter of 32-40 nm. After immunization, it could produce specific antibodies, IL-4 and IFN-γ. Following the second immunization, neutralizing antibodies, determined using hemagglutination inhibition (HI) assays, were elicited.

CONCLUSIONS

These data show that the chimeric RHDV VLP bivalent vaccine for immunized New Zealand rabbits can induce humoral immunity and cellular immunity in vivo, and the immunization effect of the high-dose group is similar to that of the current commercial vaccine.

摘要

背景

兔出血症(RHD)是由兔出血症病毒(RHDV)引起的一种急性、出血性且致死率高的传染病,给养兔业造成巨大经济损失。此外,经典RHDV的两种不同血清型(GI.1)和RHDV2(GI.2)之间的交叉保护作用有限。传统灭活疫苗的缺点促使了能预防两种毒株感染的新型亚单位疫苗的研发,而VP60衣壳蛋白是理想的抗原蛋白。本研究聚焦于开发一种能预防GI.1和GI.2毒株感染的二价RHDV疫苗。

方法

将含有经典RHDV和RHDV2 VP60的杆状病毒载体与线性化杆状病毒共转染至sf9细胞,并通过VP60基因的同源重组转移至杆状病毒中。裂解感染的sf9细胞,经镍-氮三乙酸(Ni-NTA)层析纯化后,用透射电子显微镜(TEM)观察病毒样颗粒(VLPs)。为评估嵌合RHDV VLP疫苗在兔体内的免疫原性,采用酶联免疫吸附测定(ELISA)和血凝抑制试验(HI)分析血清中的RHDV VP60特异性抗体、白细胞介素-4(IL-4)、干扰素-γ(IFN-γ)及中和抗体效价。

结果

重组杆状病毒系统成功表达了直径为32 - 40 nm的嵌合RHDV VLPs。免疫后,其能产生特异性抗体、IL-4和IFN-γ。第二次免疫后,通过血凝抑制试验检测到诱导产生了中和抗体。

结论

这些数据表明,用于免疫新西兰兔的嵌合RHDV VLP二价疫苗可在体内诱导体液免疫和细胞免疫,且高剂量组的免疫效果与目前的商业疫苗相似。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a886/12299046/c2ed505a5cc8/vaccines-13-00695-g001.jpg

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