Glycoengineering of the hepatitis C virus E2 glycoprotein improves biochemical properties and enhances immunogenicity.

IF 6.5 1区 医学 Q1 IMMUNOLOGY
Liudmila Kulakova, Kiki H Li, Austin W T Chiang, Michael P Schwoerer, Saori Suzuki, Sanne Schoffelen, Khadija H Elkholy, Kinlin L Chao, Salman Shahid, Bhoj Kumar, Nathan B Murray, Stephanie Archer-Hartmann, Parastoo Azadi, Bjørn G Voldborg, Alexander Marin, Roy A Mariuzza, Alexander K Andrianov, Alexander Ploss, Nathan E Lewis, Eric A Toth, Thomas R Fuerst
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Abstract

An effective vaccine against hepatitis C virus (HCV) must elicit the production of broadly neutralizing antibodies (bnAbs) reproducibly against the E1E2 glycoprotein complex. Little is known about how glycan content affects this process. Ideally, glycans would maximize epitope exposure without compromising antigen stability or exposing new epitopes. However, typical recombinant vaccines contain considerable heterogeneity in glycan content, which can affect the antibody response and neutralization potency. Here we employed glycoengineered Chinese hamster ovary (geCHO) cell lines that impart nearly homogeneous glycosylation as a means to test how specific glycan features influence antigenicity and immunogenicity for the secreted HCV E2 ectodomain (sE2). Specific geCHO antigens exhibited a modest but reproducible increase in affinity for some mAbs relative to CHO- and HEK293-produced sE2. Surprisingly, one geCHO sE2 antigen failed to bind the CD81 receptor, indicating the potential for significant glycan effects on biochemical properties. We immunized mice with the four antigens and found the total antibody response to be the same for all groups. However, sera from one geCHO group exhibited a 7-fold improvement in neutralization against the homologous HCV pseudovirus (HCVpp) and had the most mice whose sera exhibited neutralization activity against genotypes 1b, 2a, 2b, and 3. Further analysis identified beneficial and deleterious glycan features, and the glycan that correlated the most with decreased potency was relatively small. However, size was not the sole determinant of glycan-driven effects on the antibody response. In summary, glycan content impacts biochemical properties of antigens to varying degrees and such effects can influence immune response quality and uniformity.

丙型肝炎病毒E2糖蛋白的糖工程改善生化特性,增强免疫原性。
一种有效的丙型肝炎病毒(HCV)疫苗必须引起可复制的广泛中和抗体(bnAbs)的产生,以对抗E1E2糖蛋白复合物。关于聚糖含量如何影响这一过程,我们所知甚少。理想情况下,聚糖可以在不影响抗原稳定性或暴露新表位的情况下最大化表位暴露。然而,典型的重组疫苗在多糖含量上存在相当大的异质性,这可能会影响抗体反应和中和效力。在这里,我们使用糖工程中国仓鼠卵巢细胞系(geCHO)进行几乎均匀的糖基化,作为测试特定聚糖特征如何影响分泌的HCV E2外结构域(sE2)的抗原性和免疫原性的手段。特异性的geCHO抗原显示出相对于CHO-和hek293产生的sE2,对一些单克隆抗体的亲和力适度但可重复地增加。令人惊讶的是,一个geCHO sE2抗原未能结合CD81受体,这表明可能存在显著的聚糖对生化特性的影响。我们用这四种抗原免疫小鼠,发现所有组的总抗体反应是相同的。然而,一个geCHO组的血清对同源HCV假病毒(HCVpp)的中和作用提高了7倍,并且对基因型1b、2a、2b和3表现出中和活性的小鼠最多。进一步分析确定了有益和有害的聚糖特征,与效力降低相关最大的聚糖相对较小。然而,大小并不是甘聚糖驱动抗体反应的唯一决定因素。综上所述,多糖含量不同程度地影响抗原的生化特性,从而影响免疫反应的质量和均匀性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
NPJ Vaccines
NPJ Vaccines Immunology and Microbiology-Immunology
CiteScore
11.90
自引率
4.30%
发文量
146
审稿时长
11 weeks
期刊介绍: Online-only and open access, npj Vaccines is dedicated to highlighting the most important scientific advances in vaccine research and development.
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