Reglinski Mark, Ercoli Giuseppe, Plumptre Charlie, Kay Emily, Petersen Fernanda C, Paton James C, Wren Brendan W, Brown Jeremy S
1Department of Respiratory Medicine, Centre for Inflammation and Tissue Repair, University College London, London, UK.
2Department of Pathogen Molecular Biology, London School of Hygiene and Tropical Medicine, London, UK.
NPJ Vaccines. 2018 Oct 31;3:53. doi: 10.1038/s41541-018-0090-4. eCollection 2018.
The pneumococcal conjugate vaccine (PCV) strongly protects against vaccine serotypes, but the rapid expansion of non-vaccine serotype disease and the vaccine's high expense has reduced its overall impact. We have developed Protein Glycan Coupling Technology (PGCT) as a flexible methodology for making low-cost polysaccharide/protein glycoconjugates recombinantly in . We have used PGCT to make a recombinant PCV containing serotype 4 capsular polysaccharide linked to the proteins NanA, PiuA, and Sp0148. The introduction of the UDP-glucose 4-epimerase gene GalE () into improved the yield of the resulting glycoprotein. PGCT glycoconjugate vaccination generated strong antibody responses in mice to both the capsule and the carrier protein antigens, with the PiuA/capsule glycoconjugate inducing similar anti-capsular antibody responses as the commercial PCV Prevnar-13. Antibody responses to PGCT glycoconjugates opsonised and expressing the serotype 4 capsule and promoted neutrophil phagocytosis of to a similar level as antisera generated by vaccination with Prevnar-13. Vaccination with the PGCT glycoconjugates protected mice against meningitis and septicaemia with the same efficacy as vaccination with Prevnar-13. In addition, vaccination with the protein antigen components from PGCT glycoconjugates alone provided partial protection against septicaemia and colonisation. These data demonstrate that a vaccine made by PGCT is as effective as Prevnar-13, identifies PiuA as a carrier protein for glycoconjugate vaccines, and demonstrates that linking capsular antigen to protein antigens has additional protective benefits that could provide a degree of serotype-independent immunity.
肺炎球菌结合疫苗（PCV）能有效预防疫苗血清型感染，但非疫苗血清型疾病的迅速增加以及该疫苗的高昂成本降低了其整体效果。我们开发了蛋白聚糖偶联技术（PGCT），这是一种灵活的方法，可用于在[具体生物体系]中重组制备低成本的多糖/蛋白糖缀合物。我们利用PGCT制备了一种重组PCV，其包含与蛋白NanA、PiuA和Sp0148连接的4型荚膜多糖。将UDP - 葡萄糖4 - 差向异构酶基因GalE（[具体基因形式]）导入[具体生物体系]提高了所得糖蛋白的产量。PGCT糖缀合物疫苗接种在小鼠体内引发了针对荚膜和载体蛋白抗原的强烈抗体反应，其中PiuA/荚膜糖缀合物诱导的抗荚膜抗体反应与商业PCV沛儿13相似。对PGCT糖缀合物的抗体反应可调理表达4型荚膜的[具体细菌1]和[具体细菌2]，并促进中性粒细胞对[具体细菌1]的吞噬作用，达到与接种沛儿13产生的抗血清相似的水平。用PGCT糖缀合物疫苗接种可保护小鼠免受脑膜炎和败血症感染，其效力与接种沛儿13相同。此外，单独用PGCT糖缀合物中的蛋白抗原成分进行疫苗接种可提供对败血症和定植的部分保护。这些数据表明，由PGCT制备的疫苗与沛儿13一样有效，确定PiuA为糖缀合物疫苗的载体蛋白，并表明将荚膜抗原与[具体蛋白抗原]连接具有额外的保护益处，可提供一定程度的血清型非依赖性免疫。
