National Institute for Biological Standards and Control, Health Protection Agency, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK.
Vaccine. 2010 Apr 19;28(18):3211-8. doi: 10.1016/j.vaccine.2010.02.023. Epub 2010 Feb 25.
In this study, we evaluated the effect of the growth medium on the composition and immunogenicity of meningococcal outer membrane vesicle (OMV) vaccines after cultivation of the Norwegian serogroup B 44/76 vaccine strain in either Frantz' or modified Catlin-6 media (MC.6M). Differential proteomic analysis revealed that 97% of the OMV proteins maintained the same levels in the two preparations. However, a number of differentially expressed proteins, including TdfH, OpcA, OMP NMB0088, hypothetical NMB2134, lipoprotein NMB1126/1164 and NspA, increased significantly in OMVs produced from bacteria grown in the MC.6M. Together with increased lipopolysaccharide levels, the increased expression of these proteins was associated with significantly higher serum bactericidal titres in mice immunized with the MC.6M OMV vaccine. The high resolution two-dimensional separation of the OMVs on a large-format gel across a pH range of 3-11 resolved around 2000 protein spots from which 75 proteins were identified by mass spectrometry.
在这项研究中，我们评估了生长培养基对脑膜炎球菌外膜囊泡（OMV）疫苗组成和免疫原性的影响，方法是在 Frantz'或改良 Catlin-6 培养基（MC.6M）中培养挪威 B 群 44/76 疫苗株。差异蛋白质组学分析显示，两种制剂中 97%的 OMV 蛋白保持相同水平。然而，一些差异表达的蛋白质，包括 TdfH、OpcA、OMP NMB0088、假设的 NMB2134、脂蛋白 NMB1126/1164 和 NspA，在 MC.6M 中生长的细菌产生的 OMV 中显著增加。与脂多糖水平增加一起，这些蛋白质的高表达与用 MC.6M OMV 疫苗免疫的小鼠中的血清杀菌滴度显著增加有关。在 pH 范围为 3-11 的大格式凝胶上对 OMV 进行高分辨率二维分离，可从其中分离出约 2000 个蛋白质斑点，通过质谱鉴定出 75 种蛋白质。
