Key Laboratory for Oral Biomedical Engineering of Ministry of Education, School & Hospital of Stomatology, Wuhan University, China.
Acta Pharmacol Sin. 2013 Feb;34(2):239-46. doi: 10.1038/aps.2012.145. Epub 2012 Dec 31.
To investigate the effects of anti-caries DNA vaccine-induced salivary secretory immunoglobulin A (S-IgA) antibodies on Streptococcus mutans (S. mutans) adherence and biofilms formation in vitro.
Adult female Wistar rats were intranasally immunized with the anti-caries DNA vaccine pGJA-P/VAX. Their saliva samples were collected at different times after the immunization, and S-IgA antibody level in the saliva and its inhibition on S. mutans adherence were examined. The effects of S-IgA in the saliva with the strongest inhibitory effects were examined at 3 different stages, ie acquired pellicles, biofilm formation and production of mature biofilms. The number of viable bacteria and depth of the biofilm at 16 h in each stage were determined using counting colony forming units and using a confocal laser scanning microscopy (CLSM). The participation of S-IgA in acquired pellicles and its aggregation with S. mutans were also observed under CLSM.
The S-IgA titer in saliva reached its peak and exhibited the strongest inhibition on S. mutans adhesion at 10 weeks after the immunization. The colonies and depth of the biofilm in the saliva-pretreated group were 41.79% and 41.02%, respectively, less than the control group. The colonies and depth of the biofilm in the co-culture group were 27.4% and 22.81% less than the control group. The assembly of S. mutans and S-IgA was observed under CLSM after co-cultivation. In the mature-stage biofilm, no differences were observed between the different groups.
These results demonstrate that the anti-caries DNA vaccine induces the production of specific S-IgA antibodies that may prevent dental caries by inhibiting the initial adherence of S. mutans onto tooth surfaces, thereby reducing the accumulation of S. mutans on the acquired pellicles.
研究抗龋 DNA 疫苗诱导的唾液分泌型免疫球蛋白 A(S-IgA)抗体对变形链球菌(S. mutans)黏附及体外生物膜形成的影响。
成年雌性 Wistar 大鼠经鼻腔免疫抗龋 DNA 疫苗 pGJA-P/VAX。免疫后不同时间收集唾液样本,检测唾液中 S-IgA 抗体水平及其对 S. mutans 黏附的抑制作用。在获得性菌膜、生物膜形成和成熟生物膜产生的 3 个不同阶段,检测具有最强抑制作用的唾液 S-IgA 的作用。采用菌落形成单位计数法和共聚焦激光扫描显微镜(CLSM)检测各阶段 16 h 活菌数和生物膜深度。在 CLSM 下观察 S-IgA 在获得性菌膜中的参与及其与 S. mutans 的聚集。
免疫后 10 周,唾液中 S-IgA 滴度达到峰值,对 S. mutans 黏附的抑制作用最强。唾液预处理组的菌落数和生物膜深度分别为对照组的 58.67%和 58.12%,共培养组的菌落数和生物膜深度分别为对照组的 72.40%和 71.63%。CLSM 下观察到共培养后 S. mutans 和 S-IgA 的聚集。在成熟生物膜阶段,各实验组间无差异。
这些结果表明,抗龋 DNA 疫苗诱导产生特异性 S-IgA 抗体,可能通过抑制 S. mutans 初始黏附于牙面,从而减少获得性菌膜上 S. mutans 的积累,来预防龋病。