Nogueira R D, King W F, Gunda G, Culshaw S, Taubman M A, Mattos-Graner R O, Smith D J
Piracicaba Dental School, University of Piracicaba-UNICAMP, Piracicaba, SP, Brazil.
Infect Immun. 2008 Aug;76(8):3606-13. doi: 10.1128/IAI.00214-08. Epub 2008 May 12.
The interplay between mucosal immune responses to natural exposure to mutans streptococci and the incorporation and accumulation of these cariogenic microorganisms in oral biofilms is unclear. An initial approach to explore this question would be to assess the native secretory immunity emerging as a consequence of Streptococcus mutans infection. To this end, we analyzed salivary immunoglobulin A (IgA) antibody to mutans streptococcal glucosyltransferase (Gtf) and glucan binding protein B (GbpB) and to domains associated with enzyme function and major histocompatibility complex (MHC) class II binding in two experiments. Salivas were collected from approximately 45-day-old Sprague-Dawley rats, which were then infected with S. mutans SJ32. Infection was verified and allowed to continue for 2 to 2.5 months. Salivas were again collected following the infection period. Pre- and postinfection salivas were then analyzed for IgA antibody activity using peptide- or protein-coated microsphere Luminex technology. S. mutans infection induced significant levels of salivary IgA antibody to Gtf (P < 0.002) and GbpB (P < 0.001) in both experiments, although the levels were usually far lower than the levels achieved when mucosal immunization is used. Significantly (P < 0.035 to P < 0.001) elevated levels of postinfection salivary IgA antibody to 6/10 Gtf peptides associated with either enzyme function or MHC binding were detected. The postinfection levels of antibody to two GbpB peptides in the N-terminal region of the six GbpB peptides assayed were also elevated (P < 0.031 and P < 0.001). Interestingly, the patterns of the rodent response to GbpB peptides were similar to the patterns seen in salivas from young children during their initial exposure to S. mutans. Thus, the presence of a detectable postinfection salivary IgA response to mutans streptococcal virulence-associated components, coupled with the correspondence between rat and human mucosal immune responsiveness to naturally presented Gtf and GbpB epitopes, suggests that the rat may be a useful model for defining mucosal responses that could be expected in humans. Under controlled infection conditions, such a model could prove to be helpful for unraveling relationships between the host response and oral biofilm development.
口腔黏膜对变形链球菌自然暴露的免疫反应与这些致龋微生物在口腔生物膜中的定植和积累之间的相互作用尚不清楚。探索这个问题的初步方法是评估因变形链球菌感染而产生的天然分泌免疫。为此,我们在两个实验中分析了唾液中针对变形链球菌葡糖基转移酶(Gtf)、葡聚糖结合蛋白B(GbpB)以及与酶功能和主要组织相容性复合体(MHC)II类结合相关结构域的免疫球蛋白A(IgA)抗体。从约45日龄的斯普拉格-道利大鼠中收集唾液,然后用变形链球菌SJ32感染。感染得到验证并持续2至2.5个月。感染期结束后再次收集唾液。然后使用肽或蛋白包被的微球Luminex技术分析感染前后唾液中的IgA抗体活性。在两个实验中,变形链球菌感染均诱导唾液中针对Gtf(P < 0.002)和GbpB(P < 0.001)产生显著水平的IgA抗体,尽管这些水平通常远低于黏膜免疫时所达到的水平。检测到感染后唾液中针对6/10种与酶功能或MHC结合相关的Gtf肽段的IgA抗体水平显著升高(P < 0.035至P < 0.001)。在所检测的6种GbpB肽段N端区域的两种GbpB肽段的感染后抗体水平也有所升高(P < 0.031和P < 0.001)。有趣的是,啮齿动物对GbpB肽段的反应模式与幼儿初次接触变形链球菌时唾液中的模式相似。因此,感染后唾液中可检测到针对变形链球菌毒力相关成分的IgA反应,以及大鼠和人类黏膜对天然呈现的Gtf和GbpB表位的免疫反应性之间的对应关系,表明大鼠可能是一个有用的模型,用于定义人类预期的黏膜反应。在受控感染条件下,这样的模型可能有助于阐明宿主反应与口腔生物膜发育之间的关系。