IN VIVO STUDIES OF SALIVA ANTIMICROBIAL PROTEINS

试图将唾液中溶菌酶(LZ)、乳铁蛋白(Lf)、 唾液过氧化物酶(Spx)和分泌性IgA(SIgA)与口腔健康和 生态学产生了不一致的结果。很难给它下定义 这些抗菌蛋白在体内的作用措施，它还 很难为可能影响这些因素的因素设计控制措施 行为。拟议的研究将在临床上解决这些问题 LZ、Lf、Spx、SIgA与血链球菌结合的研究 放在嘴里的单层。研究将设计为具有对照的 抗菌蛋白的口服清除、相互作用引起的变异 抗菌蛋白之间、细菌对抗菌剂的亲和力 蛋白质，以及不同口腔中抗菌蛋白的定位 网站。研究将分阶段进行，具体目标如下： (1)将吞咽活动的电子记录与敏感相结合 酶联免疫分析(ELISA法)开发一种定量检测 LZ、Lf、Spx和SIgA在单次吞咽周期中分泌。(2) 将该方案与多元统计方法相结合，以筛选出 LZ、Lf、Spx、LZ、Lf、Spx、 和SIgA将被召回进行体内研究。(3)提供结果 通过开发ELISA来测量LZ、Lf、Spx、 SIgA与牛牙釉质芯片上形成的血链球菌单层结合。(4) 用芯片ELISA法筛选两株血链球菌分离物 它们与LZ、Lf、Spx和SIgA的结合差异很大。(5)在 将含有血链球菌单层的芯片放置在前部和前部的活体研究 上颌和下颚的后牙。LZ、Lf、Spx和 高亲和力菌株和低亲和力菌株的SIgA将在受试者之间进行比较 先前发现，在体内分泌高水平或低水平的这些蛋白质 吞咽周期。需要检验的假设是唾液中的 Lz、Lf、Spx和SIgA决定这些蛋白质与口腔细菌的结合 在体内，当变异可归因于口腔部位、细菌菌株、 唾液清除，以及抗菌蛋白之间的相互作用是 控制住了。结果可能会澄清对先前临床的解释 学习。所开发的方法在未来也将有广泛的应用 体内抗菌蛋白作用的研究。

Attempts to relate saliva levels of lysozyme (Lz), lactoferrin (Lf), salivary peroxidase (Spx), and secretory IgA (sIgA) to oral health and ecology have yielded inconsistent results. It has been difficult to define measures of the action of these antimicrobial proteins in vivo, and it also has been difficult to devise controls for factors which may influence those actions. Proposed research will address those questions in clinical studies of Lz, Lf, Spx, and sIgA binding to Streptococcus sanguis monolayers placed in the mouth. Studies will be designed with controls for variation due to oral clearance of antimicrobial proteins, interaction between antimicrobial proteins, bacterial affinity for antimicrobial proteins, and localization of antimicrobial proteins in different oral sites. Research will proceed in stages defined by these specific aims: (1) Combine electronic recording of swallowing activity with sensitive enzyme-linked immunoassay (ELISA) to develop a protocol for quantitation of Lz, Lf, Spx, and sIgA secreted during single swallowing cycles. (2) Combine that protocol with multivariate statistical methods to screen a population of 200 or 40 subjects with high or low levels of Lz, Lf, Spx, and sIgA to be recalled for in vivo studies. (3) Provide an outcome variable for in vivo studies by developing ELISAs to measure Lz, Lf, Spx, and sIgA bound to S. sanguis monolayers formed on bovine enamel chips. (4) Use chip ELISAs to screen a panel of S. sanguis isolates for two strains which differ greatly in binding of Lz, Lf, Spx, and sIgA. (5) Carry out in vivo studies by placing chips with S. sanguis monolayers on anterior and posterior teeth in upper and lower jaws. The binding of Lz, Lf, Spx, and sIgA to high and low affinity strains will be compared between subjects previously found to secrete high or low levels of those proteins in a swallowing cycle. The hypothesis is to be tested is that saliva levels of Lz, Lf, Spx, and sIgA determine binding of those proteins to oral bacteria in vivo when variation attributable to oral sites, bacterial strains, saliva clearance, and interaction between antimicrobial proteins is controlled. Results may clarify the interpretation of previous clinical studies. Approaches developed also will have broad applications for future studies of antimicrobial protein action in vivo.