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）和分泌型伊加（sIgA）对口腔健康的影响， 生态学产生了不一致的结果。 很难定义 这些抗微生物蛋白在体内的作用的措施，它也 很难对可能影响这些因素的因素进行控制， 行动 拟议的研究将在临床上解决这些问题 Lz、Lf、Spx和sIgA与血链球菌结合的研究 口腔内的单层细胞。 研究将设计有对照， 抗菌蛋白口服清除率的变化，相互作用 抗菌蛋白之间，细菌对抗菌剂的亲和力 蛋白质，以及抗菌蛋白在不同口腔组织中的定位 网站. 研究将按照这些具体目标确定的阶段进行： (1)联合收割机将吞咽活动的电子记录与敏感的 酶联免疫测定法（ELISA），以开发用于定量的方案 Lz、Lf、Spx和sIgA在单次吞咽周期中分泌。 （二） 联合收割机将该方案与多元统计方法相结合， 200或40例Lz、Lf、Spx水平较高或较低的受试者人群， 和sIgA被召回用于体内研究。 (3)提供结果 通过开发ELISA来测量Lz、Lf、Spx， 和与S结合的sIgA。在牛牙釉质芯片上形成的血单层。 （四） 用芯片ELISA法筛选一组沙门氏菌。两株血吸虫分离株 其在Lz、Lf、Spx和sIgA的结合方面有很大不同。 (5)贯彻 体内研究通过将芯片与S.前部和后部的血单层 在上下颚的后牙。 Lz、Lf、Spx和 将在受试者之间比较对高和低亲和力菌株的sIgA 以前发现，在一个特定的细胞中分泌高或低水平的这些蛋白质。 吞咽周期 有待检验的假设是， Lz、Lf、Spx和sIgA决定这些蛋白质与口腔细菌的结合 在体内，当由于口腔部位、细菌菌株 唾液清除率和抗菌蛋白之间的相互作用， 控制。 结果可能会澄清先前的临床解释 问题研究 开发的方法也将在未来有广泛的应用 研究抗菌蛋白在体内的作用。

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.