Simultaneous assessment of complement components C3, C4, and B and their cleavage products in human gingival fluid. I. Reliability of the method.
Simultaneous assessment of complement components C3, C4, and B and their cleavage products in human gingival fluid. I. Reliability of the method.
复制标题
同时评估人牙龈液中的补体成分 C3、C4 和 B 及其裂解产物。
DOI：
10.1111/j.1600-0765.1985.tb00433.x
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发表时间：
1985
影响因子：
3.5
通讯作者：
Patters,MR
中科院分区：
文献类型：
--
作者：
Niekrash,CE;Patters,MR
对补体系统在牙周病发病机制中的潜在作用的研究包括测量从牙周炎病变获得的牙龈液中的补体裂解。由于牙周病损可获得的牙龈液量有限，这些研究仅限于评估具有显著炎症的病变中的单一补体成分。本文介绍了一种改进的交叉免疫电泳法，并对该法的可靠性进行了评价。在0.2 μl的单个龈液样品中，使用多二维凝胶对收集在滤纸条上的牙龈液和应用于试纸条上的血清（作为对照）进行交叉免疫电泳，该凝胶由含有抗人B和抗人C3的2 cm宽琼脂糖平板组成，随后是含有抗人C4和抗人转铁蛋白（参考标记物）的另一个2 cm平板。干燥和染色后的凝胶，所得沉淀物进行测量和每一个组件的相对量和百分之C3的转化率determined.Using新鲜和活化的血清，这种技术被发现有一个在10%以内的变异。通过对26例牙周炎患者进行牙周治疗前重复取样，测定牙龈液中补体测量值的稳定性。在测量的任何组分中均未发现显著差异，表明该方法从临床样品中产生了可靠的结果。这些研究结果建立了这种方法作为一个敏感和可靠的手段，在牙龈液的单个样品中的补体裂解的评估。这种技术应该是有用的纵向测量的变化，在诱导或解决牙周病的补体。
Studies of the potential role of the complement system in the pathogenesis of periodontal disease have involved measurements of complement cleavage in gingival fluid obtained from lesions of periodontitis. Due to the limited amounts of gingival fluid available from periodontal lesions, these studies have been confined to assessment of single complement components in lesions with significant inflammation. This paper presents a description and an evaluation of reliability of an improved method of sampling and complement assessment by crossedimmunoelectrophoresis which permits measurement of C3, C4, B. and their conversion products in single gingival fluid samples as small as 0.2 μl. Gingival fluid collected on filter paper strips and serum applied to strips as controls were subjected to crossed‐immunoelectrophoresis using a multiple second‐dimension gel composed of a 2 cm wide slab of agarose with anti‐human B and anti‐human C3, followed by another 2 cm slab containing anti‐human C4 and antihuman transferrin (reference marker). Following drying and staining of the gels, the resultant precipitates were measured and the relative amount of each component and the percentage conversion of C3 determined.Using both fresh and activated serum, this technique was found to have a variability within 10%. Stability of complement measurements in gingival fluid was determined by duplicate sampling of 26 lesions of periodontitis prior to periodontal therapy. No significant differences were found in any of the components measured suggesting that the method produced reliable results from clinical samples. These findings establish this method as a sensitive and reliable means of assessment of complement cleavage in single samples of gingival fluid. This technique should be useful for longitudinal measurements of changes in complement during induction or resolution of periodontal disease.