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Simultaneous assessment of complement components C3, C4, and B and their cleavage products in human gingival fluid
Author(s) -
Niekrash Christine E.,
Patters Mark R.
Publication year - 1985
Publication title -
journal of periodontal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.31
H-Index - 83
eISSN - 1600-0765
pISSN - 0022-3484
DOI - 10.1111/j.1600-0765.1985.tb00433.x
Subject(s) - periodontitis , agarose , radial immunodiffusion , complement system , chemistry , medicine , pathology , dentistry , chromatography , antibody , immunology
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.