Partial characterization of an interleukin-1-like factor in human gingival crevicular fluid from patients with chronic inflammatory periodontal disease.
A significant level of interleukin-1-(IL-1)-like activity was detected in gingival crevicular fluid obtained from sites in patients with chronic inflammatory periodontal disease, confirming a previous report of IL-1-like activity detected in human gingival crevicular fluid from patients with chronic inflammatory periodontitis (J. A. Charon, T. A. Luger, S. E. Mergenhagen, and J. J. Oppenheim, Infect. Immun. 38:1190-1195, 1982). In the present study, we sought to investigate whether this IL-1-like activity belonged to IL-1 alpha or IL-1 beta and to characterize some of the biochemical properties of this factor. Polyclonal antibodies against recombinant human IL-1 alpha or IL-1 beta (rIL-1 alpha or rIL-1 beta) have been used for serological comparison of the IL-1-like factor. IL-1-like activity was completely neutralized by anti-human rIL-1 alpha antiserum, but not by anti-human rIL-1 beta antiserum. On gel filtration with a high-pressure liquid chromatographic Superose 12 column, IL-1-like activity was separated into two peaks, one with a molecular weight of about 43,000 and the other with a molecular weight of less than 17,000. The majority of the IL-1-like factor with a low molecular weight in human gingival crevicular fluid migrated at a molecular weight of about 17,000 under the reducing conditions of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The specificity of the IL-1-like factor was further confirmed by an immunochemical method (Western blotting [immunoblotting]) by using anti-human rIL-1 alpha monoclonal antibodies. On isoelectric chromatography with a high-pressure liquid chromatographic Mono P column, the pI of this IL-1-like factor was between pH 4.9 and 5.2. These results suggest that the IL-1-like factor in human gingival crevicular fluid from diseased sites in patients with chronic inflammatory periodontitis consists predominantly of IL-1 alpha.
Department of Periodontics and Endodontics, Faculty of Dentistry, Kyushu University, Fukuoka, Japan.
H
H
Kabashima
K
K
Maeda
Y
Y
Iwamoto
T
T
Hirofuji
M
M
Yoneda
K
K
Yamashita
M
M
Aono
D000328:Adult; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002850:Chromatography, Gel; D002908:Chronic Disease; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005883:Gingival Crevicular Fluid / Q000276:immunology*; D005891:Gingivitis / Q000276:immunology*; D006801:Humans; D007249:Inflammation; D007375:Interleukin-1 / Q000096:biosynthesis* / Q000302:isolation & purification; D007526:Isoelectric Point; D008297:Male; D008875:Middle Aged; D009500:Neutralization Tests; D010510:Periodontal Diseases / Q000276:immunology*; D011994:Recombinant Proteins
D000911:Antibodies, Monoclonal; D007375:Interleukin-1; D011994:Recombinant Proteins