Enzyme-linked immunosorbent assay for S100A9 in the stool of rats with dextran sulfate sodium-induced colitis.
Calprotectin, a heterodimer of S100A8 and S100A9, has been reported to be a useful biomarker in inflammatory bowel disease (IBD); however, the relationship between the fecal level of S100A9 and the extent of inflammation in IBD remains unclear. Our aim was to develop a new enzyme-linked immunosorbent assay (ELISA) for rat S100A9, and to investigate whether changes in fecal S100A9 levels reflect the inflammatory conditions in the intestinal tracts of rats with dextran sulfate sodium (DSS)-induced colitis. Anti-rat S100A9 monoclonal antibodies were raised in mice and used for the development of a novel ELISA for rat S100A9. The performance of our ELISA was assessed by dilution and recovery tests, and the detection range was 3.75-240ng/mL. The dilution test showed good linearity. The recovery of fecal S100A9 was 95.1% (mean), with a range of 86.1%-108.8%. Colitis was induced in rats by oral administration of 3% DSS/drinking water (DW) for 11days (D group), while DW alone was provided to rats of the control group (C group) during the same period. The extent of inflammation was evaluated with the disease activity index (DAI), and the concentration of fecal S100A9 was determined by ELISA. Both the DAI scores and the fecal S100A9 levels were significantly higher in the D group than in the C group. Microscopic observation revealed that S100A9 was dominantly produced in many immune cells of myeloid origin in rat rectal tissues. These results indicate that the novel ELISA may be applied to clinically evaluate IBD in rats with high sensitivity. In conclusion, our ELISA is useful in toxicological and pharmacological evaluations.
Research and Development Section, Diagnostics Department, Yamasa Corporation, 10-1 Araoi-cho 2-Chome, Choshi, Chiba 288-0056, Japan. Electronic address: sekiya@yamasa.com.
Shunsuke
S
Sekiya
Research and Development Section, Diagnostics Department, Yamasa Corporation, 10-1 Araoi-cho 2-Chome, Choshi, Chiba 288-0056, Japan.
Makoto
M
Murata
Research and Development Section, Diagnostics Department, Yamasa Corporation, 10-1 Araoi-cho 2-Chome, Choshi, Chiba 288-0056, Japan.
Satoshi
S
Arai
Research and Development Section, Diagnostics Department, Yamasa Corporation, 10-1 Araoi-cho 2-Chome, Choshi, Chiba 288-0056, Japan.
Hiroshi
H
Murayama
Research and Development Section, Diagnostics Department, Yamasa Corporation, 10-1 Araoi-cho 2-Chome, Choshi, Chiba 288-0056, Japan.
Atushi
A
Kawasaki
Research and Development Section, Diagnostics Department, Yamasa Corporation, 10-1 Araoi-cho 2-Chome, Choshi, Chiba 288-0056, Japan.
Noriyuki
N
Ashida
Department of Clinical Laboratory Science, Faculty of Health Care, Tenri Health Care University, 80-1 Bessho-cho, Tenri, Nara 632-0018, Japan; Human Health Sciences, Graduate School of Medicine, Kyoto University, 53 Shogoin-kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
Kohki
K
Okada
Department of Clinical Laboratory Science, Faculty of Health Care, Tenri Health Care University, 80-1 Bessho-cho, Tenri, Nara 632-0018, Japan.
Masaki
M
Ikemoto
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015415:Biomarkers / Q000378:metabolism; D040502:Calgranulin B / Q000276:immunology / Q000378:metabolism*; D003092:Colitis / Q000139:chemically induced / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D003106:Colon / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D016264:Dextran Sulfate; D004195:Disease Models, Animal; D018450:Disease Progression; D042241:Early Diagnosis; D004797:Enzyme-Linked Immunosorbent Assay; D005243:Feces / Q000737:chemistry*; D005260:Female; D008297:Male; D011237:Predictive Value of Tests; D011916:Rats, Inbred F344; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D040502:Calgranulin B; D016264:Dextran Sulfate
Dextran sulfate sodium-induced colitis; Enzyme-linked immunosorbent assay; Monoclonal antibody; S100A9; Stool