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Tumor Necrosis Factor- in Serum of Patients with Inflammatory Bowel Disease as Measured by a Highly Sensitive Immuno-PCR

^aAddress correspondence to this author at: Department of Clinical Laboratory Medicine, Sapporo Medical University, School of Medicine, South-1, West-16, Chuo-Ku, Sapporo 060-8543, Japan. Fax 81-11-622-7502; e-mail watanabn{at}sapmed.ac.jp.

Background: The significance of serum concentrations of tumor necrosis factor- (TNF-) in the pathogenesis of inflammatory bowel disease (IBD) is uncertain. We measured TNF- in serum from IBD patients by immuno-PCR to analyze the relationship between TNF- and pathophysiologic state in IBD.

Methods: Serum samples were collected from 54 healthy blood donors, 29 patients with ulcerative colitis (UC; 46 samples), and 7 patients with Crohn disease (CD; 8 samples). DNA label was generated by PCR amplification using biotinylated primer and was bound with streptavidin to biotinylated third antibody. TNF- sandwiched by antibodies was detected by PCR amplification of the DNA label.

Results: TNF- could be measured in all samples. The median serum concentration in IBD patients overall was 390-fold higher than in healthy donors (median increase, 380-fold for UC, 640-fold for CD). The median serum TNF- concentration was 1.7-fold higher in the active stage of UC than in the inactive stage (P <0.05), and this difference could be detected in individual patients.

Conclusions: Sensitive measurement of serum TNF- could provide an important pathophysiologic marker for the presence and activity of IBD.

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