Alimentary TractIdentification of a novel bacterial sequence associated with Crohn's disease☆,☆☆
Section snippets
Patients
Lamina propria mononuclear cells were produced from a therapeutic colectomy specimen of a single Cedars-Sinai Medical Center patient with CD. Endoscopic biopsy specimens were obtained from 7 patients with CD at UCLA Medical Center. Paraffin blocks of ileal and colonic resection specimens were obtained from the surgical pathology archives of the UCLA and Cedar-Sinai Medical Centers. All available CD cases between 1993 and 1997 were evaluated, yielding 121 (61 colonic and 60 ileal), which
Cloning of gene segments associated with histologically involved CD mucosa
RDA was performed between lamina propria mononuclear cells derived from involved (lesional) and uninvolved (nonlesional) mucosa of a single CD colectomy specimen. After 3 cycles of RDA, 4 major DNA bands were obtained from the involved mucosa (designated I1 to I4), and 2 bands from the uninvolved mucosa (designated U1 and U2) (Figure 1A).
Discussion
This study describes a novel bacterial sequence, I2, isolated by subtractive cloning in lamina propria mononuclear cells from colonic mucosa histologically involved with CD. PCR analysis indicated that I2 DNA was commonly detectable in involved CD colonic mucosa, but was uncommon in control inflammatory and noninflammatory mucosa. Serum antibodies to an I2-encoded peptide were also observed to be disease associated.
Acknowledgements
The authors thank Christopher Denny for advice on RDA; Sydney Finegold for advice and efforts in bacterial culture; Gunther Harth, Diana Lester, and Marcus Horwitz for mycobacterial isolates and cultures; and Karin Reimann and Hallie Sandusky for technical assistance.
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Address requests for reprints to: Jonathan Braun, M.D., Ph.D., Department of Pathology and Laboratory Medicine, UCLA School of Medicine, CHS 13-222, Los Angeles, California 90095-1732. e-mail: [email protected]; fax: (310) 825-5674.
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Supported by National Institutes of Health grant DK46763, DK43026, the UCLA Clinical and Fundamental Immunology Training Grant (to C. L. S. and H. D., AI 07126-23), the Crohn's and Colitis Foundation of America, the Jonnson Comprehensive Cancer Center, and the Feintech Family Chair of Inflammatory Bowel Disease.