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Adequacy of mucosal biopsies for evaluation of intestinal cytokine-specific mRNA

Comparative study of RT-PCR in biopsies and isolated cells from normal and inflamed intestine

  • Intestinal Disorders, Inflammatory Bowel Disease, Immunology, and Microbiology
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Abstract

Endoscopic biopsies are being increasingly utilized to investigate cytokine profiles in normal and diseased intestine. To evaluate the adequacy of mucosal biopsies as a source of cytokine-specific mRNA, we measured their content of interleukin-1β (IL-1β) and interleukin-2 (IL-2) mRNA by reverse transcription-polymerase chain reaction and compared it to that of autologous lamina propria mononuclear cells in control and inflammatory bowel disease-involved specimens. High-quality total RNA was recovered more consistently from cell isolates than from biopsies. Interleukin-1β mRNA was reliably detected in both cell and biopsy samples, whereas IL-2 mRNA was measurable in all lamina propria cells but not always in biopsies. Compared to controls, levels of IL-1β were significantly elevated in Crohn's disease and ulcerative colitis cells and biopsies, and a weak but significant correlation existed between values derived from the two sources. In contrast, only ulcerative colitis cell isolates but not biopsies contained significantly reduced concentrations of IL-2 mRNA compared to those of control and Crohn's disease samples, and no correlation was found between cell and biopsy contents. Widely different levels of cytokine-specific mRNA were present in closely adjacent biopsies, particularly for IL-2. These results suggest that mucosal biopsies are suitable to assess some but not all cytokine-specific mRNA due to variation in RNA recovery, intrinsic level of cytokine gene expression, and substantial variability of cytokine transcripts.

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Authors and Affiliations

  1. Division of Gastroenterology, Case Western Reserve University School of Medicine, 10900 Euclid Avenue, 44106-4952, Cleveland, Ohio

    Kouhei Fukushima MD, PhD, Gail West BS &  Claudio Fiocchi MD

Authors
  1. Kouhei Fukushima MD, PhD
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  2. Gail West BS
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  3. Claudio Fiocchi MD
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This work was supported by a grant (DK 30399) from the National Institutes of Health (NIDDK).

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Fukushima, K., West, G. & Fiocchi, C. Adequacy of mucosal biopsies for evaluation of intestinal cytokine-specific mRNA. Digest Dis Sci 40, 1498–1505 (1995). https://doi.org/10.1007/BF02285198

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  • DOI: https://doi.org/10.1007/BF02285198

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