Induction of mucosal tolerance in Peyer‘s patch—deficient, ligated small bowel loops
Thomas A. Kraus, … , Peter Boros, Lloyd Mayer
Thomas A. Kraus, … , Peter Boros, Lloyd Mayer
Published August 1, 2005
Citation Information: J Clin Invest. 2005;115(8):2234-2243. https://doi.org/10.1172/JCI19102.
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Research Article Immunology

Induction of mucosal tolerance in Peyer‘s patch—deficient, ligated small bowel loops

Abstract

To explore the requirement for M cells and the Peyer’s patch (PP) in induction of oral tolerance and address the potential in vivo role of intestinal epithelial cells as nonprofessional APCs, we have attempted to induce tolerance in mice with ligated small bowel loops without M cells and Peyer’s patches. A 2-centimeter section of vascularized small bowel was spliced away from the gut without disruption of the mesenteric attachments. We introduced OVA directly into the lumen of the loop prior to footpad immunization. By excising segments of bowel that contain PPs in some mice and segments without patches in others, we could study the necessity of the M cell and the underlying patch versus epithelial cells in induction of mucosal tolerance. We show that OVA-specific T cell proliferation and serum antibody responses are reduced in mice that have previously been given OVA both in PP-containing loops and in loops without patches. Furthermore, both high- and low-dose tolerance could be induced in the absence of PPs. Low-dose tolerance is associated with bystander suppression and requires IL-10, which indicates active suppression and the induction of regulatory cells. These data suggest that there is a critical role for components of the mucosal immune system other than PPs in antigen sampling and induction of oral tolerance.

Authors

Thomas A. Kraus, Jens Brimnes, Christine Muong, Jian-Hua Liu, Thomas M. Moran, Kelly A. Tappenden, Peter Boros, Lloyd Mayer

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Induction of low-dose oral tolerance in normal mice. (A) Antigen-specifi...
Induction of low-dose oral tolerance in normal mice. (A) Antigen-specific T cell proliferation in nonfed and OVA-fed mice. The tolerance protocol was performed as described in Methods. *P < 0.035. (B) Serum anti-OVA antibody levels in nonfed and OVA-fed mice. Serum from mice that were (open circles) or were not (filled circles) fed OVA prior to OVA immunization was diluted (1.25 × 10–5 to 5 × 10–5) and analyzed for the presence of anti-OVA IgG by ELISA. Serum from a nonimmunized animal (normal mouse serum [NMS]; open squares) was used as a negative control. The titers of OVA-specific IgG were significantly lower in mice fed OVA compared with nonfed mice. #P < 0.007. (C) Cytokine profile of cells from OVA-fed or nonfed mice. Cells from the PLN of OVA-fed (black bars) and nonfed (white bars) OVA-immunized mice were cultured with 100 μg/ml OVA for 24–96 hours, and cytokine secretion was measured by ELISA. The results indicate that the OVA-fed mice had increased secretion of IL-4 (72 hours) and IL-10 (72 hours) and a decreased secretion of IFN-γ (24 hours) and IL-2 (24 hours) compared with nonfed mice.