Gastroenterology

Gastroenterology

Volume 123, Issue 5, November 2002, Pages 1516-1526
Gastroenterology

Basic–Alimentary Tract
Expansion of CD8+ T cells with regulatory function after interaction with intestinal epithelial cells*,**

https://doi.org/10.1053/gast.2002.36588Get rights and content

Abstract

Background & Aims: Regulatory T cells play a role in the control of immune responses in the intestinal mucosa and their absence may predispose to inflammatory bowel disease (IBD). We have previously shown that T cells activated by intestinal epithelial cells (IECs) are suppressive in function. Our goal was to characterize the phenotype and function of T cells proliferating after interaction with IECs. Methods: Irradiated human IECs, isolated from normal resection specimens, were cultured with carboxy fluorescein succinimidyl ester (CFSE) labeled T cells. Flow cytometric analysis of T cells was performed at days 5–10. CD8+ T cells proliferating in culture with IECs were sorted and added to suppressive assays. Results: The precursor frequency of T cells proliferating in response to IECs ranged from 0.3%–0.9%. Several subpopulations were shown to proliferate (CD8+CD28−/CD8+CD28+/CD4+CD25+), but one population (CD8+CD28−CD101+CD103+) appeared to be dependent on contact with the CD8 ligand gp180. After sorting, culture in the presence of interleukin (IL)-7 and IL-15 allowed for the generation of cell lines. IEC-activated CD8+ T cells, but not nonactivated CD8+ T cells, were suppressive in function. Suppression belonged to the CD101+CD103+ subset of IEC-activated CD8+ T cells and appeared to require cell contact. CD8+ lamina propria T cells also showed suppressive function, suggesting the presence of CD8+ regulatory T cells in the mucosa. Conclusions: IECs are able to induce the proliferation of a small fraction of CD8+ peripheral T cells. The CD8+CD28− subset of IEC-activated CD8+ T cells, which express CD101 and CD103, interacts with IECs through gp180 and has regulatory function.

GASTROENTEROLOGY 2002;123:1516-1526

Section snippets

Preparation of human intestinal epithelial cells and lamina propria mononuclear cells

Surgical specimens from patients undergoing bowel resection for cancer (at least 10 cm away from tumor) at the Mount Sinai Medical Center were used as a source of IECs. IECs were isolated by a method described previously.18 Resected surgical specimens were washed extensively with phosphate-buffered saline (PBS). The mucosa was stripped off from the submucosa, minced into small pieces, and placed in 1 mmol/L dithiothreitol (Sigma Chemical Co., St. Louis, MO) for 10 minutes at room temperature to

Intestinal epithelial cells activate different subsets of T cells that represent less than 1% of the pool of peripheral blood T cells

To further study the interaction between IECs and lymphocytes, we used CFSE to assess the proliferation of T cells in IEC:T-cell cocultures. After 5 days of coculture, proliferating T cells (CD3+ CFSE low) represented 3.5%–35% of all T cells in more than 10 independent experiments (Figure 1).

. Quantitative analysis of T-cell proliferation in an IEC:T-cell coculture and calculation of the proportion of the initial population that has responded by dividing. T cells were labeled with CFSE before

Discussion

By using in vitro IEC:T-cell cocultures, we show that only a small fraction of peripheral T cells respond to IEC. IECs express several antigen-presenting molecules, such as classical class I and II molecules, nonclassical class Ib molecules (CD1d, human leukocyte antigen E, MICA/B, FcRn), and costimulatory molecules (gp180, B7h24). As expected, we found that IECs activated different subsets of T cells. Each of them may interact with a different combination of antigen-presenting and

References (35)

  • T Takahashi et al.

    Immunologic self-tolerance maintained by CD25(+)CD4(+) regulatory T cells constitutively expressing cytotoxic T lymphocyte-associated antigen 4

    J Exp Med

    (2000)
  • AM Thornton et al.

    CD4+CD25+ immunoregulatory T cells suppress polyclonal activation in vitro by inhibiting interleukin 2 production

    J Exp Med

    (1998)
  • S Read et al.

    Cytotoxic T lymphocyte-associated antigen 4 plays an essential role in the function of CD25(+)CD4(+) regulatory cells that control intestinal inflammation

    J Exp Med

    (2000)
  • X Zhang et al.

    Activation of CD25(+)CD4(+) regulatory T cells by oral antigen administration

    J Immunol

    (2001)
  • H Jiang et al.

    Role of CD8+ T cells in murine experimental allergic encephalomyelitis

    Science

    (1992)
  • Y Tada et al.

    Collagen-induced arthritis in CD4- or CD8-deficient mice: CD8+ T cells play a role in initiation and regulate recovery phase of collagen-induced arthritis

    J Immunol

    (1996)
  • L Zhang et al.

    Role of infused CD8+ cells in the induction of peripheral tolerance

    J Immunol

    (1994)
  • Cited by (0)

    *

    Address requests for reprints to: Matthieu Allez, M.D., Immunobiology Center, Mount Sinai School of Medicine, 1425 Madison Avenue, Room 11-20, New York, New York 10029. e-mail: [email protected]; fax: (212) 987-5593.

    **

    Supported by National Institutes of Health grants AI 23504, AI 24671, AI 44236, a Crohn's and Colitis Foundation of America fellowship award (to M.A. and I.D.), Société Nationale Française de Gastro-Entérologie (to M.A.), Institut de recherche des maladies de l'appareil digestif (to M.A.), Laboratoire Glaxo-Wellcome (to M.A.), and the Danish medical research agency (to J.B.).

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