Expression of the integrin α_Eβ₇identifies unique subsets of CD25⁺as well as CD25⁻regulatory T cells

Abstract

Regulatory CD25 ؉ CD4 ؉ T cells are considered as important players in T cell homeostasis and self-tolerance. Here we report that the integrin ␣E␤7, which recognizes epithelial cadherin, identifies the most potent subpopulation of regulatory CD25 ؉ T cells. Strikingly, CD25-negative ␣E ؉ CD4 ؉ T cells displayed regulatory activity. Both ␣E ؉ subsets, CD25 ؉ and CD25 ؊ , express CTLA-4, suppress T cell proliferation in vitro, and protect mice from colitis in the severe combined immunodeficient model (SCID) in vivo. Whereas ␣E ؉ CD25 ؉ T cells produce almost no cytokines, ␣E ؉ CD25 ؊ T cells represent a unique subset in which high IL-2, IFN-␥ and T helper 2-cytokine production is linked with suppressive function. Thus, the integrin ␣E␤7 can be regarded as a novel marker for subsets of highly potent, functionally distinct regulatory T cells specialized for crosstalk with epithelial environments.

MHC class II‐independent CD25+ CD4+ CD8α β+ α β T cells attenuate CD4+ T cell‐induced transfer colitis

“…2). CD103 (integrin § E -chain), a marker of CD4 + T R cells [13] is expressed by 40-50% DP and 10-20% SP T cells while X 3% of CD4 + T cells in spleen, MLN or cLP of normal mice express this marker (Fig. 2).…”

“…In addition, other (CD25 + or CD25 -) CD4 + T R cell subsets have been shown to control transfer colitis. CD103 (integrin § E ) + (CD25 + or CD25 -) § g T R cells that express CTLA-4 protect mice from colitis pointing to CD103 as a marker for a T R cells subset apparently specialized for cross-talk with epithelial cells [13]. Activated, membrane TGF-g -expressing (CD25 + or CD25 -) T R cells suppress the manifestation of transfer colitis [14].…”

Induction of organ‐selective CD4+ regulatory T cell homing

“…CD103 -CD25 + CD4 + T cells were FACS-sorted using a FACS DIVA (BD, Heidelberg) and CFSE-labeled as described previously [54]. CFSE-labeled cells, 3 Â 10 5 -6 Â 10 5 in 200 lL PBS were injected i.v., and 20 h later, 500 lg OVA In vitro culture of Treg with organ-specific DC For isolation of organ-specific DC, meshed pLN or mLN from DO11.10 mice were incubated with 1 mg/mL collagenase D (Roche Diagnostics) and 20 lg/mL DNase (Sigma) for 30 min at 37 C. Subsequently, CD11c + cells were positively selected from single-cell suspensions using anti-CD11c microbeads and the AutoMACS magnetic separation system (Miltenyi Biotec).…”

“…CD103 -CD25 + CD4 + T cells were FACS-sorted using a FACS DIVA (BD, Heidelberg) and CFSE-labeled as described previously [54]. CFSE-labeled cells, 3 Â 10 5 -6 Â 10 5 in 200 lL PBS were injected i.v., and 20 h later, 500 lg OVA (Sigma) together with 2 lg cholera toxin (Sigma) or 70 lg LPS (Sigma) in 200 lL PBS were injected i.p.…”

“…For isolation of naïve-like CD4 + Treg, the positive fraction was stained with anti-CD25-FITC and CD62L-APC and CD25 + CD4 + CD62L high cells were sorted by FACS (DIVA or Aria, BD Bioscience) to a purity of >95%. FACS-sorted cells were labeled with CFSE as previously described [54], and 2 Â 10 5 isolated, CFSE-labeled Treg were stimulated for 4 days with 2 Â 10 4 CD11c + DC together with 1 lg/mL OVA 323-339 peptide (synthesized at the Dept. of Biochemistry, Humboldt-University, Berlin, Germany) and 40 ng/mL recombinant IL-2 (R&D Systems).…”