Tolerance induced by inhaled antigen involves CD4+ T cells expressing membrane-bound TGF-β and FOXP3
Marina Ostroukhova, … , Timothy E. Corcoran, Anuradha Ray
Marina Ostroukhova, … , Timothy E. Corcoran, Anuradha Ray
Published July 1, 2004
Citation Information: J Clin Invest. 2004;114(1):28-38. https://doi.org/10.1172/JCI20509.
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Article Immunology

Tolerance induced by inhaled antigen involves CD4+ T cells expressing membrane-bound TGF-β and FOXP3

Abstract

Under normal circumstances, the respiratory tract maintains immune tolerance in the face of constant antigen provocation. Using a murine model of tolerance induced by repeated exposure to a low dose of aerosolized antigen, we show an important contribution by CD4+ T cells in the establishment and maintenance of tolerance. The CD4+ T cells expressed both cell surface and soluble TGF-β and inhibited the development of an allergic phenotype when adoptively transferred to naive recipient mice. While cells expressing cell surface TGF-β were detectable in mice with inflammation, albeit at a lower frequency compared with that in tolerized mice, only those from tolerized mice expressed FOXP3. Blockade of TGF-β in vitro and in vivo interfered with immunosuppression. Although cells that expressed TGF-β on the cell surface (TGF-β+), as well as the ones that did not (TGF-β–), secreted equivalent levels of soluble TGF-β, only the former were able to blunt the development of an allergic phenotype in mice. Strikingly, separation of the TGF-β+ cells from the rest of the cells allowed the TGF-β– cells to proliferate in response to antigen. We propose a model of antigen-induced tolerance that involves cell-cell contact with regulatory CD4+ T cells that coexpress membrane-bound TGF-β and FOXP3.

Authors

Marina Ostroukhova, Carole Seguin-Devaux, Timothy B. Oriss, Barbara Dixon-McCarthy, Liyan Yang, Bill T. Ameredes, Timothy E. Corcoran, Anuradha Ray

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Figure 4

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Cell contact and TGF-β–dependent inhibition of proliferation by CD4+ T c...
Cell contact and TGF-β–dependent inhibition of proliferation by CD4+ T cells from tolerized mice. (A) Mice were first exposed to PBS (inflammation group) or 1% OVA (tolerance group) daily for 10 days and then were immunized with OVA/alum on days 21 and 27. Splenic CD4+ T cells isolated on day 34 were stimulated in vitro with different concentrations of OVA (10–200 ∝g/ml) and APCs at equivalent cell numbers (105 cells per well). Cells were mixed as described in the legend to Figure 2 or separated by transwell. In the transwell experiments, cells from the inflammation group were plated in the wells, and cells from tolerized mice on the insert and thymidine incorporation in the former group was measured. *P < 0.05 versus proliferation of cells in the inflammation group. (B) Chicken IgY anti–TGF-β1 (100 ng/ml) or isotype control (chicken IgY) was added to mixed cultures. **P < 0.05 of mixed cultures incubated with anti–TGF-β1 compared with mixed cultures incubated without Ab. (C) Anti–IL-10 (1 mg/ml) or isotype control was added to mixed cultures. All assays were incubated for 72 hours, after which the cells were pulsed for measurement of [3H]-thymidine incorporation. Each data point represents the mean plus or minus SEM of triplicate wells. Shown is a representative experiment of three experiments.