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Takeshi Kezuka, J. Wayne Streilein; Analysis of In Vivo Regulatory Properties of T Cells Activated In Vitro by TGFβ2-Treated Antigen Presenting Cells. Invest. Ophthalmol. Vis. Sci. 2000;41(6):1410-1421.
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purpose. To determine whether naive T cells activated in vitro by
antigen-pulsed, transforming growth factorβ (TGFβ)–treated antigen
presenting cells (APCs) acquire the capacity to suppress the induction
and expression of delayed hypersensitivity in vivo.
methods. Naive ovalbumin (OVA)-specific T cells from DO11.10 Tcr transgenic mice
were stimulated in vitro with OVA-pulsed TGFβ2-treated APCs. The
cultured cells were harvested and assayed for in vitro production of
mature TGFβ. Similar cells were coinjected with primed OVA-specific
BALB/c T cells plus OVA-pulsed APCs into ear pinnae of normal BALB/c
mice (assay for delayed hypersensitivity expression) or coinjected with
OVA-pulsed APCs into footpads of naive DO11.10 mice whose draining
lymph node cells were harvested 4 days later and assayed in vitro for
capacity to secrete interferon-γ (IFN-γ) and interleukin-4 (IL-4)
when stimulated with OVA (assay for induction of delayed
results. DO11.10 T cells activated in vitro by OVA-pulsed TGFβ2-treated APCs
secreted large amounts of mature TGFβ and suppressed the expression
of delayed hypersensitivity in a local adoptive transfer assay.
Suppression was reversed in the presence of neutralizing anti-TGFβ
antibodies. In addition, in vitro generated regulatory T cells
influenced naive T cells in DO11.10 mice that were responding to an
initial immunization with OVA to secrete IL-4, rather than IFN-γ.
This influence was independent of TGFβ.
conclusions. OVA-pulsed APCs, pretreated in vitro with TGFβ2, activate DO11.10 T
cells in a manner that endows the responding cells with the capacity to
suppress the induction and then the expression of delayed
hypersensitivity in vivo. In certain ways, these properties of in
vitro–activated DO11.10 T cells resemble the properties of afferent
and efferent regulatory T cells typically found in the spleens of
animals with anterior chamber–associated immune
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