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Phenotypic and functional characterization of a CD4 + CD25 high FOXP3 high regulatory T‐cell population in the dog
Author(s) -
Pinheiro Dammy,
Singh Yogesh,
Grant Charlotte R.,
Appleton Richard C.,
Sacchini Flavio,
Walker Kate R. L.,
Chadbourne Alden H.,
Palmer Charlotte A.,
ArmitageChan Elizabeth,
Thompson Ian,
Williamson Lina,
Cunningham Fiona,
Garden Oliver A.
Publication year - 2011
Publication title -
immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.297
H-Index - 133
eISSN - 1365-2567
pISSN - 0019-2805
DOI - 10.1111/j.1365-2567.2010.03346.x
Subject(s) - foxp3 , il 2 receptor , biology , interleukin 21 , population , clone (java method) , microbiology and biotechnology , cytotoxic t cell , interleukin 3 , cd8 , cd40 , peripheral blood mononuclear cell , t cell , interleukin 12 , immunology , in vitro , immune system , medicine , biochemistry , gene , environmental health
Summary Relatively little is known about regulatory T (Treg) cells and their functional responses in dogs. We have used the cross‐reactive anti‐mouse/rat Foxp3 antibody clone FJK‐16s to identify a population of canine CD4 + FOXP3 high T cells in both the peripheral blood (PB) and popliteal lymph node (LN). FOXP3 + cells in both PB and LN yielded positive staining with the newly developed anti‐murine/human Helios antibody clone 22F6, consistent with the notion that they were naturally occurring Treg cells. Stimulation of mononuclear cells of LN origin with concanavalin A (Con A) in vitro yielded increased proportions and median fluorescence intensity of FOXP3 expression by both CD4 + and CD8 + T cells. Removal of the Con A and continued culture disclosed a CD4 + FOXP3 high population, distinct from the CD4 + FOXP3 intermediate T cells; very few CD8 + FOXP3 high T cells were observed, though CD8 + FOXP3 intermediate cells were present in equal abundance to CD4 + FOXP3 intermediate cells. The CD4 + FOXP3 high T cells were thought to represent activated Treg cells, in contrast to the FOXP3 intermediate cells, which were thought to be a more heterogeneous population comprising predominantly activated conventional T cells. Co‐staining with interferon‐γ (IFN‐γ) supported this notion, because the FOXP3 high T cells were almost exclusively IFN‐γ − , whereas the FOXP3 intermediate cells expressed a more heterogeneous IFN‐γ phenotype. Following activation of mononuclear cells with Con A and interleukin‐2, the 5% of CD4 + T cells showing the highest CD25 expression (CD4 + CD25 high ) were enriched in cells expressing FOXP3. These cells were anergic in vitro , in contrast to the 20% of CD4 + T cells with the lowest CD25 expression (CD4 + CD25 − ), which proliferated readily. The CD4 + CD25 high FOXP3 high T cells were able to suppress the proliferation of responder CD4 + T cells in vitro , in contrast to the CD4 + CD25 − cells, which showed no regulatory properties.