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Developmental stage, phenotype, and migration distinguish naive- and effector/memory-like CD4+ regulatory T cells.

Huehn J, Siegmund K, Lehmann JC, Siewert C, Haubold U, Feuerer M, Debes GF, Lauber J, Frey O, Przybylski GK, Niesner U, de la Rosa M, Schmidt CA, Bräuer R, Buer J, Scheffold A, Hamann A - J. Exp. Med. (2004)

Bottom Line: We reported previously that the integrin alphaEbeta7 discriminates distinct subsets of murine CD4+ regulatory T cells.Use of this marker has now helped to unravel a fundamental dichotomy among regulatory T cells. alphaE-CD25+ cells expressed L-selectin and CCR7, enabling recirculation through lymphoid tissues.Accordingly, alphaE -expressing cells were found to be the most potent suppressors of inflammatory processes in disease models such as antigen-induced arthritis.

View Article: PubMed Central - PubMed

Affiliation: Experimentelle Rheumatologie, Medizinische Klinik, Charité, Humboldt-Universitaet, Schumannstr. 21/22, 10117 Berlin, Germany. Huehn@drfz.de

ABSTRACT
Regulatory T cells (Tregs) fulfill a central role in immune regulation. We reported previously that the integrin alphaEbeta7 discriminates distinct subsets of murine CD4+ regulatory T cells. Use of this marker has now helped to unravel a fundamental dichotomy among regulatory T cells. alphaE-CD25+ cells expressed L-selectin and CCR7, enabling recirculation through lymphoid tissues. In contrast, alphaE -positive subsets (CD25+ and CD25-) displayed an effector/memory phenotype expressing high levels of E/P-selectin-binding ligands, multiple adhesion molecules as well as receptors for inflammatory chemokines, allowing efficient migration into inflamed sites. Accordingly, alphaE -expressing cells were found to be the most potent suppressors of inflammatory processes in disease models such as antigen-induced arthritis.

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Foxp3 expression in Treg subsets. Foxp3 mRNA expression was determined in sorted Treg subsets by real-time quantitative RT-PCR. Obtained values were normalized to the housekeeping gene hypoxanthine phosphoribosyltransferase and expressed as percentage of the level found in CD25 single positive cells (mean ± SD of three independent experiments).
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fig1: Foxp3 expression in Treg subsets. Foxp3 mRNA expression was determined in sorted Treg subsets by real-time quantitative RT-PCR. Obtained values were normalized to the housekeeping gene hypoxanthine phosphoribosyltransferase and expressed as percentage of the level found in CD25 single positive cells (mean ± SD of three independent experiments).

Mentions: To obtain a comprehensive picture of the phenotype and developmental stage of αE-expressing Tregs in comparison to CD25 single positive cells, we applied cDNA microarray technology for gene expression profiling of αE−CD25+, αE+CD25+, and αE+CD25− CD4+ T cell subsets isolated directly ex vivo from secondary lymphoid organs. From ∼12,400 murine full-length genes and expressed sequence tags that were analyzed on Affymetrix microarrays, ∼90 (without expressed sequence tags) proved to be differentially expressed between the three Treg subsets when a ratio >2 in three independent experiments or confirmation by other techniques was presupposed (Fig. S2 available at http://www.jem.org/cgi/content/full/jem.20031562/DC1). Intriguingly, many molecules associated with effector/memory differentiation or homing mechanisms appeared to be differentially regulated between the Treg subsets. In general, the largest differences were found between αE−CD25+ and αE+CD25− cells, whereas the double positive αE+CD25+ subset often displayed intermediate levels of mRNA expression. A smaller number of genes was found to be expressed at higher levels in αE−CD25+ than in αE+CD25− cells, including CCR7 and L-selectin (CD62L) (2–5 fold difference), whereas the majority of genes showed higher levels in αE+CD25− than in αE−CD25+ cells, including molecules such as CD44, ICOS, granzyme B, FucTVII, integrin β1, CD54, CCR2, CXCR3, CCR6, and Ki67 (2–10 fold difference), which have been associated with activation and effector/memory status. Because the Affymetrix MG-U74Av2 microarray did not admit evaluation of Foxp3, we additionally performed a quantitative RT-PCR and observed a high Foxp3 expression level in all three Treg subsets in comparison to naive T cells (Fig. 1).


Developmental stage, phenotype, and migration distinguish naive- and effector/memory-like CD4+ regulatory T cells.

Huehn J, Siegmund K, Lehmann JC, Siewert C, Haubold U, Feuerer M, Debes GF, Lauber J, Frey O, Przybylski GK, Niesner U, de la Rosa M, Schmidt CA, Bräuer R, Buer J, Scheffold A, Hamann A - J. Exp. Med. (2004)

Foxp3 expression in Treg subsets. Foxp3 mRNA expression was determined in sorted Treg subsets by real-time quantitative RT-PCR. Obtained values were normalized to the housekeeping gene hypoxanthine phosphoribosyltransferase and expressed as percentage of the level found in CD25 single positive cells (mean ± SD of three independent experiments).
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2211798&req=5

fig1: Foxp3 expression in Treg subsets. Foxp3 mRNA expression was determined in sorted Treg subsets by real-time quantitative RT-PCR. Obtained values were normalized to the housekeeping gene hypoxanthine phosphoribosyltransferase and expressed as percentage of the level found in CD25 single positive cells (mean ± SD of three independent experiments).
Mentions: To obtain a comprehensive picture of the phenotype and developmental stage of αE-expressing Tregs in comparison to CD25 single positive cells, we applied cDNA microarray technology for gene expression profiling of αE−CD25+, αE+CD25+, and αE+CD25− CD4+ T cell subsets isolated directly ex vivo from secondary lymphoid organs. From ∼12,400 murine full-length genes and expressed sequence tags that were analyzed on Affymetrix microarrays, ∼90 (without expressed sequence tags) proved to be differentially expressed between the three Treg subsets when a ratio >2 in three independent experiments or confirmation by other techniques was presupposed (Fig. S2 available at http://www.jem.org/cgi/content/full/jem.20031562/DC1). Intriguingly, many molecules associated with effector/memory differentiation or homing mechanisms appeared to be differentially regulated between the Treg subsets. In general, the largest differences were found between αE−CD25+ and αE+CD25− cells, whereas the double positive αE+CD25+ subset often displayed intermediate levels of mRNA expression. A smaller number of genes was found to be expressed at higher levels in αE−CD25+ than in αE+CD25− cells, including CCR7 and L-selectin (CD62L) (2–5 fold difference), whereas the majority of genes showed higher levels in αE+CD25− than in αE−CD25+ cells, including molecules such as CD44, ICOS, granzyme B, FucTVII, integrin β1, CD54, CCR2, CXCR3, CCR6, and Ki67 (2–10 fold difference), which have been associated with activation and effector/memory status. Because the Affymetrix MG-U74Av2 microarray did not admit evaluation of Foxp3, we additionally performed a quantitative RT-PCR and observed a high Foxp3 expression level in all three Treg subsets in comparison to naive T cells (Fig. 1).

Bottom Line: We reported previously that the integrin alphaEbeta7 discriminates distinct subsets of murine CD4+ regulatory T cells.Use of this marker has now helped to unravel a fundamental dichotomy among regulatory T cells. alphaE-CD25+ cells expressed L-selectin and CCR7, enabling recirculation through lymphoid tissues.Accordingly, alphaE -expressing cells were found to be the most potent suppressors of inflammatory processes in disease models such as antigen-induced arthritis.

View Article: PubMed Central - PubMed

Affiliation: Experimentelle Rheumatologie, Medizinische Klinik, Charité, Humboldt-Universitaet, Schumannstr. 21/22, 10117 Berlin, Germany. Huehn@drfz.de

ABSTRACT
Regulatory T cells (Tregs) fulfill a central role in immune regulation. We reported previously that the integrin alphaEbeta7 discriminates distinct subsets of murine CD4+ regulatory T cells. Use of this marker has now helped to unravel a fundamental dichotomy among regulatory T cells. alphaE-CD25+ cells expressed L-selectin and CCR7, enabling recirculation through lymphoid tissues. In contrast, alphaE -positive subsets (CD25+ and CD25-) displayed an effector/memory phenotype expressing high levels of E/P-selectin-binding ligands, multiple adhesion molecules as well as receptors for inflammatory chemokines, allowing efficient migration into inflamed sites. Accordingly, alphaE -expressing cells were found to be the most potent suppressors of inflammatory processes in disease models such as antigen-induced arthritis.

Show MeSH
Related in: MedlinePlus