Limits...
The Wiskott-Aldrich syndrome protein is required for the function of CD4(+)CD25(+)Foxp3(+) regulatory T cells.

Maillard MH, Cotta-de-Almeida V, Takeshima F, Nguyen DD, Michetti P, Nagler C, Bhan AK, Snapper SB - J. Exp. Med. (2007)

Bottom Line: Preincubation of WKO nTreg cells with exogenous interleukin (IL)-2, combined with antigen receptor-mediated activation, substantially rescues the suppression defects.WKO nTreg cells are also defective in the secretion of the immunomodulatory cytokine IL-10.Overall, our data reveal a critical role for WASP in nTreg cell function and implicate nTreg cell dysfunction in the autoimmunity associated with WASP deficiency.

View Article: PubMed Central - PubMed

Affiliation: Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, USA.

ABSTRACT
The Wiskott-Aldrich syndrome, a primary human immunodeficiency, results from defective expression of the hematopoietic-specific cytoskeletal regulator Wiskott-Aldrich syndrome protein (WASP). Because CD4(+)CD25(+)Foxp3(+) naturally occurring regulatory T (nTreg) cells control autoimmunity, we asked whether colitis in WASP knockout (WKO) mice is associated with aberrant development/function of nTreg cells. We show that WKO mice have decreased numbers of CD4(+)CD25(+)Foxp3(+) nTreg cells in both the thymus and peripheral lymphoid organs. Moreover, we demonstrate that WKO nTreg cells are markedly defective in both their ability to ameliorate the colitis induced by the transfer of CD45RB(hi) T cells and in functional suppression assays in vitro. Compared with wild-type (WT) nTreg cells, WKO nTreg cells show significantly impaired homing to both mucosal (mesenteric) and peripheral sites upon adoptive transfer into WT recipient mice. Suppression defects may be independent of antigen receptor-mediated actin rearrangement because both WT and WKO nTreg cells remodeled their actin cytoskeleton inefficiently upon T cell receptor stimulation. Preincubation of WKO nTreg cells with exogenous interleukin (IL)-2, combined with antigen receptor-mediated activation, substantially rescues the suppression defects. WKO nTreg cells are also defective in the secretion of the immunomodulatory cytokine IL-10. Overall, our data reveal a critical role for WASP in nTreg cell function and implicate nTreg cell dysfunction in the autoimmunity associated with WASP deficiency.

Show MeSH

Related in: MedlinePlus

WKO nTreg cells fail to suppress colitis induction in the CD45RB transfer model. (A) WT or WKO CD4+CD45RBhi T cells were injected alone or together with WT or WKO CD4+CD45RBlo T cells into SCID mice at day 0. Mice were assessed clinically weekly for a total of 8–12 wk followed by histological analysis. In a different set of experiments, CD4+CD25+ T cells were used instead of CD4+CD45RBlo T cells. (B) Clinical score of colitis 8 wk after transfer of WT or WKO CD4+CD45RBhi T cells and WT or WKO CD4+CD45RBlo T cells. Each dot represents an individual mouse. Horizontal bars represent average. Shown are combined data of four independent experiments. (C) Hematoxylin and eosin–stained cross sections of colonic sections taken 8 wk after transfer of WT CD4+CD45RBhi T cells and WT (left) or WKO (right) CD4+CD45RBlo T cells to SCID recipient mice. (D) Clinical score of colitis 8 wk after transfer of WT or WKO CD4+CD45RBhi T cells and WT or WKO CD4+CD25+ nTreg cells. Each dot represents an individual mouse. Horizontal bars represent average. Shown are the combined data of three independent experiments. NS, Nonsignificant (P > 0.05). *, P < 0.05; **, P < 0.01.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2118715&req=5

fig2: WKO nTreg cells fail to suppress colitis induction in the CD45RB transfer model. (A) WT or WKO CD4+CD45RBhi T cells were injected alone or together with WT or WKO CD4+CD45RBlo T cells into SCID mice at day 0. Mice were assessed clinically weekly for a total of 8–12 wk followed by histological analysis. In a different set of experiments, CD4+CD25+ T cells were used instead of CD4+CD45RBlo T cells. (B) Clinical score of colitis 8 wk after transfer of WT or WKO CD4+CD45RBhi T cells and WT or WKO CD4+CD45RBlo T cells. Each dot represents an individual mouse. Horizontal bars represent average. Shown are combined data of four independent experiments. (C) Hematoxylin and eosin–stained cross sections of colonic sections taken 8 wk after transfer of WT CD4+CD45RBhi T cells and WT (left) or WKO (right) CD4+CD45RBlo T cells to SCID recipient mice. (D) Clinical score of colitis 8 wk after transfer of WT or WKO CD4+CD45RBhi T cells and WT or WKO CD4+CD25+ nTreg cells. Each dot represents an individual mouse. Horizontal bars represent average. Shown are the combined data of three independent experiments. NS, Nonsignificant (P > 0.05). *, P < 0.05; **, P < 0.01.

Mentions: We first sought to determine whether WASP is required for nTreg cell suppression in vivo. To this end, we used a modification of the CD45RB transfer model of colitis (Fig. 2 A) (21). This model has the advantage of assessing both effector and regulatory T cell function. SCID mice were injected intraperitoneally with CD45RBhi T cells from either WT or WKO mice alone or in combination with WT or WKO CD45RBlo T cells (Fig. 2 A). Mice were assessed weekly for clinical signs of colitis for 8 wk, followed by histological analysis. WT CD45RBhi T cells, when injected alone, induced severe colitis (Fig. 2 B). Colitis induction was suppressed when WT CD45RBlo T cells were cotransferred with WT CD45RBhi T cells (Fig. 2, B and C). In contrast, WKO CD45RBlo T cells were unable to suppress colitis induction when cotransferred with WT CD45RBhi T cells (Fig. 2, B and C). When CD45RBhi T cells from WKO mice were injected alone, mice developed colitis, although associated with later onset and decreased severity (Fig. 2 B). Not surprisingly, fewer CD4+ T cells were recovered from the periphery of recipient mice receiving WKO CD45RBhi T cells (not depicted). Nonetheless, cotransfer of WKO CD45RBlo T cells could not prevent the onset of colitis in these mice. As expected, neither WT CD45RBlo nor WKO CD45RBlo T cells when injected alone induced colitis (Fig. 2 B). These data demonstrate that WKO CD45RBlo T cells were unable to protect against colitis induction by WT CD45RBhi T cells.


The Wiskott-Aldrich syndrome protein is required for the function of CD4(+)CD25(+)Foxp3(+) regulatory T cells.

Maillard MH, Cotta-de-Almeida V, Takeshima F, Nguyen DD, Michetti P, Nagler C, Bhan AK, Snapper SB - J. Exp. Med. (2007)

WKO nTreg cells fail to suppress colitis induction in the CD45RB transfer model. (A) WT or WKO CD4+CD45RBhi T cells were injected alone or together with WT or WKO CD4+CD45RBlo T cells into SCID mice at day 0. Mice were assessed clinically weekly for a total of 8–12 wk followed by histological analysis. In a different set of experiments, CD4+CD25+ T cells were used instead of CD4+CD45RBlo T cells. (B) Clinical score of colitis 8 wk after transfer of WT or WKO CD4+CD45RBhi T cells and WT or WKO CD4+CD45RBlo T cells. Each dot represents an individual mouse. Horizontal bars represent average. Shown are combined data of four independent experiments. (C) Hematoxylin and eosin–stained cross sections of colonic sections taken 8 wk after transfer of WT CD4+CD45RBhi T cells and WT (left) or WKO (right) CD4+CD45RBlo T cells to SCID recipient mice. (D) Clinical score of colitis 8 wk after transfer of WT or WKO CD4+CD45RBhi T cells and WT or WKO CD4+CD25+ nTreg cells. Each dot represents an individual mouse. Horizontal bars represent average. Shown are the combined data of three independent experiments. NS, Nonsignificant (P > 0.05). *, P < 0.05; **, P < 0.01.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2118715&req=5

fig2: WKO nTreg cells fail to suppress colitis induction in the CD45RB transfer model. (A) WT or WKO CD4+CD45RBhi T cells were injected alone or together with WT or WKO CD4+CD45RBlo T cells into SCID mice at day 0. Mice were assessed clinically weekly for a total of 8–12 wk followed by histological analysis. In a different set of experiments, CD4+CD25+ T cells were used instead of CD4+CD45RBlo T cells. (B) Clinical score of colitis 8 wk after transfer of WT or WKO CD4+CD45RBhi T cells and WT or WKO CD4+CD45RBlo T cells. Each dot represents an individual mouse. Horizontal bars represent average. Shown are combined data of four independent experiments. (C) Hematoxylin and eosin–stained cross sections of colonic sections taken 8 wk after transfer of WT CD4+CD45RBhi T cells and WT (left) or WKO (right) CD4+CD45RBlo T cells to SCID recipient mice. (D) Clinical score of colitis 8 wk after transfer of WT or WKO CD4+CD45RBhi T cells and WT or WKO CD4+CD25+ nTreg cells. Each dot represents an individual mouse. Horizontal bars represent average. Shown are the combined data of three independent experiments. NS, Nonsignificant (P > 0.05). *, P < 0.05; **, P < 0.01.
Mentions: We first sought to determine whether WASP is required for nTreg cell suppression in vivo. To this end, we used a modification of the CD45RB transfer model of colitis (Fig. 2 A) (21). This model has the advantage of assessing both effector and regulatory T cell function. SCID mice were injected intraperitoneally with CD45RBhi T cells from either WT or WKO mice alone or in combination with WT or WKO CD45RBlo T cells (Fig. 2 A). Mice were assessed weekly for clinical signs of colitis for 8 wk, followed by histological analysis. WT CD45RBhi T cells, when injected alone, induced severe colitis (Fig. 2 B). Colitis induction was suppressed when WT CD45RBlo T cells were cotransferred with WT CD45RBhi T cells (Fig. 2, B and C). In contrast, WKO CD45RBlo T cells were unable to suppress colitis induction when cotransferred with WT CD45RBhi T cells (Fig. 2, B and C). When CD45RBhi T cells from WKO mice were injected alone, mice developed colitis, although associated with later onset and decreased severity (Fig. 2 B). Not surprisingly, fewer CD4+ T cells were recovered from the periphery of recipient mice receiving WKO CD45RBhi T cells (not depicted). Nonetheless, cotransfer of WKO CD45RBlo T cells could not prevent the onset of colitis in these mice. As expected, neither WT CD45RBlo nor WKO CD45RBlo T cells when injected alone induced colitis (Fig. 2 B). These data demonstrate that WKO CD45RBlo T cells were unable to protect against colitis induction by WT CD45RBhi T cells.

Bottom Line: Preincubation of WKO nTreg cells with exogenous interleukin (IL)-2, combined with antigen receptor-mediated activation, substantially rescues the suppression defects.WKO nTreg cells are also defective in the secretion of the immunomodulatory cytokine IL-10.Overall, our data reveal a critical role for WASP in nTreg cell function and implicate nTreg cell dysfunction in the autoimmunity associated with WASP deficiency.

View Article: PubMed Central - PubMed

Affiliation: Gastrointestinal Unit, Massachusetts General Hospital, Boston, MA 02114, USA.

ABSTRACT
The Wiskott-Aldrich syndrome, a primary human immunodeficiency, results from defective expression of the hematopoietic-specific cytoskeletal regulator Wiskott-Aldrich syndrome protein (WASP). Because CD4(+)CD25(+)Foxp3(+) naturally occurring regulatory T (nTreg) cells control autoimmunity, we asked whether colitis in WASP knockout (WKO) mice is associated with aberrant development/function of nTreg cells. We show that WKO mice have decreased numbers of CD4(+)CD25(+)Foxp3(+) nTreg cells in both the thymus and peripheral lymphoid organs. Moreover, we demonstrate that WKO nTreg cells are markedly defective in both their ability to ameliorate the colitis induced by the transfer of CD45RB(hi) T cells and in functional suppression assays in vitro. Compared with wild-type (WT) nTreg cells, WKO nTreg cells show significantly impaired homing to both mucosal (mesenteric) and peripheral sites upon adoptive transfer into WT recipient mice. Suppression defects may be independent of antigen receptor-mediated actin rearrangement because both WT and WKO nTreg cells remodeled their actin cytoskeleton inefficiently upon T cell receptor stimulation. Preincubation of WKO nTreg cells with exogenous interleukin (IL)-2, combined with antigen receptor-mediated activation, substantially rescues the suppression defects. WKO nTreg cells are also defective in the secretion of the immunomodulatory cytokine IL-10. Overall, our data reveal a critical role for WASP in nTreg cell function and implicate nTreg cell dysfunction in the autoimmunity associated with WASP deficiency.

Show MeSH
Related in: MedlinePlus