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TGF-β induces surface LAP expression on murine CD4 T cells independent of Foxp3 induction.

Oida T, Weiner HL - PLoS ONE (2010)

Bottom Line: It has been reported that human FOXP3(+) CD4 Tregs express GARP-anchored surface latency-associated peptide (LAP) after activation, based on the use of an anti-human LAP mAb.We then examined surface LAP expression after treating CD4(+)CD25(-) T cells with TGF-β and found that TGF-β induced surface LAP not only on T cells that became Foxp3(+) but also on T cells that remained Foxp3(-) after TGF-β treatment.Our newly described anti-mouse LAP mAbs will provide a useful tool for the investigation and functional analysis of T cells that express LAP on their surface.

View Article: PubMed Central - PubMed

Affiliation: Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.

ABSTRACT

Background: It has been reported that human FOXP3(+) CD4 Tregs express GARP-anchored surface latency-associated peptide (LAP) after activation, based on the use of an anti-human LAP mAb. Murine CD4 Foxp3(+) Tregs have also been reported to express surface LAP, but these studies have been hampered by the lack of suitable anti-mouse LAP mAbs.

Methodology/principal findings: We generated anti-mouse LAP mAbs by immunizing TGF-β(-/-) animals with a mouse Tgfb1-transduced P3U1 cell line. Using these antibodies, we demonstrated that murine Foxp3(+) CD4 Tregs express LAP on their surface. In addition, retroviral transduction of Foxp3 into mouse CD4(+)CD25(-) T cells induced surface LAP expression. We then examined surface LAP expression after treating CD4(+)CD25(-) T cells with TGF-β and found that TGF-β induced surface LAP not only on T cells that became Foxp3(+) but also on T cells that remained Foxp3(-) after TGF-β treatment. GARP expression correlated with the surface LAP expression, suggesting that surface LAP is GARP-anchored also in murine T cells.

Conclusions/significance: Unlike human CD4 T cells, surface LAP expression on mouse CD4 T cells is controlled by Foxp3 and TGF-β. Our newly described anti-mouse LAP mAbs will provide a useful tool for the investigation and functional analysis of T cells that express LAP on their surface.

Show MeSH
Induction of surface LAP by Foxp3 transduction.BALB/c CD4+CD25− T cells were stimulated with plate-bound anti-CD3/anti-CD28 and retrovirally transduced with pMCs-Foxp3-IRES-GFP vector. The cells were re-stimulated with plate-bound anti-CD3/anti-CD28 for 14 hrs and transferred to uncoated wells. 2 days after re-stimulation, the cells were stained with anti-LAP TW7-16B4 or TW7-20B9 using anti-mouse IgG1-APC secondary antibody.
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pone-0015523-g003: Induction of surface LAP by Foxp3 transduction.BALB/c CD4+CD25− T cells were stimulated with plate-bound anti-CD3/anti-CD28 and retrovirally transduced with pMCs-Foxp3-IRES-GFP vector. The cells were re-stimulated with plate-bound anti-CD3/anti-CD28 for 14 hrs and transferred to uncoated wells. 2 days after re-stimulation, the cells were stained with anti-LAP TW7-16B4 or TW7-20B9 using anti-mouse IgG1-APC secondary antibody.

Mentions: We then asked whether surface LAP expression is controlled by Foxp3. We found that retroviral Foxp3 transduction into mouse CD4+CD25− T cells induced surface LAP (GFP+ population vs. GFP− population in Figure 3). This result demonstrates that surface LAP is under control of Foxp3.


TGF-β induces surface LAP expression on murine CD4 T cells independent of Foxp3 induction.

Oida T, Weiner HL - PLoS ONE (2010)

Induction of surface LAP by Foxp3 transduction.BALB/c CD4+CD25− T cells were stimulated with plate-bound anti-CD3/anti-CD28 and retrovirally transduced with pMCs-Foxp3-IRES-GFP vector. The cells were re-stimulated with plate-bound anti-CD3/anti-CD28 for 14 hrs and transferred to uncoated wells. 2 days after re-stimulation, the cells were stained with anti-LAP TW7-16B4 or TW7-20B9 using anti-mouse IgG1-APC secondary antibody.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2991360&req=5

pone-0015523-g003: Induction of surface LAP by Foxp3 transduction.BALB/c CD4+CD25− T cells were stimulated with plate-bound anti-CD3/anti-CD28 and retrovirally transduced with pMCs-Foxp3-IRES-GFP vector. The cells were re-stimulated with plate-bound anti-CD3/anti-CD28 for 14 hrs and transferred to uncoated wells. 2 days after re-stimulation, the cells were stained with anti-LAP TW7-16B4 or TW7-20B9 using anti-mouse IgG1-APC secondary antibody.
Mentions: We then asked whether surface LAP expression is controlled by Foxp3. We found that retroviral Foxp3 transduction into mouse CD4+CD25− T cells induced surface LAP (GFP+ population vs. GFP− population in Figure 3). This result demonstrates that surface LAP is under control of Foxp3.

Bottom Line: It has been reported that human FOXP3(+) CD4 Tregs express GARP-anchored surface latency-associated peptide (LAP) after activation, based on the use of an anti-human LAP mAb.We then examined surface LAP expression after treating CD4(+)CD25(-) T cells with TGF-β and found that TGF-β induced surface LAP not only on T cells that became Foxp3(+) but also on T cells that remained Foxp3(-) after TGF-β treatment.Our newly described anti-mouse LAP mAbs will provide a useful tool for the investigation and functional analysis of T cells that express LAP on their surface.

View Article: PubMed Central - PubMed

Affiliation: Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.

ABSTRACT

Background: It has been reported that human FOXP3(+) CD4 Tregs express GARP-anchored surface latency-associated peptide (LAP) after activation, based on the use of an anti-human LAP mAb. Murine CD4 Foxp3(+) Tregs have also been reported to express surface LAP, but these studies have been hampered by the lack of suitable anti-mouse LAP mAbs.

Methodology/principal findings: We generated anti-mouse LAP mAbs by immunizing TGF-β(-/-) animals with a mouse Tgfb1-transduced P3U1 cell line. Using these antibodies, we demonstrated that murine Foxp3(+) CD4 Tregs express LAP on their surface. In addition, retroviral transduction of Foxp3 into mouse CD4(+)CD25(-) T cells induced surface LAP expression. We then examined surface LAP expression after treating CD4(+)CD25(-) T cells with TGF-β and found that TGF-β induced surface LAP not only on T cells that became Foxp3(+) but also on T cells that remained Foxp3(-) after TGF-β treatment. GARP expression correlated with the surface LAP expression, suggesting that surface LAP is GARP-anchored also in murine T cells.

Conclusions/significance: Unlike human CD4 T cells, surface LAP expression on mouse CD4 T cells is controlled by Foxp3 and TGF-β. Our newly described anti-mouse LAP mAbs will provide a useful tool for the investigation and functional analysis of T cells that express LAP on their surface.

Show MeSH