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Treadmill exercise induces murine cardiac allograft survival and generates regulatory T cell.

Uchiyama M, Jin X, Yin E, Shimokawa T, Niimi M - Transpl. Int. (2014)

Bottom Line: Adoptive transfer of whole splenocytes and CD4(+) cells from treadmill exercise recipients significantly prolonged allograft survival in naive secondary recipients (MSTs, 30 and 52 days, respectively), suggesting that regulatory cells was generated after treadmill exercise.Moreover, flow cytometry studies showed that CD4(+) CD25(+) Foxp3(+) cell population increased in treadmill exercise recipients.Therefore, postoperative but not pre-operative exercise could induce prolongation of survival of fully allogeneic cardiac allografts and generate CD4(+) CD25(+) Foxp3(+) regulatory T cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Surgery, Teikyo University, Tokyo, Japan; Department of Surgery, Teikyo University, Tokyo, Japan.

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Evidence of generation of regulatory cells in treadmill-exercised CBA allograft recipients. (a) Scheme on adoptive transfer study to confirm the generation of regulatory T cells. (b, c) Cardiac allograft survival after adoptive transfer of whole splenocytes (b) or CD4+ cells (c). (d–h) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from untreated mice and postoperative 1-week treadmill-exercised mice (d–g) and 100 days after adoptive transfer of CD4+ cell from longtime surviving secondary CBA recipients with B6 beating heart (h). Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal antibody or anti-Foxp3 polyclonal antibody. In (d–g), the left-hand panels show samples obtained from mice exercising on a treadmill, and the right-hand panels show samples from untreated mice (magnification ×40). In (h), all panels show samples obtained from longtime surviving transplant recipients in CD4+ cell adoptive transfer groups (magnification ×100) (i) CD4, CD25, and Foxp3 expression in splenocytes as determined by flow cytometry 1, 2, and 4 weeks after transplantation. The right-hand graph shows the percentage of CD4+CD25+Foxp3+ cells in the CD4+ cells as determined by flow cytometry. Data are mean ± SD values (n = 5 mice in each group). MST median survival time. *P < 0.05 and ###P < 0.001 for difference between two groups. NS not significant.
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fig02: Evidence of generation of regulatory cells in treadmill-exercised CBA allograft recipients. (a) Scheme on adoptive transfer study to confirm the generation of regulatory T cells. (b, c) Cardiac allograft survival after adoptive transfer of whole splenocytes (b) or CD4+ cells (c). (d–h) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from untreated mice and postoperative 1-week treadmill-exercised mice (d–g) and 100 days after adoptive transfer of CD4+ cell from longtime surviving secondary CBA recipients with B6 beating heart (h). Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal antibody or anti-Foxp3 polyclonal antibody. In (d–g), the left-hand panels show samples obtained from mice exercising on a treadmill, and the right-hand panels show samples from untreated mice (magnification ×40). In (h), all panels show samples obtained from longtime surviving transplant recipients in CD4+ cell adoptive transfer groups (magnification ×100) (i) CD4, CD25, and Foxp3 expression in splenocytes as determined by flow cytometry 1, 2, and 4 weeks after transplantation. The right-hand graph shows the percentage of CD4+CD25+Foxp3+ cells in the CD4+ cells as determined by flow cytometry. Data are mean ± SD values (n = 5 mice in each group). MST median survival time. *P < 0.05 and ###P < 0.001 for difference between two groups. NS not significant.

Mentions: Adoptive transfer studies were conducted to determine whether regulatory cells were generated in mice exercised on a treadmill. Thus, 30 days after transplantation of B6 hearts into primary CBA recipients exercised on a treadmill for 1 week after grafting, splenocytes (5.0 × 107) from primary recipients with functioning allografts were adoptively transferred into naïve secondary CBA recipients by means of intravenous injection into the penile vein. Immediately afterward, the secondary recipients underwent transplantation of a B6 heart. In some experiments, CD4+ cells were purified from the spleens of primary transplant recipients by positive selection using a magnetically activated cell sorter (MACS) and CD4 microbeads (Miltenyi Biotec, Auburn, CA; purity >98%), and CD4+ cells (2.0 × 107) were adoptively transferred into naïve secondary recipients, which then immediately underwent transplantation of a B6 heart (Fig.2a).


Treadmill exercise induces murine cardiac allograft survival and generates regulatory T cell.

Uchiyama M, Jin X, Yin E, Shimokawa T, Niimi M - Transpl. Int. (2014)

Evidence of generation of regulatory cells in treadmill-exercised CBA allograft recipients. (a) Scheme on adoptive transfer study to confirm the generation of regulatory T cells. (b, c) Cardiac allograft survival after adoptive transfer of whole splenocytes (b) or CD4+ cells (c). (d–h) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from untreated mice and postoperative 1-week treadmill-exercised mice (d–g) and 100 days after adoptive transfer of CD4+ cell from longtime surviving secondary CBA recipients with B6 beating heart (h). Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal antibody or anti-Foxp3 polyclonal antibody. In (d–g), the left-hand panels show samples obtained from mice exercising on a treadmill, and the right-hand panels show samples from untreated mice (magnification ×40). In (h), all panels show samples obtained from longtime surviving transplant recipients in CD4+ cell adoptive transfer groups (magnification ×100) (i) CD4, CD25, and Foxp3 expression in splenocytes as determined by flow cytometry 1, 2, and 4 weeks after transplantation. The right-hand graph shows the percentage of CD4+CD25+Foxp3+ cells in the CD4+ cells as determined by flow cytometry. Data are mean ± SD values (n = 5 mice in each group). MST median survival time. *P < 0.05 and ###P < 0.001 for difference between two groups. NS not significant.
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fig02: Evidence of generation of regulatory cells in treadmill-exercised CBA allograft recipients. (a) Scheme on adoptive transfer study to confirm the generation of regulatory T cells. (b, c) Cardiac allograft survival after adoptive transfer of whole splenocytes (b) or CD4+ cells (c). (d–h) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from untreated mice and postoperative 1-week treadmill-exercised mice (d–g) and 100 days after adoptive transfer of CD4+ cell from longtime surviving secondary CBA recipients with B6 beating heart (h). Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal antibody or anti-Foxp3 polyclonal antibody. In (d–g), the left-hand panels show samples obtained from mice exercising on a treadmill, and the right-hand panels show samples from untreated mice (magnification ×40). In (h), all panels show samples obtained from longtime surviving transplant recipients in CD4+ cell adoptive transfer groups (magnification ×100) (i) CD4, CD25, and Foxp3 expression in splenocytes as determined by flow cytometry 1, 2, and 4 weeks after transplantation. The right-hand graph shows the percentage of CD4+CD25+Foxp3+ cells in the CD4+ cells as determined by flow cytometry. Data are mean ± SD values (n = 5 mice in each group). MST median survival time. *P < 0.05 and ###P < 0.001 for difference between two groups. NS not significant.
Mentions: Adoptive transfer studies were conducted to determine whether regulatory cells were generated in mice exercised on a treadmill. Thus, 30 days after transplantation of B6 hearts into primary CBA recipients exercised on a treadmill for 1 week after grafting, splenocytes (5.0 × 107) from primary recipients with functioning allografts were adoptively transferred into naïve secondary CBA recipients by means of intravenous injection into the penile vein. Immediately afterward, the secondary recipients underwent transplantation of a B6 heart. In some experiments, CD4+ cells were purified from the spleens of primary transplant recipients by positive selection using a magnetically activated cell sorter (MACS) and CD4 microbeads (Miltenyi Biotec, Auburn, CA; purity >98%), and CD4+ cells (2.0 × 107) were adoptively transferred into naïve secondary recipients, which then immediately underwent transplantation of a B6 heart (Fig.2a).

Bottom Line: Adoptive transfer of whole splenocytes and CD4(+) cells from treadmill exercise recipients significantly prolonged allograft survival in naive secondary recipients (MSTs, 30 and 52 days, respectively), suggesting that regulatory cells was generated after treadmill exercise.Moreover, flow cytometry studies showed that CD4(+) CD25(+) Foxp3(+) cell population increased in treadmill exercise recipients.Therefore, postoperative but not pre-operative exercise could induce prolongation of survival of fully allogeneic cardiac allografts and generate CD4(+) CD25(+) Foxp3(+) regulatory T cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Surgery, Teikyo University, Tokyo, Japan; Department of Surgery, Teikyo University, Tokyo, Japan.

Show MeSH
Related in: MedlinePlus