Sleep-dependent activity of T cells and regulatory T cells - PubMed (original) (raw)

Sleep-dependent activity of T cells and regulatory T cells

T Bollinger et al. Clin Exp Immunol. 2009 Feb.

Abstract

A number of immunological functions are dependent on circadian rhythms and regular sleep. This has impact on the type and magnitude of immune responses following antigenic challenge, for example in vaccination. Little is known about the underlying mechanisms. One possibility may be the circadian and sleep-dependent modulation of CD4(+)CD25(-) T cell responses by CD4(+)CD25(+) natural regulatory T cells (nT(reg)). In a variety of studies, nT(reg) have been shown to regulate T cell responses negatively. Thus, we investigated the influence of sleep and circadian rhythm on the number and function of nT(reg) as well as on the function of CD4(+)CD25(-) T cells. Seven healthy young men were examined under defined conditions on two occasions, i.e. during sleep and sleep deprivation. Venous blood was drawn periodically; numbers of nT(reg), suppressive activity of nT(reg), interleukin-2 production and proliferation of CD4(+)CD25(-) T cells were explored in vitro. nT(reg) counts revealed a significant circadian rhythm with highest levels during the night (mean 95 nT(reg)/microl) and lowest levels during the day (mean 55 nT(reg)/microl). During normal sleep, the suppressive activity of nT(reg) was highest at 02.00 h and somewhat lower at 15.00 h. Surprisingly, almost no suppressive activity was present at 07.00 h. Deprivation of sleep abrogated this rhythm. CD4(+)CD25(-) T cell proliferation was dampened significantly by sleep deprivation. This is the first study in human cells to show that nT(reg) number and function follow a rhythm across the 24-h period. Furthermore, sleep deprivation severely disturbs the functional rhythm of nT(reg) and CD4(+)CD25(-) T cells.

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Figures

Fig. 1

Fig. 1

T cell and natural T regulatory (nTreg) cell purity and fluorescence activated cell sorter (FACS) analysis of CD4+CD25− T cell proliferation. (a) CD4+CD25− T cells (left panel) and CD4+CD25+ regulatory T cells (middle and right panel) were isolated from peripheral blood mononuclear cells (PBMC) applying magnetic affinity cell sorter (MACS®) technology. Purified T cells were stained with αCD4-monoclonal antibody (mAb) labelled with allophycocyanin and αCD25-mAb labelled with phycoerythrin or with αCD4-mAb labelled with fluorescein and αforkhead box P3 (FoxP3)-mAb labelled with allophycocyanin and were then analysed by flow cytometry. Mean purity of CD4+CD25− T cells was 96·8% ± 0·6% and mean purity of CD4+CD25+ nTreg was 79·4% ± 7·1%. (b) CD4+CD25−carboxyfluorescein diacetate (CFDA)+ T cells were cultured with nTreg (middle panel) or without nTreg (left panel) in the presence of irradiated adherent cells and αCD3-mAb. Proliferation of CD4+CD25−CFDA+ T cells was measured as the reduction in CFDA fluorescence. M2 represents the percentage of proliferated CD4+CD25− T cells. Control cultures were performed without αCD3-mAb (right panel). Data are from one representative experiment of 70.

Fig. 2

Fig. 2

Absolute counts of natural T regulatory (nTreg) and suppression of CD4+ T cell proliferation through nTreg. (a) To analyse the number of CD4+CD25+ regulatory T cells in peripheral blood we quantified the CD4+CD25high cells indicated by the circle. The percentage of forkhead box P3+ (FoxP3+) cells within the CD4+CD25high population did not differ over the 24-h period or between the sleep and sleep deprivation condition; 91·5% ± 1% of the CD4+CD25high cells were positive for FoxP3 (data not shown). (b) CD4+CD25+ regulatory T cells were counted in peripheral blood from healthy men during sleep (closed circles) or sleep deprivation (open circles). (c) CD4+CD25− T cells and nTreg were isolated from peripheral blood of healthy young men with sleep (closed circles) and without nocturnal sleep (open circles). CD4+CD25− T cells were stimulated polyclonally either in the presence or absence of nTreg. The inhibition of CD4+CD25− T cell proliferation by nTreg was calculated (for detailed information see Material and methods). (d) Peripheral blood mononuclear cells (PBMC) and PBMC depleted of nTreg were stimulated polyclonally and the inhibition of CD4+ T cell proliferation through the presence of nTreg was quantified in cells isolated from healthy young men with sleep (closed circles) or sleep deprivation (open circles). Shaded area indicates bedtime. Mean values ± standard error of the mean (n = 7).

Fig. 3

Fig. 3

Proliferation and interleukin (IL)-2 production of CD4+CD25− T cells in vitro and forkhead box P3 (FoxP3) expression in natural T regulatory (nTreg) cells. (a) CD4+CD25− T cells were isolated from peripheral blood of healthy young men with sleep (closed circles) or sleep deprivation (open circles), labelled with carboxyfluorescein diacetate (CFDA) and polyclonally stimulated. The percentage of proliferated T cell was measured through the reduction in CFDA fluorescence. (b, d) IL-2 was measured in the supernatants of polyclonally stimulated CD4+CD25− T cells in the absence (b) or presence (d) of nTreg. (c) FoxP3 expression levels in nTreg (CD4+CD25high) were measured as ‘Geo Mean’ applying flow cytometry. Cells were isolated from peripheral blood of healthy young men with sleep or sleep deprivation. Shaded area indicates bedtime. Mean values ± standard error of the mean (n = 7).

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