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. 2000 Apr 17;191(8):1423-8.
doi: 10.1084/jem.191.8.1423.

Chronic restraint stress promotes lymphocyte apoptosis by modulating CD95 expression

D Yin  1 , D Tuthill, R A Mufson, Y Shi
Affiliations

Chronic restraint stress promotes lymphocyte apoptosis by modulating CD95 expression

D Yin et al. J Exp Med. .

Abstract

Depending on the duration and severity, psychological tension and physical stress can enhance or suppress the immune system in both humans and animals. Although it is well established that stress alters the release of various hormones and neurotransmitters, the mechanisms by which stress affects immune responses remain elusive. We report here that mice subjected to chronic 12-hour daily physical restraint for two days exhibited a significant reduction in splenocytes, a process likely mediated by apoptosis as demonstrated by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling assay. CD95 (Fas/APO-1) expression in splenic lymphocytes of stressed mice was substantially increased. Interestingly, Fas-immunoglobulin fusion protein and blocking antibodies against CD95 ligand inhibit stress-induced reduction in lymphocytes. The stress-induced changes in CD95 expression and lymphocyte number could be blocked by naltrexone or naloxone, specific opioid receptor antagonists, indicating a pivotal role of endogenous opioids in this process. In addition, the reduction of splenocytes in this model system seems to be independent of the hypothalamo-pituitary-adrenal axis, as both adrenalectomized and sham-operated mice exhibited similar responses to chronic stress. Moreover, chronic physical restraint failed to induce a decrease in lymphocyte numbers in CD95-deficient (Fas(lpr/lpr)) mice. Therefore, stress modulates the immune system through CD95-mediated apoptosis dependent on endogenous opioids.

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Figures

Figure 1
Figure 1
Physical restraint–induced lymphocyte reduction in the spleen requires opioid receptors and CD95–CD95L interaction. (A) Balb/c mice aged 7–9 wk were subjected to a 12-h physical restraint daily for 2 d. The following reagents were administered immediately before physical restraint in position: naltrexone (single intraperitoneal injection of 200 μg in PBS at 1 h before the initiation of each stress cycle), Fas-Ig (100 μg per mouse in PBS), and MFL3 (Anti-FasL; 10 μg per mouse in PBS). Mice were killed by CO2 asphyxiation, and total splenocytes were enumerated. Means and SEs were calculated from 5–10 mice per group. (B) Stress induces the expression of CD95. Balb/c mice at 7 wk of age were stressed for 12 h with or without anti–mouse FasL (10 μg per mouse) or naltrexone (200 μg per mouse) for 12 h. Total RNA was isolated from splenocytes and analyzed for CD95 expression by Northern blot hybridization with cDNA probe for CD95. GAPDH provides a control for RNA loading.
Figure 1
Figure 1
Physical restraint–induced lymphocyte reduction in the spleen requires opioid receptors and CD95–CD95L interaction. (A) Balb/c mice aged 7–9 wk were subjected to a 12-h physical restraint daily for 2 d. The following reagents were administered immediately before physical restraint in position: naltrexone (single intraperitoneal injection of 200 μg in PBS at 1 h before the initiation of each stress cycle), Fas-Ig (100 μg per mouse in PBS), and MFL3 (Anti-FasL; 10 μg per mouse in PBS). Mice were killed by CO2 asphyxiation, and total splenocytes were enumerated. Means and SEs were calculated from 5–10 mice per group. (B) Stress induces the expression of CD95. Balb/c mice at 7 wk of age were stressed for 12 h with or without anti–mouse FasL (10 μg per mouse) or naltrexone (200 μg per mouse) for 12 h. Total RNA was isolated from splenocytes and analyzed for CD95 expression by Northern blot hybridization with cDNA probe for CD95. GAPDH provides a control for RNA loading.
Figure 2
Figure 2
Physical restraint induces apoptotic cell death. (A) Balb/c 7–9-wk-old mice were subjected to a 12-h physical restraint daily for 2 d with or without anti–mouse FasL (10 μg per mouse) or naltrexone (200 μg per mouse). TUNEL-positive apoptotic cells in frozen spleen sections were shown in dark brown on a methyl green counterstained background. (B) Stress primes lymphocytes to CD95L-mediated apoptosis. Splenocytes from mice stressed for 12 h or from unstressed mice were cocultured with L cells transfected with CD95L cDNA in sense (S) or antisense (AS) orientations. Nonadherent cells were harvested at 24 h. DNA content was analyzed by flow cytometry after staining with propidium iodide.
Figure 2
Figure 2
Physical restraint induces apoptotic cell death. (A) Balb/c 7–9-wk-old mice were subjected to a 12-h physical restraint daily for 2 d with or without anti–mouse FasL (10 μg per mouse) or naltrexone (200 μg per mouse). TUNEL-positive apoptotic cells in frozen spleen sections were shown in dark brown on a methyl green counterstained background. (B) Stress primes lymphocytes to CD95L-mediated apoptosis. Splenocytes from mice stressed for 12 h or from unstressed mice were cocultured with L cells transfected with CD95L cDNA in sense (S) or antisense (AS) orientations. Nonadherent cells were harvested at 24 h. DNA content was analyzed by flow cytometry after staining with propidium iodide.
Figure 3
Figure 3
Mice bearing mutations in CD95 are resistant to physical restraint–induced lymphocyte reduction. C3H.MRL.Faslpr (C3H-lpr/lpr) or C3H/HeJ (C3H) mice aged 7–9 wk were subjected to a 12-h physical restraint daily for 3 d. Mice were killed by CO2 asphyxiation, and total splenocytes were enumerated. Means and SEs were calculated from five mice per group.
Figure 4
Figure 4
Adrenalectomy does not affect chronic restraint stress–induced splenocyte reduction. 10-wk-old Balb/c mice were adrenalectomized and subjected to physical restraint stress at 2–3-wk after operation. Both adrenalectomized mice and sham-operated mice were stressed for 12 h daily for 2 d. Mice were killed by CO2 asphyxiation at the end of the stress cycle, and total splenocytes were enumerated. The numbers of splenocytes were compared with the unstressed control. Cumulative means and SEs were calculated from six mice in two separate experiments.
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References

    1. Stone A.A., Bovbjerg D.H. Stress and humoral immunitya review of the human studies. Adv. Neuroimmunol. 1994;4:49–56. - PubMed
    1. Wilder R.L. Neuroendocrine-immune system interactions and autoimmunity. Annu. Rev. Immunol. 1995;13:307–338. - PubMed
    1. Gonzalez-Quijano M.I., Martin M., Millan S., Lopez-Calderon A. Lymphocyte response to mitogensinfluence of life events and personality. Neuropsychobiology. 1998;38:90–96. - PubMed
    1. Husband A.J. Role of central nervous system and behaviour in the immune response. Vaccine. 1993;11:805–816. - PubMed
    1. Pedersen B.K., Nieman D.C. Exercise immunologyintegration and regulation. Immunol. Today. 1998;19:204–206. - PubMed

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