Molecular Medicine

, Volume 20, Issue 1, pp 179–190 | Cite as

Environmental Enrichment Alters Splenic Immune Cell Composition and Enhances Secondary Influenza Vaccine Responses in Mice

  • Blake T. Gurfein
  • Olga Davidenko
  • Mary Premenko-Lanier
  • Jeffrey M. Milush
  • Michael Acree
  • Mary F. Dallman
  • Chadi Touma
  • Rupert Palme
  • Vanessa A. York
  • Gilles Fromentin
  • Nicolas Darcel
  • Douglas F. Nixon
  • Frederick M. Hecht
Research Article


Chronic stress has deleterious effects on immune function, which can lead to adverse health outcomes. However, studies investigating the impact of stress reduction interventions on immunity in clinical research have yielded divergent results, potentially stemming from differences in study design and genetic heterogeneity, among other clinical research challenges. To test the hypothesis that reducing glucocorticoid levels enhances certain immune functions, we administered influenza vaccine once (prime) or twice (boost) to mice housed in either standard control caging or environmental enrichment (EE) caging. We have shown that this approach reduces mouse corticosterone production. Compared with controls, EE mice had significantly lower levels of fecal corticosterone metabolites (FCMs) and increased splenic B and T lymphocyte numbers. Corticosterone levels were negatively associated with the numbers of CD19+ (r2 = 0.43, p = 0.0017), CD4+ (r2 = 0.28, p = 0.0154) and CD8+ cells (r2 = 0.20, p = 0.0503). Vaccinated mice showed nonsignificant differences in immunoglobulin G (IgG) titer between caging groups, although EE mice tended to exhibit larger increases in titer from prime to boost than controls; the interaction between the caging group (control versus EE) and vaccine group (prime versus boost) showed a strong statistical trend (cage-group*vaccine-group, F = 4.27, p = 0.0555), suggesting that there may be distinct effects of EE caging on primary versus secondary IgG vaccine responses. Vaccine-stimulated splenocytes from boosted EE mice had a significantly greater frequency of interleukin 5 (IL-5)-secreting cells than boosted controls (mean difference 7.7, IL-5 spot-forming units/106 splenocytes, 95% confidence interval 0.24–135.1, p = 0.0493) and showed a greater increase in the frequency of IL-5-secreting cells from prime to boost. Our results suggest that corticosterone reduction via EE caging was associated with enhanced secondary vaccine responses, but had little effect on primary responses in mice. These findings help identify differences in primary and secondary vaccine responses in relationship to stress mediators that may be relevant in clinical studies.


  1. 1.
    Lazarus RS, Folkman S. (1984) Stress, Appraisal, and Coping. Springer, New York.Google Scholar
  2. 2.
    Glaser R, Kiecolt-Glaser JK. (2005) Stress-induced immune dysfunction: implications for health. Nat. Rev. Immunol. 5:243–51.CrossRefPubMedGoogle Scholar
  3. 3.
    Edwards KM, et al. (2006) Acute stress exposure prior to influenza vaccination enhances antibody response in women. Brain Behav. Immun. 20:159–68.CrossRefPubMedGoogle Scholar
  4. 4.
    Schedlowski M, et al. (1993) Changes of natural killer cells during acute psychological stress. J. Clin. Immunol. 13:119–26.CrossRefPubMedGoogle Scholar
  5. 5.
    Dhabhar FS, McEwen BS. (1997) Acute stress enhances while chronic stress suppresses cell-mediated immunity in vivo: a potential role for leukocyte trafficking. Brain Behav. Immun. 11:286–306.CrossRefPubMedGoogle Scholar
  6. 6.
    Yin D, Tuthill D, Mufson RA, Shi Y. (2000) Chronic restraint stress promotes lymphocyte apoptosis by modulating CD95 expression. J. Exp. Med. 191:1423–8.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Thaker PH, et al. (2006) Chronic stress promotes tumor growth and angiogenesis in a mouse model of ovarian carcinoma. Nat. Med. 12:939–44.CrossRefPubMedGoogle Scholar
  8. 8.
    Kiecolt-Glaser JK, Marucha PT, Malarkey WB, Mercado AM, Glaser R. (1995) Slowing of wound healing by psychological stress. Lancet. 346:1194–6.CrossRefGoogle Scholar
  9. 9.
    Cohen S, Tyrrell DA, Smith AP. (1991) Psychological stress and susceptibility to the common cold. N. Engl. J. Med. 325:606–12.CrossRefPubMedGoogle Scholar
  10. 10.
    Cohen S, Doyle WJ, Skoner DP. (1999) Psychological stress, cytokine production, and severity of upper respiratory illness. Psychosom. Med. 61:175–80.CrossRefPubMedGoogle Scholar
  11. 11.
    Gallagher S, Phillips AC, Drayson MT, Carroll D. (2009) Parental caregivers of children with developmental disabilities mount a poor antibody response to pneumococcal vaccination. Brain Behav. Immun. 23:338–46.CrossRefPubMedGoogle Scholar
  12. 12.
    Vedhara K, et al. (1999) Chronic stress in elderly carers of dementia patients and antibody response to influenza vaccination. Lancet. 353:627–31.CrossRefPubMedGoogle Scholar
  13. 13.
    Glaser R, et al. (1992) Stress-induced modulation of the immune response to recombinant hepatitis B vaccine. Psychosom. Med. 54:22–9.CrossRefPubMedGoogle Scholar
  14. 14.
    Pedersen AF, Zachariae R, Bovbjerg DH. (2009) Psychological stress and antibody response to influenza vaccination: a meta-analysis. Brain Behav. Immun. 23:427–33.CrossRefPubMedGoogle Scholar
  15. 15.
    Segerstrom SC, Miller GE. (2004) Psychological stress and the human immune system: a meta-analytic study of 30 years of inquiry. Psychol. Bull. 130:601–30.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Carlson LE, Speca M, Faris P, Patel KD. (2007) One year pre-post intervention follow-up of psychological, immune, endocrine and blood pressure outcomes of mindfulness-based stress reduction (MBSR) in breast and prostate cancer outpatients. Brain Behav. Immun. 21:1038–49.CrossRefPubMedGoogle Scholar
  17. 17.
    Jain S, et al. (2007) A randomized controlled trial of mindfulness meditation versus relaxation training: effects on distress, positive states of mind, rumination, and distraction. Ann. Behav. Med. 33:11–21.CrossRefPubMedGoogle Scholar
  18. 18.
    Kabat-Zinn J, et al. (1992) Effectiveness of a meditation-based stress reduction program in the treatment of anxiety disorders. Am. J. Psychiatry. 149:936–43.CrossRefPubMedGoogle Scholar
  19. 19.
    Miller JJ, Fletcher K, Kabat-Zinn J. (1995) Three-year follow-up and clinical implications of a mindfulness meditation-based stress reduction intervention in the treatment of anxiety disorders. Gen. Hosp. Psychiatry. 17:192–200.CrossRefPubMedGoogle Scholar
  20. 20.
    Speca M, Carlson LE, Goodey E, Angen M. (2000) A randomized, wait-list controlled clinical trial: the effect of a mindfulness meditation-based stress reduction program on mood and symptoms of stress in cancer outpatients. Psychosom. Med. 62:613–22.CrossRefPubMedGoogle Scholar
  21. 21.
    Witek-Janusek L, et al. (2008) Effect of mindfulness based stress reduction on immune function, quality of life and coping in women newly diagnosed with early stage breast cancer. Brain. Behav. Immun. 22:969–81.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Hayney MS, et al. (2014) Age and psychological influences on immune responses to trivalent inactivated influenza vaccine in the meditation or exercise for preventing acute respiratory infection (MEPARI) trial. Hum. Vaccin. Immunother. 10:83–91.CrossRefPubMedGoogle Scholar
  23. 23.
    Davidson RJ, et al. (2003) Alterations in brain and immune function produced by mindfulness meditation. Psychosom. Med. 65:564–70.CrossRefPubMedGoogle Scholar
  24. 24.
    Moynihan JA, et al. (2013) Mindfulness-based stress reduction for older adults: effects on executive function, frontal alpha asymmetry and immune function. Neuropsychobiology. 68:34–43.CrossRefPubMedGoogle Scholar
  25. 25.
    Gurfein BT, et al. (2012) The calm mouse: an animal model of stress reduction. Mol. Med. 18:606–17.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Palme R, Touma C, Arias N, Dominchin MF, Lepschy M. (2013) Steroid extraction: get the best out of faecal samples. Wiener Tiera?rztliche Monatsschrift. 100:238–46.Google Scholar
  27. 27.
    Touma C, Sachser N, Mostl E, Palme R. (2003) Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice. Gen. Comp. Endocrinol. 130:267–78.CrossRefPubMedGoogle Scholar
  28. 28.
    Touma C, Palme R. (2005) Measuring fecal glucocorticoid metabolites in mammals and birds: the importance of validation. Ann. N. Y. Acad. Sci. 1046:54–74.CrossRefPubMedGoogle Scholar
  29. 29.
    Touma C, Palme R, Sachser N. (2004) Analyzing corticosterone metabolites in fecal samples of mice: a noninvasive technique to monitor stress hormones. Horm. Behav. 45:10–22.CrossRefPubMedGoogle Scholar
  30. 30.
    Louch CD, Higginbotham M. (1967) The relation between social rank and plasma corticosterone levels in mice. Gen. Comp. Endocrinol. 8:441–4.CrossRefPubMedGoogle Scholar
  31. 31.
    Gessner A, Blum H, Rollinghoff M. (1993) Differential regulation of IL-9-expression after infection with Leishmania major in susceptible and resistant mice. Immunobiology. 189:419–35.CrossRefPubMedGoogle Scholar
  32. 32.
    Locksley RM, Heinzel FP, Sadick MD, Holaday BJ, Gardner KD, Jr. (1987) Murine cutaneous leishmaniasis: susceptibility correlates with differential expansion of helper T-cell subsets. Ann. Inst. Pasteur. Immunol. 138:744–49.CrossRefPubMedGoogle Scholar
  33. 33.
    Moon BG, Takaki S, Miyake K, Takatsu K. (2004) The role of IL-5 for mature B-1 cells in homeostatic proliferation, cell survival, and Ig production. J. Immunol. 172:6020–9.CrossRefPubMedGoogle Scholar
  34. 34.
    Cao L, et al. (2010) Environmental and genetic activation of a brain-adipocyte BDNF/leptin axis causes cancer remission and inhibition. Cell. 142:52–64.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Benaroya-Milshtein N, et al. (2004) Environmental enrichment in mice decreases anxiety, attenuates stress responses and enhances natural killer cell activity. Eur. J. Neurosci. 20:1341–7.CrossRefPubMedGoogle Scholar
  36. 36.
    Benaroya-Milshtein N, et al. (2007) Environmental enrichment augments the efficacy of idiotype vaccination for B-cell lymphoma. J. Immunother. 30:517–22.CrossRefPubMedGoogle Scholar
  37. 37.
    Marashi V, Barnekow A, Ossendorf E, Sachser N. (2003) Effects of different forms of environmental enrichment on behavioral, endocrinological, and immunological parameters in male mice. Horm. Behav. 43:281–92.CrossRefPubMedGoogle Scholar
  38. 38.
    Marashi V, Barnekow A, Sachser N. (2004) Effects of environmental enrichment on males of a docile inbred strain of mice. Physiol. Behav. 82:765–76.CrossRefPubMedGoogle Scholar
  39. 39.
    Trullas R, Skolnick P. (1993) Differences in fear motivated behaviors among inbred mouse strains. Psychopharmacology (Berl.). 111:323–31.CrossRefGoogle Scholar
  40. 40.
    Shanks N, Griffiths J, Zalcman S, Zacharko RM, Anisman H. (1990) Mouse strain differences in plasma corticosterone following uncontrollable footshock. Pharmacol. Biochem. Behav. 36:515–9.CrossRefPubMedGoogle Scholar
  41. 41.
    Petkova SB, et al. (2008) Genetic influence on immune phenotype revealed strain-specific variations in peripheral blood lineages. Physiol. Genomics. 34:304–14.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Pruett SB, Fan R, Myers LP, Wu WJ, Collier S. (2000) Quantitative analysis of the neuroendocrine-immune axis: linear modeling of the effects of exogenous corticosterone and restraint stress on lymphocyte subpopulations in the spleen and thymus in female B6C3F1 mice. Brain. Behav. Immun. 14:270–87.CrossRefPubMedGoogle Scholar
  43. 43.
    del Rey A, Besedovsky H, Sorkin E. (1984) Endogenous blood levels of corticosterone control the immunologic cell mass and B cell activity in mice. J. Immunol. 133:572–5.PubMedGoogle Scholar
  44. 44.
    Garvy BA, King LE, Telford WG, Morford LA, Fraker PJ. (1993) Chronic elevation of plasma corticosterone causes reductions in the number of cycling cells of the B lineage in murine bone marrow and induces apoptosis. Immunology. 80:587–92.PubMedPubMedCentralGoogle Scholar
  45. 45.
    Zhong G, Reise Sousa C, Germain RN. (1997) Antigen-unspecific B cells and lymphoid dendritic cells both show extensive surface expression of processed antigen-major histocompatibility complex class II complexes after soluble protein exposure in vivo or in vitro. J. Exp. Med. 186:673–82.CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Townsend SE, Goodnow CC. (1998) Abortive proliferation of rare T cells induced by direct or indirect antigen presentation by rare B cells in vivo. J. Exp. Med. 187:1611–21.CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Kiecolt-Glaser JK, Glaser R, Gravenstein S, Malarkey WB, Sheridan J. (1996) Chronic stress alters the immune response to influenza virus vaccine in older adults. Proc. Natl. Acad. Sci. U. S. A. 93:3043–7.CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Feng N, et al. (1991) The effect of restraint stress on the kinetics, magnitude, and isotype of the humoral immune response to influenza virus infection. Brain Behav. Immun. 5:370–82.CrossRefPubMedGoogle Scholar
  49. 49.
    Karp JD, Moynihan JA, Ader R. (1993) Effects of differential housing on the primary and secondary antibody responses of male C57BL/6 and BALB/c mice. Brain Behav. Immun. 7:326–33.CrossRefPubMedGoogle Scholar
  50. 50.
    Meewisse ML, Reitsma JB, de Vries GJ, Gersons BP, Olff M. (2007) Cortisol and post-traumatic stress disorder in adults: systematic review and meta-analysis. Br. J. Psychiatry. 191:387–92.CrossRefPubMedGoogle Scholar
  51. 51.
    Daskalakis NP, Yehuda R, Diamond DM. (2013) Animal models in translational studies of PTSD. Psychoneuroendocrinology. 38:1895–911.CrossRefPubMedGoogle Scholar

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Authors and Affiliations

  • Blake T. Gurfein
    • 1
    • 2
    • 8
  • Olga Davidenko
    • 3
    • 4
  • Mary Premenko-Lanier
    • 2
  • Jeffrey M. Milush
    • 2
  • Michael Acree
    • 1
  • Mary F. Dallman
    • 5
  • Chadi Touma
    • 6
  • Rupert Palme
    • 7
  • Vanessa A. York
    • 2
  • Gilles Fromentin
    • 4
  • Nicolas Darcel
    • 3
    • 4
  • Douglas F. Nixon
    • 2
  • Frederick M. Hecht
    • 1
  1. 1.Osher Center for Integrative MedicineUniversity of CaliforniaSan FranciscoUSA
  2. 2.Division of Experimental MedicineUniversity of CaliforniaSan FranciscoUSA
  3. 3.Chaire ANCA, Food Science, Nutrition and Eating BehaviorAgroParisTechParisFrance
  4. 4.INRA, Unit 914 Nutrition Physiology and Ingestive BehaviorAgroParisTechParisFrance
  5. 5.Department of PhysiologyUniversity of CaliforniaSan FranciscoUSA
  6. 6.Research Group of PsychoneuroendocrinologyMax Planck Institute of PsychiatryMunichGermany
  7. 7.Department of Biomedical Sciences/BiochemistryUniversity of Veterinary MedicineViennaAustria
  8. 8.UCSF Division of Experimental MedicineSan FranciscoUSA

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