Metabolic Brain Disease

, Volume 33, Issue 5, pp 1599–1607 | Cite as

Antibiotics protect against EAE by increasing regulatory and anti-inflammatory cells

  • Hilary A. Seifert
  • Gil Benedek
  • Ha Nguyen
  • Grant Gerstner
  • Ying Zhang
  • Gail Kent
  • Arthur A. Vandenbark
  • Jürgen Bernhagen
  • Halina Offner
Original Article


A seven day pretreatment course of an oral antibiotic cocktail (Ampicillin, Metronidazole, Neomycin Sulfate, and Vancomycin) was shown to induce changes in peripheral immune regulation and protect mice from signs of experimental autoimmune encephalomyelitis (EAE). To determine if a shorter course of antibiotic pretreatment could also protect the mice from EAE and induce regulatory immune cells, studies were conducted using the same oral antibiotic cocktail for three days. In addition, the CNS was examined to determine the effects of antibiotic pretreatment on EAE disease course and immune modulation within the affected tissue. The shorter three day pretreatment course was also significantly protective against severe EAE in C57BL/6 mice. Moreover, our study found increased frequencies of regulatory cells and a decrease in the frequency of anti-inflammatory macrophages in the spleen of EAE protected mice. Additionally, a chemokine and chemokine receptor array run on mRNA from spinal cords revealed that genes associated with regulatory T cells and macrophage recruitment were strongly upregulated in the antibiotic pretreated mice. Additional RT-PCR data showed genes associated with anti-inflammatory microglia/macrophages were upregulated and pro-inflammatory genes were downregulated. This suggests the macrophages recruited to the spinal cord by chemokines are subsequently polarized toward an anti-inflammatory phenotype. These results lend strong support to the conclusion that a three day course of antibiotic treatment given prior to the induction of severe EAE profoundly protected the mice by inducing regulatory lymphocytes in the periphery and an anti-inflammatory milieu in the affected spinal cord tissue.


Microbiota Neuroinflammation EAE Antibiotic Regulatory cells CNS 



This work was supported by Deutsche Forschungsgemeinschaft (DFG) EXC1010 SyNergy (JB), NIH, National Institute Of Neurological Disorders And Stroke of the National Institutes R01NS080890 (HO) and the Department of Veterans Affairs IO1 BX000226-09 and 1 IK6 BX004209 (AAV). This material is also the result of work supported with resources and the use of facilities at the VA Portland Health Care Center in Portland, Oregon. The contents do not represent the views of the U.S. Department of Veterans Affairs, the United States Government or the official views of the National Institutes of Health.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human and/or animals

All applicable international, national and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. This article does not contain any studies with human participants performed by any of the authors.


  1. Berer K et al (2011) Commensal microbiota and myelin autoantigen cooperate to trigger autoimmune demyelination. Nature 479:538–541. CrossRefPubMedGoogle Scholar
  2. Bodhankar S, Chen Y, Lapato A, Vandenbark AA, Murphy SJ, Saugstad JA, Offner H (2015) Regulatory CD8(+)CD122 (+) T-cells predominate in CNS after treatment of experimental stroke in male mice with IL-10-secreting B-cells. Metab Brain Dis 30:911–924. CrossRefPubMedGoogle Scholar
  3. Dziembowska M, Tham TN, Lau P, Vitry S, Lazarini F, Dubois-Dalcq M (2005) A role for CXCR4 signaling in survival and migration of neural and oligodendrocyte precursors. Glia 50:258–269. CrossRefPubMedGoogle Scholar
  4. Erny D et al (2015) Host microbiota constantly control maturation and function of microglia in the CNS. Nat Neurosci 18:965–977. CrossRefPubMedPubMedCentralGoogle Scholar
  5. Fararjeh M, Mohammad MK, Bustanji Y, Alkhatib H, Abdalla S (2008) Evaluation of immunosuppression induced by metronidazole in Balb/c mice and human peripheral blood lymphocytes. Int Immunopharmacol 8:341–350. CrossRefPubMedGoogle Scholar
  6. Freedman SN, Shahi SK, Mangalam AK (2018) The "gut feeling": breaking down the role of gut microbiome in multiple sclerosis. Neurotherapeutics 15:109–125. CrossRefPubMedGoogle Scholar
  7. Ivanov II et al (2008) Specific microbiota direct the differentiation of IL-17-producing T-helper cells in the mucosa of the small intestine. Cell Host Microbe 4:337–349. CrossRefPubMedPubMedCentralGoogle Scholar
  8. Iwata Y et al (2011) Characterization of a rare IL-10-competent B-cell subset in humans that parallels mouse regulatory B10 cells. Blood 117:530–541. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Kowarik MC et al (2012) CXCL13 is the major determinant for B cell recruitment to the CSF during neuroinflammation. J Neuroinflammation 9:93. CrossRefPubMedPubMedCentralGoogle Scholar
  10. Lee YK, Menezes JS, Umesaki Y, Mazmanian SK (2011) Proinflammatory T-cell responses to gut microbiota promote experimental autoimmune encephalomyelitis. Proc Natl Acad Sci U S A 108(Suppl 1):4615–4622. CrossRefPubMedGoogle Scholar
  11. Lim HW, Lee J, Hillsamer P, Kim CH (2008) Human Th17 cells share major trafficking receptors with both polarized effector T cells and FOXP3+ regulatory T cells. J Immunol 180:122–129CrossRefPubMedGoogle Scholar
  12. Liu J et al (2008) IFN-gamma and IL-17 production in experimental autoimmune encephalomyelitis depends on local APC-T cell complement production. J Immunol 180:5882–5889CrossRefPubMedPubMedCentralGoogle Scholar
  13. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods 25:402–408. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Matsumoto M et al (2014) Interleukin-10-producing plasmablasts exert regulatory function in autoimmune inflammation. Immunity 41:1040–1051. CrossRefPubMedGoogle Scholar
  15. Meiron M, Zohar Y, Anunu R, Wildbaum G, Karin N (2008) CXCL12 (SDF-1alpha) suppresses ongoing experimental autoimmune encephalomyelitis by selecting antigen-specific regulatory T cells. J Exp Med 205:2643–2655. CrossRefPubMedPubMedCentralGoogle Scholar
  16. Mielcarz DW, Kasper LH (2015) The gut microbiome in multiple sclerosis. Curr Treat Options Neurol 17:344. CrossRefPubMedGoogle Scholar
  17. Niess JH, Leithauser F, Adler G, Reimann J (2008) Commensal gut flora drives the expansion of proinflammatory CD4 T cells in the colonic lamina propria under normal and inflammatory conditions. J Immunol 180:559–568CrossRefPubMedGoogle Scholar
  18. Ochoa-Reparaz J, Kasper LH (2014) Gut microbiome and the risk factors in central nervous system autoimmunity. FEBS Lett 588:4214–4222. CrossRefPubMedPubMedCentralGoogle Scholar
  19. Ochoa-Reparaz J, Mielcarz DW, Ditrio LE, Burroughs AR, Foureau DM, Haque-Begum S, Kasper LH (2009) Role of gut commensal microflora in the development of experimental autoimmune encephalomyelitis. J Immunol 183:6041–6050. CrossRefPubMedGoogle Scholar
  20. Ochoa-Reparaz J, Mielcarz DW, Haque-Begum S, Kasper LH (2010) Induction of a regulatory B cell population in experimental allergic encephalomyelitis by alteration of the gut commensal microflora. Gut Microbes 1:103–108. CrossRefPubMedPubMedCentralGoogle Scholar
  21. Palm NW, de Zoete MR, Flavell RA (2015) Immune-microbiota interactions in health and disease. Clin Immunol 159:122–127. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Rupprecht TA et al (2009) The chemokine CXCL13 is a key regulator of B cell recruitment to the cerebrospinal fluid in acute Lyme neuroborreliosis. J Neuroinflammation 6:42. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Shi Z, Rifa'i M, Lee YH, Shiku H, Isobe K, Suzuki H (2008) Importance of CD80/CD86-CD28 interactions in the recognition of target cells by CD8+CD122+ regulatory T cells. Immunology 124:121–128. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Wang Y et al (2014) A commensal bacterial product elicits and modulates migratory capacity of CD39(+) CD4 T regulatory subsets in the suppression of neuroinflammation. Gut Microbes 5:552–561. CrossRefPubMedGoogle Scholar
  25. Zhang J, Lapato A, Bodhankar S, Vandenbark AA, Offner H (2015) Treatment with IL-10 producing B cells in combination with E2 ameliorates EAE severity and decreases CNS inflammation in B cell-deficient mice. Metab Brain Dis 30:1117–1127. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of NeurologyOregon Health & Science UniversityPortlandUSA
  2. 2.Neuroimmunology ResearchVeterans Affairs Portland Health Care System, R&D-31PortlandUSA
  3. 3.Tissue Typing and Immunogenetics Laboratory, Hadassah Medical CenterJerusalemIsrael
  4. 4.Department of Molecular Microbiology & ImmunologyOregon Health & Science UniversityPortlandUSA
  5. 5.Vascular Biology, Institute for Stroke and Dementia ResearchKlinikum der Universität München, Ludwig-Maximilians-University MunichMunichGermany
  6. 6.Munich Cluster for Systems Neurology (EXC 1010 SyNergy)MunichGermany
  7. 7.Department of Anesthesiology & Perioperative MedicineOregon Health & Science UniversityPortlandUSA

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