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IL-35 is a Protective Immunomodulator in Brain Ischemic Injury in Mice

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Abstract

IL-35 has been identified as a novel anti-inflammatory cytokine that belongs to the IL-12 cytokine family and has been verified to play a protective role in autoimmune diseases. In this study, we investigated the protective effects of IL-35 on cerebral ischemia/reperfusion (I/R) injury in a middle cerebral artery occlusion mouse model. We determined that the expression of IL-35 was initially decreased and subsequently increased in I/R injury. Moreover, IL-35 (i.c.v.) pre- and posttreatment significantly reduced the infarct volume and improved neurological deficits after 45 min of ischemia and 24 h of reperfusion. Importantly, IL-35 treatment improved neurological function recovery, particularly in balance ability, at 14 days after treatment. Finally, our results showed that IL-35 treatment reduced the expression of IL-6 and IL-1β, which are confirmed proinflammatory cytokines, thus indicating that these cytokines have both been linked to the anti-inflammatory mechanisms of IL-35. Therefore, IL-35 may be a key immune mediator in brain ischemic injury and appears to have promising potential for clinical trials.

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Abbreviations

IL:

Interleukin

CSF:

Cerebrospinal fluid

STAT:

Signal transducers and activators of transcription

KO:

Knockout

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

PBS:

Phosphate buffered solution

DMSO:

Dimethyl sulfoxide

References

  1. Sacco RL, Roth GA, Reddy KS, Arnett DK, Bonita R, Gaziano TA, Heidenreich PA, Huffman MD, Mayosi BM, Mendis S, Murray CJ, Perel P, Pineiro DJ, Smith SC Jr, Taubert KA, Wood DA, Zhao D, Zoghbi WA (2016) The Heart of 25 by 25: achieving the goal of reducing global and regional premature deaths from cardiovascular diseases and stroke: a modeling study from the American Heart Association and world heart federation. Glob Heart 11:251–264

    Article  PubMed  Google Scholar 

  2. Chen C, Chu SF, Liu DD, Zhang Z, Kong LL, Zhou X, Chen NH (2017) Chemokines play complex roles in cerebral ischemia. Neurochem Int 112:146–158

    Google Scholar 

  3. Sadatomo A, Inoue Y, Ito H, Karasawa T, Kimura H, Watanabe S, Mizushina Y, Nakamura J, Kamata R, Kasahara T, Horie H, Sata N, Takahashi M (2017) Interaction of neutrophils with macrophages promotes IL-1beta maturation and contributes to hepatic ischemia-reperfusion injury. J Immunol 199:3306–3315

    Article  PubMed  CAS  Google Scholar 

  4. Legido A, Katsetos CD (2014) Experimental studies in epilepsy: immunologic and inflammatory mechanisms. Semin Pediatr Neurol 21:197–206

    Article  PubMed  Google Scholar 

  5. Chamorro A, Urra X, Planas AM (2007) Infection after acute ischemic stroke: a manifestation of brain-induced immunodepression. Stroke 38:1097–1103

    Article  PubMed  Google Scholar 

  6. Niedbala W, Wei XQ, Cai B, Hueber AJ, Leung BP, McInnes IB, Liew FY (2007) IL-35 is a novel cytokine with therapeutic effects against collagen-induced arthritis through the expansion of regulatory T cells and suppression of Th17 cells. Eur J Immunol 37:3021–3029

    Article  PubMed  Google Scholar 

  7. Collison LW, Workman CJ, Kuo TT, Boyd K, Wang Y, Vignali KM, Cross R, Sehy D, Blumberg RS, Vignali DAA (2007) The inhibitory cytokine IL-35 contributes to regulatory T-cell function. Nature 450:566

    Article  PubMed  CAS  Google Scholar 

  8. Shen P, Roch T, Lampropoulou V, O’Connor RA, Stervbo U, Hilgenberg E, Ries S, Dang VD, Jaimes Y, Daridon C, Li R, Jouneau L, Boudinot P, Wilantri S, Sakwa I, Miyazaki Y, Leech MD, McPherson RC, Wirtz S, Neurath M, Hoehlig K, Meinl E, Grutzkau A, Grun JR, Horn K, Kuhl AA, Dorner T, Bar-Or A, Kaufmann SHE, Anderton SM, Fillatreau S (2014) IL-35-producing B cells are critical regulators of immunity during autoimmune and infectious diseases. Nature 507:366

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  9. Takeda A, Hasegawa E, Fukuhara T, Hirakawa S, Yamada H, Yang Y, Yoshimura T, Hisatomi T, Oshima Y, Yoshida H, Sonoda KH, Ishibashi T (2014) EBI3 is pivotal for the initiation of experimental autoimmune uveitis. Exp Eye Res 125:107–113

    Article  PubMed  CAS  Google Scholar 

  10. Jafarzadeh A, Jamali M, Mahdavi R, Ebrahimi HA, Hajghani H, Khosravimashizi A, Nemati M, Najafipour H, Sheikhi A, Mohammadi MM, Daneshvar H (2015) Circulating levels of interleukin-35 in patients with multiple sclerosis: evaluation of the influences of FOXP3 gene polymorphism and treatment program. J Mol Neurosci 55:891–897

    Article  PubMed  CAS  Google Scholar 

  11. Collison LW, Pillai MR, Chaturvedi V, Vignali DA (2009) Regulatory T cell suppression is potentiated by target T cells in a cell contact, IL-35- and IL-10-dependent manner. J Immunol 182:6121–6128

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  12. Zeng JC, Zhang Z, Li TY, Liang YF, Wang HM, Bao JJ, Zhang JA, Wang WD, Xiang WY, Kong B, Wang ZY, Wu BH, Chen XD, He L, Zhang S, Wang CY, Xu JF (2013) Assessing the role of IL-35 in colorectal cancer progression and prognosis. Int J Clin Exp Pathol 6:1806–1816

    PubMed  PubMed Central  Google Scholar 

  13. Manzoor F, Johnson MC, Li C, Samulski RJ, Wang B, Tisch R (2017) Beta-cell-specific IL-35 therapy suppresses ongoing autoimmune diabetes in NOD mice. Eur J Immunol 47:144–154

    Article  PubMed  CAS  Google Scholar 

  14. Wu H, Li P, Shao N, Ma J, Ji M, Sun X, Ma D, Ji C (2012) Aberrant expression of Treg-associated cytokine IL-35 along with IL-10 and TGF-beta in acute myeloid leukemia. Oncol Lett 3:1119–1123

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Frieler RA, Meng H, Duan SZ, Berger S, Schutz G, He Y, Xi G, Wang MM, Mortensen RM (2011) Myeloid-specific deletion of the mineralocorticoid receptor reduces infarct volume and alters inflammation during cerebral ischemia. Stroke 42:179–185

    Article  PubMed  Google Scholar 

  16. Prieto-Moure B, Lloris-Carsi JM, Belda-Antoli M, Toledo-Pereyra LH, Cejalvo-Lapena D (2017) Allopurinol protective effect of renal ischemia by downregulating TNF-alpha, IL-1beta, and IL-6 response. J Invest Surg 30:143–151

    Article  PubMed  Google Scholar 

  17. Ohtaki H, Nakamachi T, Dohi K, Aizawa Y, Takaki A, Hodoyama K, Yofu S, Hashimoto H, Shintani N, Baba A, Kopf M, Iwakura Y, Matsuda K, Arimura A, Shioda S (2006) Pituitary adenylate cyclase-activating polypeptide (PACAP) decreases ischemic neuronal cell death in association with IL-6. Proc Natl Acad Sci USA 103:7488–7493

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  18. Tarkowski E, Rosengren L, Blomstrand C, Wikkelso C, Jensen C, Ekholm S, Tarkowski A (1995) Early intrathecal production of interleukin-6 predicts the size of brain lesion in stroke. Stroke 26:1393–1398

    Article  PubMed  CAS  Google Scholar 

  19. Collison LW, Delgoffe GM, Guy CS, Vignali KM, Chaturvedi V, Fairweather D, Satoskar AR, Garcia KC, Hunter CA, Drake CG, Murray PJ, Vignali DAA (2012) The composition and signaling of the IL-35 receptor are unconventional. Nat Immunol 13:290

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  20. Xu X, Chua CC, Gao J, Hamdy RC, Chua BH (2006) Humanin is a novel neuroprotective agent against stroke. Stroke 37:2613–2619

    Article  PubMed  CAS  Google Scholar 

  21. Cavalla D, Hadjiconstantinou M, Laird HE 2nd, Neff NH (1985) Intracerebroventricular administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and its metabolite 1-methyl-4-phenylpyridinium ion (MPP+) decrease dopamine and increase acetylcholine in the mouse neostriatum. Neuropharmacology 24:585–586

    Article  PubMed  CAS  Google Scholar 

  22. Xu X, Chua CC, Gao J, Chua KW, Wang H, Hamdy RC, Chua BH (2008) Neuroprotective effect of humanin on cerebral ischemia/reperfusion injury is mediated by a PI3K/Akt pathway. Brain Res 1227:12–18

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Yen TL, Chen RJ, Jayakumar T, Lu WJ, Hsieh CY, Hsu MJ, Yang CH, Chang CC, Lin YK, Lin KH, Sheu JR (2016) Andrographolide stimulates p38 mitogen-activated protein kinase-nuclear factor erythroid-2-related factor 2-heme oxygenase 1 signaling in primary cerebral endothelial cells for definite protection against ischemic stroke in rats. Transl Res 170:57–72

    Article  PubMed  CAS  Google Scholar 

  24. Jiang X, Pu H, Hu X, Wei Z, Hong D, Zhang W, Gao Y, Chen J, Shi Y (2016) A post-stroke therapeutic regimen with omega-3 polyunsaturated fatty acids that promotes white matter integrity and beneficial microglial responses after cerebral ischemia. Transl Stroke Res 7:548–561

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Schallert T, Fleming SM, Leasure JL, Tillerson JL, Bland ST (2000) CNS plasticity and assessment of forelimb sensorimotor outcome in unilateral rat models of stroke, cortical ablation, parkinsonism and spinal cord injury. Neuropharmacology 39:777–787

    Article  PubMed  CAS  Google Scholar 

  26. Lin Y, Huang Y, Lu Z, Luo C, shi Y, Zeng Q, Cao Y, Liu L, Wang X, Ji Q (2012) Decreased plasma IL-35 levels are related to the left ventricular ejection fraction in coronary artery diseases. PLoS ONE 7:e52490

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  27. Treib J, Grauer MT, Woessner R, Morgenthaler M (2000) Treatment of stroke on an intensive stroke unit: a novel concept. Intensive Care Med 26:1598–1611

    Article  PubMed  CAS  Google Scholar 

  28. Di Napoli M, Papa F (2003) The proteasome system and proteasome inhibitors in stroke: controlling the inflammatory response. Curr Opin Investig Drugs 4:1333–1342

    PubMed  Google Scholar 

  29. Irmler IM, Gajda M, Kamradt T (2014) Amelioration of experimental arthritis by stroke-induced immunosuppression is independent of Treg cell function. Ann Rheum Dis 73:2183–2191

    Article  PubMed  CAS  Google Scholar 

  30. Li P, Mao L, Liu X, Gan Y, Zheng J, Thomson AW, Gao Y, Chen J, Hu X (2014) Essential role of program death 1-ligand 1 in regulatory T-cell-afforded protection against blood-brain barrier damage after stroke. Stroke 45:857–864

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  31. Yang XF, Li XY, Mai JT, Virtue A, Yin Y, Sha XJ, Wang H (2012) IL-35 is a novel responsive anti-inflammatory cytokine—a new system of categorizing anti-inflammatory cytokines. Blood 120:e33628

    Google Scholar 

  32. Kochetkova I, Golden S, Callis G, Pascual DW (2009) Anti-inflammatory properties of IL-35 promote induction of regulatory CD4(+)T cells to suppress collagen II-induced arthritis (CIA). J Immunol 182:99

    Google Scholar 

  33. Chen GH, Liang YF, Guan X, Chen H, Liu QK, Lin BH, Chen C, Huang MY, Chen JA, Wu WQ, Liang Y, Zhou KY, Zeng JC (2016) Circulating low IL-23: IL-35 cytokine ratio promotes progression associated with poor prognosisin breast cancer. Am J Transl Res 8:2255

    PubMed  PubMed Central  CAS  Google Scholar 

  34. Yang GY, Schielke GP, Gong C, Mao Y, Ge HL, Liu XH, Betz AL (1999) Expression of tumor necrosis factor-alpha and intercellular adhesion molecule-1 after focal cerebral ischemia in interleukin-1beta converting enzyme deficient mice. J Cereb Blood Flow Metab 19:1109–1117

    Article  PubMed  CAS  Google Scholar 

  35. Perini F, Morra M, Alecci M, Galloni E, Marchi M, Toso V (2001) Temporal profile of serum anti-inflammatory and pro-inflammatory interleukins in acute ischemic stroke patients. Neurol Sci 22:289–296

    Article  PubMed  CAS  Google Scholar 

  36. Chaturvedi V, Collison LW, Guy CS, Workman CJ, Vignali DA (2011) Cutting edge: human regulatory T cells require IL-35 to mediate suppression and infectious tolerance. J Immunol 186:6661–6666

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  37. Collison LW, Delgoffe GM, Guy CS, Vignali KM, Chaturvedi V, Fairweather D, Satoskar AR, Garcia KC, Hunter CA, Drake CG, Murray PJ, Vignali DA (2012) The composition and signaling of the IL-35 receptor are unconventional. Nat Immunol 13:290–299

    Article  PubMed  PubMed Central  CAS  Google Scholar 

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Acknowledgements

This work was supported by the Neurology Clinical Research Foundation of Suzhou. The funders have no role in the study design or data collection.

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  1. Chen Xu and Hao Zhu are co-first authors.

Authors and Affiliations

  1. Department of Neurology, The Affiliated Suzhou Hospital, Nanjing Medical University, Suzhou, China

    Chen Xu, Hao Zhu, Rong Shen, Qian Feng, Hua Zhou & Zhong Zhao

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  1. Chen Xu
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  2. Hao Zhu
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  5. Hua Zhou
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Contributions

All authors were involved in drafting the article or critically revising it for important intellectual content, and all authors approved the final version for publication. ZZ and ZH conceived and supervised the project; XC and ZHao wrote the manuscript and generated the final figures. All experimental procedures and data analysis were performed by XC, ZH, SR and FQ; XC and FQ contributed to test behavioral experiments; XC and ZHao contributed to data analysis.

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Correspondence to Hua Zhou or Zhong Zhao.

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Xu, C., Zhu, H., Shen, R. et al. IL-35 is a Protective Immunomodulator in Brain Ischemic Injury in Mice. Neurochem Res 43, 1454–1463 (2018). https://doi.org/10.1007/s11064-018-2560-5

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  • DOI: https://doi.org/10.1007/s11064-018-2560-5

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