Abstract
We investigated changes in innate and adaptive immunity paralleling the progressive nigrostriatal damage occurring in a neurotoxic model of Parkinson’s disease (PD) based on unilateral infusion of 6-hydroxydopamine (6-OHDA) into the rat striatum. A time-course analysis was conducted to assess changes in morphology (activation) and cell density of microglia and astrocytes, microglia polarization (M1 vs. M2 phenotype), lymphocyte infiltration in the lesioned substantia nigra pars compacta (SNc), and modifications of CD8+ and subsets of CD4+ T cell in peripheral blood accompanying nigrostriatal degeneration. Confirming previous results, we observed slightly different profiles of activation for astrocytes and microglia paralleling nigral neuronal loss. For astrocytes, morphological changes and cell density increases were mostly evident at the latest time points (14 and 28 days post-surgery), while moderate microglia activation was present since the earliest time point. For the first time, in this model, we described the time-dependent profile of microglia polarization. Activated microglia clearly expressed the M2 phenotype in the earlier phase of the experiment, before cell death became manifest, gradually shifting to the M1 phenotype as SNc cell death started. In parallel, a reduction in the percentage of circulating CD4+ T regulatory (Treg) cells, starting as early as day 3 post-6-OHDA injection, was detected in 6-OHDA-injected rats. Our data show that nigrostriatal degeneration is associated with complex changes in central and peripheral immunity. Microglia activation and polarization, Treg cells, and the factors involved in their cross-talk should be further investigated as targets for the development of therapeutic strategies for disease modification in PD.
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References
Ambrosi G, Armentero MT, Levandis G, Bramanti P, Nappi GBF (2010) Effects of early and delayed treatment with an mGluR5 antagonist on motor impairment, nigrostriatal damage and neuroinflammation in a rodent model of Parkinson’s disease. Brain Res Bull 82:29–38. doi:10.1016/j.brainresbull.2010.01.011
Ambrosi G, Ghezzi C, Sepe S, Milanese C, Payan-Gomez C, Bombardieri CR, Armentero MT, Zangaglia R, Pacchetti C, Mastroberardino PGBF (2014) Bioenergetic and proteolytic defects in fibroblasts from patients with sporadic Parkinson’s disease. Biochim Biophys Acta 1842:1385–1394. doi:10.1016/j.bbadis.2014.05.008
Appel SH (2009) CD4+ T cells mediate cytotoxicity in neurodegenerative diseases. J Clin Invest 119:13–15. doi:10.1172/JCI38096
Armentero MT, Levandis G, Nappi G, Bazzini EBF (2006) Peripheral inflammation and neuroprotection: systemic pretreatment with complete Freund’s adjuvant reduces 6-hydroxydopamine toxicity in a rodent model of Parkinson's disease. Neurobiol Dis 24:492–505. doi:10.1016/j.nbd.2006.08.016
Armentero MT, Levandis G, Bazzini E, Cerri S, Ghezzi CBF (2011) Adhesion molecules as potential targets for neuroprotection in a rodent model of Parkinson’s disease. Neurobiol Dis 43:663–668. doi:10.1016/j.nbd.2011.05.017
Baba Y, Kuroiwa A, Uitti RJ, Wszolek ZKYT (2005) Alterations of T-lymphocyte populations in Parkinson disease. Park Relat Disord 11:493–498. doi:10.1016/j.parkreldis.2005.07.005
Barcia C, Ros CM, Annese V et al (2011) IFN-γ signaling, with the synergistic contribution of TNF-α, mediates cell specific microglial and astroglial activation in experimental models of Parkinson’s disease. Cell Death Dis 2:e142. doi:10.1038/cddis.2011.17
Blandini F (2013) Neural and immune mechanisms in the pathogenesis of Parkinson’s disease. J NeuroImmune Pharmacol 8:189–201. doi:10.1007/s11481-013-9435-y
Blandini F, Levandis G, Bazzini E et al (2007) Time-course of nigrostriatal damage, basal ganglia metabolic changes and behavioural alterations following intrastriatal injection of 6-hydroxydopamine in the rat: new clues from an old model. Eur J Neurosci 25:397–405. doi:10.1111/j.1460-9568.2006.05285.x
Brochard V, Combadière B, Prigent A et al (2009) Infiltration of CD4+ lymphocytes into the brain contributes to neurodegeneration in a mouse model of Parkinson disease. J Clin Invest 119:182–192. doi:10.1172/JCI36470
Bruchelt G, Grygar G, Treuner J, Esterbauer H, Niethammer D (1989) Cytotoxic effects of 6-hydroxydopamine, merocyanine-540 and related compounds on human neuroblastoma and hematopoietic stem cells. Free Radic Res Commun 7:205–212
Carvey PM, Zhao CH, Hendey B et al (2005) 6-Hydroxydopamine-induced alterations in blood-brain barrier permeability. Eur J Neurosci 22:1158–1168. doi:10.1111/j.1460-9568.2005.04281.x
Chaturvedi RK, Beal M (2013) Mitochondria targeted therapeutic approaches in Parkinson’s and Huntington's diseases. Mol Cell Neurosci 55:101–114. doi:10.1016/j.mcn.2012.11.011
Chaudhuri KR, Odin P, Antonini A-MP (2011) Parkinson’s disease: the non-motor issues. Park Relat Disord 17:717–723
Colburn RW, DeLeo JA, Rickman AJ, Yeager MP, Kwon PHW (1997) Dissociation of microglial activation and neuropathic pain behaviors following peripheral nerve injury in the rat. J Neuroimmunol 79:163–175
Doorn KJ, Lucassen PJ, Boddeke HW, Prins M, Berendse HW, Drukarch B, van Dam AM (2012) Emerging roles of microglial activation and non-motor symptoms in Parkinson’s disease. Prog Neurobiol 98:222–238. doi:10.1016/j.pneurobio.2012.06.005
Double KL, Reyes S, Werry EL HG (2010) Selective cell death in neurodegeneration: why are some neurons spared in vulnerable regions? Progr Neurobiol 92:316–329. doi:10.1016/j.pneurobio.2010.06.001
Espinosa-Oliva AM, de Pablos RM, Sarmiento M, Villarán RF, Carrillo-Jiménez A, Santiago M, Venero JL, Herrera AJ, Cano JMA (2014) Role of dopamine in the recruitment of immune cells to the nigro-striatal dopaminergic structures. Neurotoxicology 41:89–101. doi:10.1016/j.neuro.2014.01.006
González H, Pacheco R (2014) T-cell-mediated regulation of neuroinflammation involved in neurodegenerative diseases. J Neuroinflammation 11:201. doi:10.1186/s12974-014-0201-8
González-Hernández T, Cruz-Muros I, Afonso-Oramas D et al (2010) Vulnerability of mesostriatal dopaminergic neurons in Parkinson’s disease. Front Neuroanat 4:140. doi:10.3389/fnana.2010.00140
Greenamyre JT, Hastings TG (2004) Biomedicine. Parkinson’s—divergent causes, convergent mechanisms. Science 304:1120–1122. doi:10.1126/science.1098966
Hauser DN, Hastings TG (2013) Mitochondrial dysfunction and oxidative stress in Parkinson’s disease and monogenic parkinsonism. Neurobiol Dis 51:35–42. doi:10.1016/j.nbd.2012.10.011
He F, Balling R (2013) The role of regulatory T cells in neurodegenerative diseases. Wiley Interdiscip Rev Syst Biol Med 5:153–180. doi:10.1002/wsbm.1187
Herrero M-T, Estrada C, Maatouk L, Vyas S (2015) Inflammation in Parkinson’s disease: role of glucocorticoids. Front Neuroanat 9:32. doi:10.3389/fnana.2015.00032
Hirsch EC, Hunot S (2009) Neuroinflammation in Parkinson's disease: a target for neuroprotection? Lancet Neurol 8:382–397
Hirsch EC, Vyas SHS (2012) Neuroinflammation in Parkinson’s disease. Park Relat Disord 18:S210–S212. doi:10.1016/S1353-8020(11)70065-7
Hu X, Li P, Guo Y et al (2012) Microglia/macrophage polarization dynamics reveal novel mechanism of injury expansion after focal cerebral ischemia. Stroke 43:3063–3070. doi:10.1161/STROKEAHA.112.659656
Kannarkat GT, Boss JM, Tansey MG (2013) The role of innate and adaptive immunity in Parkinson’s disease. J Parkinsons Dis 3:493–514. doi:10.3233/JPD-130250
Kipnis J, Cardon M, Avidan H et al (2004) Dopamine, through the extracellular signal-regulated kinase pathway, downregulates CD4+CD25+ regulatory T-cell activity: implications for neurodegeneration. J Neurosci 24:6133–6143. doi:10.1523/JNEUROSCI.0600-04.2004
Kitamura Y, Inden M, Minamino H, Abe M, Takata K, Taniguchi T (2010) The 6-hydroxydopamine-induced nigrostriatal neurodegeneration produces microglia-like NG2 glial cells in the rat substantia nigra. Glia 58(14):1686–1700. doi:10.1002/glia.21040
Kustrimovic N, Rasini E, Legnaro M et al (2014) Expression of dopaminergic receptors on human CD4+ T lymphocytes: flow cytometric analysis of naive and memory subsets and relevance for the neuroimmunology of neurodegenerative disease. J NeuroImmune Pharmacol 9:302–312. doi:10.1007/s11481-014-9541-5
Kustrimovic N, Rasini E, Legnaro M et al (2016) Dopaminergic receptors on CD4+ T naive and memory lymphocytes correlate with motor impairment in patients with Parkinson's disease. Sci Rep 6:33738. doi:10.1038/srep33738
Levite M (2016) Dopamine and T cells: dopamine receptors and potent effects on T cells, dopamine production in T cells, and abnormalities in the dopaminergic system in T cells in autoimmune, neurological and psychiatric diseases. Acta Physiol (Oxf) 216:42–89. doi:10.1111/apha.12476
Lowther DE, Hafler DA (2012) Regulatory T cells in the central nervous system. Immunol Rev 248:156–169. doi:10.1111/j.1600-065X.2012.01130.x
Lucin KM, Wyss-Coray T (2009) Immune activation in brain aging and neurodegeneration: too much or too little? Neuron 64:110–122. doi:10.1016/j.neuron.2009.08.039
Lull ME, Block ML (2010) Microglial activation and chronic neurodegeneration. Neurotherapeutics 7:354–365
Mak SK, McCormack AL, Manning-Bog AB et al (2010) Lysosomal degradation of alpha-synuclein in vivo. J Biol Chem 285:13621–13629. doi:10.1074/jbc.M109.074617
Martinez-Pasamar S, Abad E, Moreno B et al (2013) Dynamic cross-regulation of antigen-specific effector and regulatory T cell subpopulations and microglia in brain autoimmunity. BMC Syst Biol 7:34. doi:10.1186/1752-0509-7-34
Massano J, Bhatia KP (2012) Clinical approach to Parkinson’s disease: features, diagnosis, and principles of management. Cold Spring Harb Perspect Med 2:a008870. doi:10.1101/cshperspect.a008870
McGeer PL, McGeer EG (2008) Glial reactions in Parkinson’s disease. Mov Disord 23:474–483. doi:10.1002/mds.21751
McNaught KSP, Jackson T, JnoBaptiste R et al (2006) Proteasomal dysfunction in sporadic Parkinson’s disease. Neurology 66:S37–S49
Miklossy J, Doudet DD, Schwab C, Yu S, McGeer EGMP (2006) Role of ICAM-1 in persisting inflammation in Parkinson disease and MPTP monkeys. Exp Neurol 192:275–283. doi:10.1016/j.expneurol.2005.10.034
Miller KR, Streit WJ (2007) The effects of aging, injury and disease on microglial function: a case for cellular senescence. Neuron Glia Biol 3:245–253. doi:10.1017/S1740925X08000136
Perego C, Fumagalli S, De Simoni M-G (2011) Temporal pattern of expression and colocalization of microglia/macrophage phenotype markers following brain ischemic injury in mice. J Neuroinflammation 8:174. doi:10.1186/1742-2094-8-174
Ramesh G, MacLean AG, Philipp MT (2013) Cytokines and chemokines at the crossroads of neuroinflammation, neurodegeneration, and neuropathic pain. Mediat Inflamm 2013:480739. doi:10.1155/2013/480739
Reynolds AD, Banerjee R, Liu J, Gendelman HE, Mosley RL (2007) Neuroprotective activities of CD4þCD25þ regulatory T cells in an animal model of Parkinson’s disease. J Leuk Biol 82:1083–1094
Reynolds AD, Stone DK, Mosley RL, Gendelman HE (2009) Proteomic studies of nitrated alpha-synuclein microglia regulation by CD4+CD25+ T cells. J Proteome Res 8:3497–3511. doi:10.1021/pr9001614
Reynolds AD, Stone DK, Hutter JAL et al (2010) Regulatory T cells attenuate Th17 cell-mediated nigrostriatal dopaminergic neurodegeneration in a model of Parkinson’s disease. J Immunol 184:2261–2271. doi:10.4049/jimmunol.0901852
Shechter R, London A, Schwartz M (2013) Orchestrated leukocyte recruitment to immune-privileged sites: absolute barriers versus educational gates. Nat Rev Immunol 13:206–218. doi:10.1038/nri3391
Spillantini MG, Schmidt ML, Lee VM et al (1997) Alpha-synuclein in Lewy bodies. Nature 388:839–840. doi:10.1038/42166
Stephens LA, Malpass KH, Anderton SM (2009) Curing CNS autoimmune disease with myelin-reactive Foxp3+ Treg. Eur J Immunol 39:1108–1117. doi:10.1002/eji.200839073
Su X, Federoff HJ (2014) Immune responses in Parkinson’s disease: interplay between central and peripheral immune systems. Biomed Res Int 2014:275178. doi:10.1155/2014/275178
Theodore S, Maragos W (2015) 6-Hydroxydopamine as a tool to understand adaptive immune system-induced dopamine neurodegeneration in Parkinson’s disease. Immunopharmacol Immunotoxicol 37:393–399. doi:10.3109/08923973.2015.1070172
Vairetti M, Ferrigno A, Rizzo V, Ambrosi G, Bianchi A, Richelmi P, Blandini F, Armentero MT (2012) Impaired hepatic function and central dopaminergic denervation in a rodent model of Parkinson's disease: a self-perpetuating crosstalk? Biochim Biophys Acta 1822:176–184
Wheeler CJ, Seksenyan A, Koronyo Y et al (2014) T-lymphocyte deficiency exacerbates behavioral deficits in the 6-OHDA unilateral lesion rat model for Parkinson’s disease. J Neurol Neurophysiol. doi:10.4172/2155-9562.1000209
Xie L, Choudhury GR, Winters A et al (2015) Cerebral regulatory T cells restrain microglia/macrophage-mediated inflammatory responses via IL-10. Eur J Immunol 45:180–191. doi:10.1002/eji.201444823
Acknowledgements
This study was supported by a grant from Fondazione CARIPLO to Marco Cosentino and Fabio Blandini (Project 2011-0504: Dopaminergic modulation of CD4+ T lymphocytes: relevance for neurodegeneration and neuroprotection in Parkinson’s disease—the dopaminergic neuro-immune connection). Natasa Kustrimovic (postdoc fellow) and Cristina Ghezzi (lab technician) appointments were supported by the grant. We would like to acknowledge Dr. Marco Gnesi for performing the statistical analysis of data and correlations. The skillful technical assistance of Dr. Emanuela Rasini (Center for Research in Medical Pharmacology, University of Insubria) in the development and validation of flow cytometric assays as well as in data analysis is also gratefully acknowledged.
Author Contribution
FB, MC, and FM conceived and designed the study. GA, NK, FS, CG, SC, SC, GD and ER acquired data. GA, NK, FS, MC, and FB analyzed and interpreted data. All authors were involved in drafting the article or revising it critically for important intellectual content, and all authors approved the final version to be published. All authors agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved and declare to have confidence in the integrity of the contributions of their co-authors.
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Supplementary Fig. 1
Representative image of microglia polarization at different time points (24 h, 7 and 14 days post-6-OHDA injection) in the lesioned SNc of a 6-OHDA-treated rat. Blue signal: DAPI (nuclei); green signal: CD11b+ cells; red signal: CD32+/CD206+ cells, respectively, M1 and M2 phenotype. (GIF 501 kb)
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Ambrosi, G., Kustrimovic, N., Siani, F. et al. Complex Changes in the Innate and Adaptive Immunity Accompany Progressive Degeneration of the Nigrostriatal Pathway Induced by Intrastriatal Injection of 6-Hydroxydopamine in the Rat. Neurotox Res 32, 71–81 (2017). https://doi.org/10.1007/s12640-017-9712-2
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DOI: https://doi.org/10.1007/s12640-017-9712-2