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Immunohistochemical analysis of spinal cord components in mouse model of experimental autoimmune encephalomyelitis
Affiliations- Department of Histology, Jagiellonian University Medical College, Krakow, Poland
- Department of Neurology, Medical University of Lodz, Lodz, Poland
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Abstract
Introduction. Experimental autoimmune encephalomyelitis (EAE) is a widely used animal model for studying immunopathology of multiple sclerosis (MS) because it repeats the hallmarks of the human disease, such as focal inflammation and demyelination of the central nervous system, subsequently leading to axonal and neuronal loss. The interrelationships, timing and sequence of different pathological processes that lead to histologically observed lesions in SM are still incompletely understood.
Material and methods. EAE was induced in female C57Bl/6 mice by active immunization with MOG35-55 antigen. Development of the neurological symptoms in the animals was monitored and on that basis spinal cords were collected in three successive phases of the disease (onset, peak, chronic). Total leukocytes, T cells, macrophages/microglia, oligodendrocytes, damaged axons and surviving neuronal cell bodies were visualized using appropriate immunohistochemical markers and their density was quantitatively assessed using image analysis software.
Results. The density of all studied cells except neurons was significantly higher in EAE mice than in the control mice. The density of total leukocytes, T cells, and damaged axons increased from the onset to the peak phase and decreased in the chronic phase to reach values lower than those in the peak phase. The density of macrophages/microglia increased in the peak phase and remained at the elevated level in the chronic phase. Oligodendrocytes showed the highest density in the onset phase and gradually decreased afterwards. The density of neuronal cell bodies decreased only in the chronic phase of the disease.
Conclusions. In mouse model of EAE, inflammatory cells predominate in the early phases of the disease. This study shows for the first time that inflammation precedes oligodendrocyte death and neuronal loss and that macrophages/ microglia are the only cells persisting in large numbers in the chronic phase of the disease, probably because of the switch from proinflammatory to anti-inflammatory phenotype.
Abstract
Introduction. Experimental autoimmune encephalomyelitis (EAE) is a widely used animal model for studying immunopathology of multiple sclerosis (MS) because it repeats the hallmarks of the human disease, such as focal inflammation and demyelination of the central nervous system, subsequently leading to axonal and neuronal loss. The interrelationships, timing and sequence of different pathological processes that lead to histologically observed lesions in SM are still incompletely understood.
Material and methods. EAE was induced in female C57Bl/6 mice by active immunization with MOG35-55 antigen. Development of the neurological symptoms in the animals was monitored and on that basis spinal cords were collected in three successive phases of the disease (onset, peak, chronic). Total leukocytes, T cells, macrophages/microglia, oligodendrocytes, damaged axons and surviving neuronal cell bodies were visualized using appropriate immunohistochemical markers and their density was quantitatively assessed using image analysis software.
Results. The density of all studied cells except neurons was significantly higher in EAE mice than in the control mice. The density of total leukocytes, T cells, and damaged axons increased from the onset to the peak phase and decreased in the chronic phase to reach values lower than those in the peak phase. The density of macrophages/microglia increased in the peak phase and remained at the elevated level in the chronic phase. Oligodendrocytes showed the highest density in the onset phase and gradually decreased afterwards. The density of neuronal cell bodies decreased only in the chronic phase of the disease.
Conclusions. In mouse model of EAE, inflammatory cells predominate in the early phases of the disease. This study shows for the first time that inflammation precedes oligodendrocyte death and neuronal loss and that macrophages/ microglia are the only cells persisting in large numbers in the chronic phase of the disease, probably because of the switch from proinflammatory to anti-inflammatory phenotype.
Keywords
EAE; spinal cord; leukocytes; T cells; macrophages; microglia; oligodendrocytes; neuronal damage; immunohistochemistry
About this articleTitle
Immunohistochemical analysis of spinal cord components in mouse model of experimental autoimmune encephalomyelitis
Journal
Folia Histochemica et Cytobiologica
Issue
Article type
Original paper
Pages
151-158
Published online
2018-08-30
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2947
Article views/downloads
2640
DOI
Pubmed
Bibliographic record
Folia Histochem Cytobiol 2018;56(3):151-158.
Keywords
EAE
spinal cord
leukocytes
T cells
macrophages
microglia
oligodendrocytes
neuronal damage
immunohistochemistry
Authors
Grazyna Pyka-Fosciak
Mariusz Stasiolek
Jan A. Litwin
References (30)- Gold R, Linington C, Lassmann H. Understanding pathogenesis and therapy of multiple sclerosis via animal models: 70 years of merits and culprits in experimental autoimmune encephalomyelitis research. Brain. 2006; 129(Pt 8): 1953–1971.
- Stromnes IM, Goverman JM, Stromnes IM, et al. Active induction of experimental allergic encephalomyelitis. Nat Protoc. 2006; 1(4): 1810–1819.
- Storch MK, Stefferl A, Brehm U, et al. Autoimmunity to myelin oligodendrocyte glycoprotein in rats mimics the spectrum of multiple sclerosis pathology. Brain Pathol. 1998; 8(4): 681–694.
- Kuchroo VK, Anderson AC, Waldner H, et al. T cell response in experimental autoimmune encephalomyelitis (EAE): role of self and cross-reactive antigens in shaping, tuning, and regulating the autopathogenic T cell repertoire. Annu Rev Immunol. 2002; 20: 101–123.
- Trapp BD, Nave KA. Multiple sclerosis: an immune or neurodegenerative disorder? Annu Rev Neurosci. 2008; 31: 247–269.
- Farooqi N, Gran B, Constantinescu CS. Are current disease-modifying therapeutics in multiple sclerosis justified on the basis of studies in experimental autoimmune encephalomyelitis? J Neurochem. 2010; 115(4): 829–844.
- Hooke Laboratories, protocol EK-2110; available at: https://hookelabs.com/protocols/eaeAI_C57BL6.html.
- Codarri L, Greter M, Becher B. Communication between pathogenic T cells and myeloid cells in neuroinflammatory disease. Trends Immunol. 2013; 34(3): 114–119.
- Rangachari M, Kuchroo VK. Using EAE to better understand principles of immune function and autoimmune pathology. J Autoimmun. 2013; 45: 31–39.
- Herrero-Herranz E, Pardo LA, Gold R, et al. Pattern of axonal injury in murine myelin oligodendrocyte glycoprotein induced experimental autoimmune encephalomyelitis: implications for multiple sclerosis. Neurobiol Dis. 2008; 30(2): 162–173.
- Krishnamoorthy G, Lassmann H, Wekerle H, et al. Spontaneous opticospinal encephalomyelitis in a double-transgenic mouse model of autoimmune T cell/B cell cooperation. J Clin Invest. 2006; 116(9): 2385–2392.
- Pöllinger B, Krishnamoorthy G, Berer K, et al. Spontaneous relapsing-remitting EAE in the SJL/J mouse: MOG-reactive transgenic T cells recruit endogenous MOG-specific B cells. J Exp Med. 2009; 206(6): 1303–1316.
- Lisak RP, Nedelkoska L, Benjamins JA, et al. B cells from patients with multiple sclerosis induce cell death via apoptosis in neurons in vitro. J Neuroimmunol. 2017; 309: 88–99.
- Wu F, Cao W, Yang Y, et al. Extensive infiltration of neutrophils in the acute phase of experimental autoimmune encephalomyelitis in C57BL/6 mice. Histochem Cell Biol. 2010; 133(3): 313–322.
- Aubé B, Lévesque SA, Paré A, et al. Neutrophils mediate blood-spinal cord barrier disruption in demyelinating neuroinflammatory diseases. J Immunol. 2014; 193(5): 2438–2454.
- Yi H, Guo C, Yu X, et al. Mouse CD11b+Gr-1+ myeloid cells can promote Th17 cell differentiation and experimental autoimmune encephalomyelitis. J Immunol. 2012; 189(9): 4295–4304.
- Steinbach K, Piedavent M, Bauer S, et al. Neutrophils amplify autoimmune central nervous system infiltrates by maturing local APCs. J Immunol. 2013; 191(9): 4531–4539.
- Ajami B, Bennett J, Krieger C, et al. Infiltrating monocytes trigger EAE progression, but do not contribute to the resident microglia pool. Nature Neuroscience. 2011; 14(9): 1142–1149.
- Liu C, Li Y, Yu J, et al. Targeting the shift from M1 to M2 macrophages in experimental autoimmune encephalomyelitis mice treated with fasudil. PLoS One. 2013; 8(2): e54841.
- Chu F, Shi M, Lang Y, et al. The roles of macrophages and microglia in multiple sclerosis and experimental autoimmune encephalomyelitis. J Neuroimmunol. 2018; 318: 1–7.
- Shin T, Ahn M, Matsumoto Y. Mechanism of experimental autoimmune encephalomyelitis in Lewis rats: recent insights from macrophages. Anat Cell Biol. 2012; 45(3): 141–148.
- Zawadzka M, Rivers LE, Fancy SPJ, et al. CNS-resident glial progenitor/stem cells produce Schwann cells as well as oligodendrocytes during repair of CNS demyelination. Cell Stem Cell. 2010; 6(6): 578–590.
- Kotter MR, Li WW, Zhao C, et al. Myelin impairs CNS remyelination by inhibiting oligodendrocyte precursor cell differentiation. J Neurosci. 2006; 26(1): 328–332.
- Ozawa K, Suchanek G, Breitschopf H, et al. Patterns of oligodendroglia pathology in multiple sclerosis. Brain. 1994; 117(6): 1311–1322.
- Hisahara S, Okano H, Miura M. Caspase-mediated oligodendrocyte cell death in the pathogenesis of autoimmune demyelination. Neurosci Res. 2003; 46(4): 387–397.
- Recks MS, Stormanns ER, Bader J, et al. Early axonal damage and progressive myelin pathology define the kinetics of CNS histopathology in a mouse model of multiple sclerosis. Clin Immunol. 2013; 149(1): 32–45.
- Vogt J, Paul F, Aktas O, et al. Lower motor neuron loss in multiple sclerosis and experimental autoimmune encephalomyelitis. Ann Neurol. 2009; 66(3): 310–322.
- Cunnea P, Mháille ANí, McQuaid S, et al. Expression profiles of endoplasmic reticulum stress-related molecules in demyelinating lesions and multiple sclerosis. Mult Scler. 2011; 17(7): 808–818.
- Kauppinen TM, Swanson RA. Poly(ADP-ribose) polymerase-1 promotes microglial activation, proliferation, and matrix metalloproteinase-9-mediated neuron death. J Immunol. 2005; 174(4): 2288–2296.
- Zhao D, Feng F, Zhao C, et al. Role of perforin secretion from CD8+ T-cells in neuronal cytotoxicity in multiple sclerosis. Neurol Res. 2018; 40(1): 62–67.
