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Open Access 01-12-2016 | Research

Complement system activation contributes to the ependymal damage induced by microbial neuraminidase

Authors: Pablo Granados-Durán, María Dolores López-Ávalos, Timothy R. Hughes, Krista Johnson, B. Paul Morgan, Paul P. Tamburini, Pedro Fernández-Llebrez, Jesús M. Grondona

Published in: Journal of Neuroinflammation | Issue 1/2016

Abstract

Background

In the rat brain, a single intracerebroventricular injection of neuraminidase from Clostridium perfringens induces ependymal detachment and death. This injury occurs before the infiltration of inflammatory blood cells; some reports implicate the complement system as a cause of these injuries. Here, we set out to test the role of complement.

Methods

The assembly of the complement membrane attack complex on the ependymal epithelium of rats injected with neuraminidase was analyzed by immunohistochemistry. Complement activation, triggered by neuraminidase, and the participation of different activation pathways were analyzed by Western blot. In vitro studies used primary cultures of ependymal cells and explants of the septal ventricular wall. In these models, ependymal cells were exposed to neuraminidase in the presence or absence of complement, and their viability was assessed by observing beating of cilia or by trypan blue staining. The role of complement in ependymal damage induced by neuraminidase was analyzed in vivo in two rat models of complement blockade: systemic inhibition of C5 by using a function blocking antibody and testing in C6-deficient rats.

Results

The complement membrane attack complex immunolocalized on the ependymal surface in rats injected intracerebroventricularly with neuraminidase. C3 activation fragments were found in serum and cerebrospinal fluid of rats treated with neuraminidase, suggesting that neuraminidase itself activates complement. In ventricular wall explants and isolated ependymal cells, treatment with neuraminidase alone induced ependymal cell death; however, the addition of complement caused increased cell death and disorganization of the ependymal epithelium. In rats treated with anti-C5 and in C6-deficient rats, intracerebroventricular injection of neuraminidase provoked reduced ependymal alterations compared to non-treated or control rats. Immunohistochemistry confirmed the absence of membrane attack complex on the ependymal surfaces of neuraminidase-exposed rats treated with anti-C5 or deficient in C6.

Conclusions

These results demonstrate that the complement system contributes to ependymal damage and death caused by neuraminidase. However, neuraminidase alone can induce moderate ependymal damage without the aid of complement.

Bruni JE. Ependymal development, proliferation, and functions: a review. Microsc Res Tech. 1998;41(1):2–13.CrossRefPubMed

Kuchler S, Graff MN, Gobaille S, Vincendon G, Roche AC, Delaunoy JP, et al. Mannose dependent tightening of the rat ependymal cell barrier. In vivo and in vitro study using neoglycoproteins. Neurochem Int. 1994;24(1):43–55.CrossRefPubMed

Rodriguez EM, Blazquez JL, Pastor FE, Pelaez B, Pena P, Peruzzo B, et al. Hypothalamic tanycytes: a key component of brain-endocrine interaction. Int Rev Cytol. 2005;247:89–164. doi:10.1016/S0074-7696(05)47003-5.CrossRefPubMed

Doetsch F, Garcia-Verdugo JM, Alvarez-Buylla A. Cellular composition and three-dimensional organization of the subventricular germinal zone in the adult mammalian brain. J Neurosci. 1997;17(13):5046–61.PubMed

Lim DA, Tramontin AD, Trevejo JM, Herrera DG, Garcia-Verdugo JM, Alvarez-Buylla A. Noggin antagonizes BMP signaling to create a niche for adult neurogenesis. Neuron. 2000;28(3):713–26.CrossRefPubMed

Coskun V, Wu H, Blanchi B, Tsao S, Kim K, Zhao J, et al. CD133+ neural stem cells in the ependyma of mammalian postnatal forebrain. Proc Natl Acad Sci U S A. 2008;105(3):1026–31.CrossRefPubMedPubMedCentral

Paez P, Batiz LF, Roales-Bujan R, Rodriguez-Perez LM, Rodriguez S, Jimenez AJ, et al. Patterned neuropathologic events occurring in hyh congenital hydrocephalic mutant mice. J Neuropathol Exp Neurol. 2007;66(12):1082–92. doi:10.1097/nen.0b013e31815c1952.CrossRefPubMed

Jimenez AJ, Garcia-Verdugo JM, Gonzalez CA, Batiz LF, Rodriguez-Perez LM, Paez P, et al. Disruption of the neurogenic niche in the subventricular zone of postnatal hydrocephalic hyh mice. J Neuropathol Exp Neurol. 2009;68(9):1006–20. doi:10.1097/NEN.0b013e3181b44a5a.CrossRefPubMed

Granados-Duran P, Lopez-Avalos MD, Grondona JM, Gomez-Roldan Mdel C, Cifuentes M, Perez-Martin M, et al. Neuroinflammation induced by intracerebroventricular injection of microbial neuraminidase. Front Med. 2015;2:14. doi:10.3389/fmed.2015.00014.CrossRef

10.

Johnson RT, Johnson KP, Edmonds CJ. Virus-induced hydrocephalus: development of aqueductal stenosis in hamsters after mumps infection. Science. 1967;157(3792):1066–7.CrossRefPubMed

11.

Johnson RT, Johnson KP. Hydrocephalus as a sequela of experimental myxovirus infections. Exp Mol Pathol. 1969;10(1):68–80.CrossRefPubMed

12.

Margolis G, Kilham L. Hydrocephalus in hamsters, ferrets, rats, and mice following inoculations with reovirus type I. II. Pathologic studies. Lab Invest. 1969;21(3):189–98.PubMed

13.

Takano T, Mekata Y, Yamano T, Shimada M. Early ependymal changes in experimental hydrocephalus after mumps virus inoculation in hamsters. Acta Neuropathol. 1993;85(5):521–5.CrossRefPubMed

14.

Marrie RA, Wolfson C. Multiple sclerosis and Epstein-Barr virus. The Canadian Journal of Infectious Diseases = J canadien des Mal Infect. 2002;13(2):111–8.CrossRef

15.

Ascherio A. Epstein-Barr virus in the development of multiple sclerosis. Expert Rev Neurother. 2008;8(3):331–3. doi:10.1586/14737175.8.3.331.CrossRefPubMed

16.

Ebert T, Kotler M. Prenatal exposure to influenza and the risk of subsequent development of schizophrenia. Isr Med Assoc J. 2005;7(1):35–8.PubMed

17.

Fatemi SH, Reutiman TJ, Folsom TD, Huang H, Oishi K, Mori S, et al. Maternal infection leads to abnormal gene regulation and brain atrophy in mouse offspring: implications for genesis of neurodevelopmental disorders. Schizophr Res. 2008;99(1-3):56–70. doi:10.1016/j.schres.2007.11.018.CrossRefPubMedPubMedCentral

18.

Schneider-Schaulies J, Meulen V, Schneider-Schaulies S. Measles infection of the central nervous system. J Neurovirol. 2003;9(2):247–52. doi:10.1080/13550280390193993.CrossRefPubMed

19.

Rima BK, Duprex WP. Molecular mechanisms of measles virus persistence. Virus Res. 2005;111(2):132–47. doi:10.1016/j.virusres.2005.04.005.CrossRefPubMed

20.

Lucas SM, Rothwell NJ, Gibson RM. The role of inflammation in CNS injury and disease. Br J Pharmacol. 2006;147 Suppl 1:S232–40. doi:10.1038/sj.bjp.0706400.PubMedPubMedCentral

21.

Grondona JM, Perez-Martin M, Cifuentes M, Perez J, Jimenez AJ, Perez-Figares JM, et al. Ependymal denudation, aqueductal obliteration and hydrocephalus after a single injection of neuraminidase into the lateral ventricle of adult rats. J Neuropathol Exp Neurol. 1996;55(9):999–1008.CrossRefPubMed

22.

Kornfeld R, Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–64. doi:10.1146/annurev.bi.54.070185.003215.CrossRefPubMed

23.

Muller-Eberhard HJ. The membrane attack complex of complement. Annu Rev Immunol. 1986;4:503–28. doi:10.1146/annurev.iy.04.040186.002443.CrossRefPubMed

24.

Stahel PF, Morganti-Kossmann MC, Kossmann T. The role of the complement system in traumatic brain injury. Brain Res Brain Res Rev. 1998;27(3):243–56.CrossRefPubMed

25.

Gasque P, Dean YD, McGreal EP, VanBeek J, Morgan BP. Complement components of the innate immune system in health and disease in the CNS. Immunopharmacology. 2000;49(1-2):171–86.CrossRefPubMed

26.

Casarsa C, De Luigi A, Pausa M, De Simoni MG, Tedesco F. Intracerebroventricular injection of the terminal complement complex causes inflammatory reaction in the rat brain. Eur J Immunol. 2003;33(5):1260–70. doi:10.1002/eji.200323574.CrossRefPubMed

27.

Canova C, Neal JW, Gasque P. Expression of innate immune complement regulators on brain epithelial cells during human bacterial meningitis. J Neuroinflammation. 2006;3:22. doi:10.1186/1742-2094-3-22.CrossRefPubMedPubMedCentral

28.

Lauf PK. Immunological and physiological characteristics of the rapid immune hemolysis of neuraminidase-treated sheep red cells produced by fresh guinea pig serum. J Exp Med. 1975;142(4):974–88.CrossRefPubMed

29.

Pangburn MK. Host recognition and target differentiation by factor H, a regulator of the alternative pathway of complement. Immunopharmacology. 2000;49(1-2):149–57.CrossRefPubMed

30.

Pangburn MK, Pangburn KL, Koistinen V, Meri S, Sharma AK. Molecular mechanisms of target recognition in an innate immune system: interactions among factor H, C3b, and target in the alternative pathway of human complement. J Immunol. 2000;164(9):4742–51.CrossRefPubMed

31.

Donin N, Jurianz K, Ziporen L, Schultz S, Kirschfink M, Fishelson Z. Complement resistance of human carcinoma cells depends on membrane regulatory proteins, protein kinases and sialic acid. Clin Exp Immunol. 2003;131(2):254–63.CrossRefPubMedPubMedCentral

32.

Marques MB, Kasper DL, Pangburn MK, Wessels MR. Prevention of C3 deposition by capsular polysaccharide is a virulence mechanism of type III group B streptococci. Infect Immun. 1992;60(10):3986–93.PubMedPubMedCentral

33.

Fujita T, Ohi H, Endo M, Ohsawa I, Kanmatsuse K. The role of sialidase in the development of hypocomplementemia in postinfectious acute glomerulonephritis. Clin Immunol. 1999;92(1):97–102. doi:10.1006/clim.1999.4729.CrossRefPubMed

34.

Ramaglia V, King RH, Nourallah M, Wolterman R, de Jonge R, Ramkema M, et al. The membrane attack complex of the complement system is essential for rapid wallerian degeneration. J Neurosci. 2007;27(29):7663–72. doi:10.1523/JNEUROSCI.5623-06.2007.CrossRefPubMed

35.

Mead RJ, Singhrao SK, Neal JW, Lassmann H, Morgan BP. The membrane attack complex of complement causes severe demyelination associated with acute axonal injury. J Immunol. 2002;168(1):458–65.CrossRefPubMed

36.

Graves IL. Heat inactivation of the neuraminidase and haemagglutinin estimated in the agglutination-separation reactions using red blood cells sensitized with Newcastle disease virus. Vet Res. 1999;30(4):383–92.PubMed

37.

Rother RP, Arp J, Jiang J, Ge W, Faas SJ, Liu W, et al. C5 blockade with conventional immunosuppression induces long-term graft survival in presensitized recipients. Am J Transplant Off J Am Soc Transplant Am Soc Transplant Surg. 2008;8(6):1129–42. doi:10.1111/j.1600-6143.2008.02222.x.CrossRef

38.

Rollins SA, Matis LA, Springhorn JP, Setter E, Wolff DW. Monoclonal antibodies directed against human C5 and C8 block complement-mediated damage of xenogeneic cells and organs. Transplantation. 1995;60(11):1284–92.CrossRefPubMed

39.

Costa C, Zhao L, Shen Y, Su X, Hao L, Colgan SP, et al. Role of complement component C5 in cerebral ischemia/reperfusion injury. Brain Res. 2006;1100(1):142–51. doi:10.1016/j.brainres.2006.05.029.CrossRefPubMed

40.

Grondona JM, Granados-Duran P, Fernandez-Llebrez P, Lopez-Avalos MD. A simple method to obtain pure cultures of multiciliated ependymal cells from adult rodents. Histochem Cell Biol. 2013;139(1):205–20. doi:10.1007/s00418-012-1008-2.CrossRefPubMed

41.

Heijnen BH, Straatsburg IH, Padilla ND, Van Mierlo GJ, Hack CE, Van Gulik TM. Inhibition of classical complement activation attenuates liver ischaemia and reperfusion injury in a rat model. Clin Exp Immunol. 2006;143(1):15–23. doi:10.1111/j.1365-2249.2005.02958.x.CrossRefPubMedPubMedCentral

42.

Ricklin D, Hajishengallis G, Yang K, Lambris JD. Complement: a key system for immune surveillance and homeostasis. Nat Immunol. 2010;11(9):785–97. doi:10.1038/ni.1923.CrossRefPubMedPubMedCentral

43.

Podack ER, Tschoop J, Müller-Eberhard HJ. Molecular organization of C9 within the membrane attack complex of complement. Induction of circular C9 polymerization by the C5b-8 assembly. J Exp Med. 1982;156(1):268–82.CrossRefPubMedPubMedCentral

44.

Laursen SB, Thiel S, Teisner B, Holmskov U, Wang Y, Sim RB, et al. Bovine conglutinin binds to an oligosaccharide determinant presented by iC3b, but not by C3, C3b or C3c. Immunology. 1994;81(4):648–54.PubMedPubMedCentral

45.

Orth D, Ehrlenbach S, Brockmeyer J, Khan AB, Huber G, Karch H, et al. EspP, a serine protease of enterohemorrhagic escherichia coli, impairs complement activation by cleaving complement factors C3/C3b and C5. Infect Immun. 2010;78(10):4294–301. doi:10.1128/IAI.00488-10.CrossRefPubMedPubMedCentral

46.

Spiller OB, Morgan BP. Antibody-independent activation of the classical complement pathway by cytomegalovirus-infected fibroblasts. J Infect Dis. 1998;178(6):1597–603.CrossRefPubMed

47.

Grondona JM, Perez-Martin M, Cifuentes M, Perez J, Estivill-Torrus G, Perez-Figares JM, et al. Neuraminidase injected into the cerebrospinal fluid impairs the assembly of the glycoproteins secreted by the subcommissural organ preventing the formation of Reissner’s fiber. Histochem Cell Biol. 1998;109(4):391–8.CrossRefPubMed

48.

Pérez-Martín M. Papel del epitelio ependimario en la estabilidad de los ventrículos cerebrales y en la neurogénesis en el adulto. Doctoral Thesis. Universidad de Málaga; 2000.

49.

Gomez-Roldan MC, Perez-Martin M, Capilla-Gonzalez V, Cifuentes M, Perez J, Garcia-Verdugo JM, et al. Neuroblast proliferation on the surface of the adult rat striatal wall after focal ependymal loss by intracerebroventricular injection of neuraminidase. J Comp Neurol. 2008;507(4):1571–87. doi:10.1002/cne.21618.CrossRef

50.

Carbonell WS, Murase SI, Horwitz AF, Mandell JW. Infiltrative microgliosis: activation and long-distance migration of subependymal microglia following periventricular insults. J Neuroinflammation. 2005;2(1):5. doi:10.1186/1742-2094-2-5.CrossRefPubMedPubMedCentral

51.

Tompkins WA, Seth P, Gee S, Rawls WE. Neuraminidase reversal of resistance to lysis of herpes simplex virus-infected cells by antibody and complement. J Immunol. 1976;116(2):489–95.PubMed

52.

Barnum SR. Complement biosynthesis in the central nervous system. Crit Rev Oral Biol Med. 1995;6(2):132–46.CrossRefPubMed

53.

Stahel PF, Barnum SR. Bacterial meningitis: complement gene expression in the central nervous system. Immunopharmacology. 1997;38(1-2):65–72.CrossRefPubMed

54.

Morgan BP, Gasque P, Singhrao S, Piddlesden SJ. The role of complement in disorders of the nervous system. Immunopharmacology. 1997;38(1-2):43–50.CrossRefPubMed

55.

Meri S, Morgan BP, Davies A, Daniels RH, Olavesen MG, Waldmann H, et al. Human protectin (CD59), an 18,000-20,000 MW complement lysis restricting factor, inhibits C5b-8 catalysed insertion of C9 into lipid bilayers. Immunology. 1990;71(1):1–9.PubMedPubMedCentral

56.

Rollins SA, Sims PJ. The complement-inhibitory activity of CD59 resides in its capacity to block incorporation of C9 into membrane C5b-9. J Immunol. 1990;144(9):3478–83.PubMed

57.

Kazatchkine MD, Fearon DT, Austen KF. Human alternative complement pathway: membrane-associated sialic acid regulates the competition between B and beta1 H for cell-bound C3b. J Immunol. 1979;122(1):75–81.PubMed

58.

Pangburn MK, Müller-Eberhard HJ. Complement C3 convertase: cell surface restriction of beta1H control and generation of restriction on neuraminidase-treated cells. Proc Natl Acad Sci U S A. 1978;75(5):2416–20.CrossRefPubMedPubMedCentral

59.

Ninomiya H, Stewart BH, Rollins SA, Zhao J, Bothwell AL, Sims PJ. Contribution of the N-linked carbohydrate of erythrocyte antigen CD59 to its complement-inhibitory activity. J Biol Chem. 1992;267(12):8404–10.PubMed

60.

Medicus RG, Götze O, Müller-Eberhard HJ. Alternative pathway of complement: recruitment of precursor properdin by the labile C3/C5 convertase and the potentiation of the pathway. J Exp Med. 1976;144(4):1076–93.CrossRefPubMed

61.

Rawal N, Pangburn MK. Role of the C3b-binding site on C4b-binding protein in regulating classical pathway C5 convertase. Mol Immunol. 2007;44(6):1105–14. doi:10.1016/j.molimm.2006.07.282.CrossRefPubMed

62.

Petit L, Gibert M, Popoff MR. Clostridium perfringens: toxinotype and genotype. Trends Microbiol. 1999;7(3):104–10.CrossRefPubMed

63.

Li J, Sayeed S, Robertson S, Chen J, McClane BA. Sialidases affect the host cell adherence and epsilon toxin-induced cytotoxicity of Clostridium perfringens type D strain CN3718. PLoS Pathog. 2011;7(12):e1002429. doi:10.1371/journal.ppat.1002429.CrossRefPubMedPubMedCentral

64.

Vakeva A, Meri S. Complement activation and regulator expression after anoxic injury of human endothelial cells. APMIS. 1998;106(12):1149–56.CrossRefPubMed

65.

Peckham RM, Handrigan MT, Bentley TB, Falabella MJ, Chrovian AD, Stahl GL, et al. C5-blocking antibody reduces fluid requirements and improves responsiveness to fluid infusion in hemorrhagic shock managed with hypotensive resuscitation. J Appl Physiol. 2007;102(2):673–80. doi:10.1152/japplphysiol.00917.2006.CrossRefPubMed

66.

Bhole D, Stahl GL. Molecular basis for complement component 6 (C6) deficiency in rats and mice. Immunobiology. 2004;209(7):559–68. doi:10.1016/j.imbio.2004.08.001.CrossRefPubMed

67.

Leenaerts PL, Stad RK, Hall BM, Van Damme BJ, Vanrenterghem Y, Daha MR. Hereditary C6 deficiency in a strain of PVG/c rats. Clin Exp Immunol. 1994;97(3):478–82.CrossRefPubMedPubMedCentral

68.

van Dixhoorn MG, Timmerman JJ, Van Gijlswijk-Janssen DJ, Muizert Y, Verweij C, Discipio RG, et al. Characterization of complement C6 deficiency in a PVG/c rat strain. Clin Exp Immunol. 1997;109(2):387–96.CrossRefPubMed

69.

Molgaard CA, Gresham L. Swine flu vaccine and amyotrophic lateral sclerosis. JAMA. 1986;255(17):2294.CrossRefPubMed

70.

Takahashi M, Yamada T. A possible role of influenza a virus infection for Parkinson’s disease. Adv Neurol. 2001;86:91–104.PubMed

71.

Finsterer J, Hess B. Neuromuscular and central nervous system manifestations of Clostridium perfringens infections. Infection. 2007;35(6):396–405. doi:10.1007/s15010-007-6345-z.CrossRefPubMed

72.

Alexander JJ, Anderson AJ, Barnum SR, Stevens B, Tenner AJ. The complement cascade: yin-yang in neuroinflammation—neuro-protection and -degeneration. J Neurochem. 2008;107(5):1169–87. doi:10.1111/j.1471-4159.2008.05668.x.CrossRefPubMedPubMedCentral

73.

Brennan FH, Anderson AJ, Taylor SM, Woodruff TM, Ruitenberg MJ. Complement activation in the injured central nervous system: another dual-edged sword? J Neuroinflammation. 2012;9:137. doi:10.1186/1742-2094-9-137.CrossRefPubMedPubMedCentral

74.

Melis JP, Strumane K, Ruuls SR, Beurskens FJ, Schuurman J, Parren PW. Complement in therapy and disease: regulating the complement system with antibody-based therapeutics. Mol Immunol. 2015;67(2 Pt A):117–30. doi:10.1016/j.molimm.2015.01.028.CrossRefPubMed

75.

Morgan BP, Harris CL. Complement, a target for therapy in inflammatory and degenerative diseases. Nat Rev Drug Discov. 2015;14(12):857–77. doi:10.1038/nrd4657.CrossRefPubMed

76.

Stephan AH, Barres BA, Stevens B. The complement system: an unexpected role in synaptic pruning during development and disease. Annu Rev Neurosci. 2012;35:369–89. doi:10.1146/annurev-neuro-061010-113810.CrossRefPubMed

77.

Schafer DP, Lehrman EK, Kautzman AG, Koyama R, Mardinly AR, Yamasaki R, et al. Microglia sculpt postnatal neural circuits in an activity and complement-dependent manner. Neuron. 2012;74(4):691–705. doi:10.1016/j.neuron.2012.03.026.CrossRefPubMedPubMedCentral

78.

Hong S, Beja-Glasser VF, Nfonoyim BM, Frouin A, Li S, Ramakrishnan S et al. Complement and microglia mediate early synapse loss in Alzheimer mouse models. Science. 2016. doi:10.1126/science.aad8373.

79.

Shi Q, Colodner KJ, Matousek SB, Merry K, Hong S, Kenison JE, et al. Complement C3-deficient mice fail to display age-related hippocampal decline. J Neurosci. 2015;35(38):13029–42. doi:10.1523/JNEUROSCI.1698-15.2015.CrossRefPubMed

Title: Complement system activation contributes to the ependymal damage induced by microbial neuraminidase
Authors: Pablo Granados-Durán
María Dolores López-Ávalos
Timothy R. Hughes
Krista Johnson
B. Paul Morgan
Paul P. Tamburini
Pedro Fernández-Llebrez
Jesús M. Grondona
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: Journal of Neuroinflammation / Issue 1/2016
Electronic ISSN: 1742-2094
DOI: https://doi.org/10.1186/s12974-016-0576-9

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Complement system activation contributes to the ependymal damage induced by microbial neuraminidase

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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