The MEK/ERK pathway is the primary conduit for Borrelia burgdorferi-induced inflammation and P53-mediated apoptosis in oligodendrocytes
Lyme neuroborreliosis (LNB) affects both the central and peripheral nervous systems. In a rhesus macaque model of LNB we had previously shown that brains of rhesus macaques inoculated with Borrelia burgdorferi release inflammatory mediators, and undergo oligodendrocyte and neuronal cell death. In vitro analysis of this phenomenon indicated that while B. burgdorferi can induce inflammation and apoptosis of oligodendrocytes per se, microglia are required for neuronal apoptosis. We hypothesized that the inflammatory milieu elicited by the bacterium in microglia or oligodendrocytes contributes to the apoptosis of neurons and glial cells, respectively, and that downstream signaling events in NFkB and/or MAPK pathways play a role in these phenotypes. To test these hypotheses in oligodendrocytes, several pathway inhibitors were used to determine their effect on inflammation and apoptosis, as induced by B. burgdorferi. In a human oligodendrocyte cell line (MO3.13), inhibition of the ERK pathway in the presence of B. burgdorferi markedly reduced inflammation, followed by the JNK, p38 and NFkB pathway inhibition. In addition to eliciting inflammation, B. burgdorferi also increased total p53 protein levels, and suppression of the ERK pathway mitigated this effect. While inhibition of p53 had a minimal effect in reducing inflammation, suppression of the ERK pathway or p53 reduced apoptosis as measured by active caspase-3 activity and the TUNEL assay. A similar result was seen in primary human oligodendrocytes wherein suppression of ERK or p53 reduced apoptosis. It is possible that inflammation and apoptosis in oligodendrocytes are divergent arms of MAPK pathways, particularly the MEK/ERK pathway.
KeywordsBorrelia burgdorferi Oligodendrocytes Inflammation Apoptosis MAPK
This study was supported by the National Institute of Neurologic Disorders and Stroke through Grant NS048952, and by the National Center for Research Resources/Office of Research Infrastructure Programs of the National Institutes of Health through Grant P51RR000164/P51OD011104. We thank the TNPRC Pathogen Detection and Quantification Core Laboratory for help with the multiplex ELISA assays. Ms. Robin Rodriguez of the TNPRC Media Laboratory is gratefully acknowledged for her assistance with graphics.
Conflict of interest
The authors declare that they have no conflict of interest.
- 6.Benach JL, Garcia-Monco JC (1992) Aspects of the pathogenesis of neuroborreliosis. In: Schutzer S (ed) Lyme disease: molecular and immunological approaches. Cold Spring Harbor Laboratory Press, Plainview, New York, pp 1–10Google Scholar
- 7.Philipp MT, Aydintug MK, Bohm RP Jr, Cogswell FB, Dennis VA, Lanners HN, Lowrie RC Jr, Roberts ED, Conway MD, Karacorlu M, Peyman GA, Gubler DJ, Johnson BJ, Piesman J, Gu Y (1993) Early and early disseminated phases of Lyme disease in the rhesus monkey: a model for infection in humans. Infect Immun 61(7):3047–3059PubMedCentralPubMedGoogle Scholar
- 12.Straubinger RK, Straubinger AF, Harter L, Jacobson RH, Chang YF, Summers BA, Erb HN, Appel MJ (1997) Borrelia burgdorferi migrates into joint capsules and causes an up-regulation of interleukin-8 in synovial membranes of dogs experimentally infected with ticks. Infect Immun 65(4):1273–1285PubMedCentralPubMedGoogle Scholar
- 13.Burns MJ, Sellati TJ, Teng EI, Furie MB (1997) Production of interleukin-8 (IL-8) by cultured endothelial cells in response to Borrelia burgdorferi occurs independently of secreted [corrected] IL-1 and tumor necrosis factor alpha and is required for subsequent transendothelial migration of neutrophils. Infect Immun 65(4):1217–1222PubMedCentralPubMedGoogle Scholar
- 16.Kondrusik M, Swierzbinska R, Pancewicz S, Zajkowska J, Grygorczuk S, Hermanowska-Szpakowicz T (2004) Evaluation of proinflammatory cytokine (TNF-alpha, IL-1beta, IL-6, IFN-gamma) concentrations in serum and cerebrospinal fluid of patients with neuroborreliosis. Neurol Neurochir Pol 38(4):265–270PubMedGoogle Scholar
- 17.Grygorczuk S, Pancewicz S, Zajkowska J, Kondrusik M, Rwierzbinska R, Hermanowska-Szpakowicz T (2004) Concentrations of macrophage inflammatory proteins MIP-1alpha and MIP-1beta and interleukin 8 (il-8) in lyme borreliosis. Infection 32(6):350–355. doi: 10.1007/s15010-004-3110-4 PubMedCrossRefGoogle Scholar
- 18.Grygorczuk S, Zajkowska J, Swierzbinska R, Pancewicz S, Kondrusik M, Hermanowska-Szpakowicz T (2005) Concentration of interferon-inducible T cell chemoattractant and monocyte chemotactic protein-1 in serum and cerebrospinal fluid of patients with Lyme borreliosis. Rocz Akad Med Bialymst 50:173–178PubMedGoogle Scholar
- 24.Ramesh G, Borda JT, Dufour J, Kaushal D, Ramamoorthy R, Lackner AA, Philipp MT (2008) Interaction of the Lyme disease spirochete Borrelia burgdorferi with brain parenchyma elicits inflammatory mediators from glial cells as well as glial and neuronal apoptosis. Am J Pathol 173(5):1415–1427. doi: 10.2353/ajpath.2008.080483 PubMedCrossRefGoogle Scholar
- 36.Sadik CD, Hunfeld KP, Bachmann M, Kraiczy P, Eberhardt W, Brade V, Pfeilschifter J, Muhl H (2008) Systematic analysis highlights the key role of TLR2/NF-kappaB/MAP kinase signaling for IL-8 induction by macrophage-like THP-1 cells under influence of Borrelia burgdorferi lysates. Int J Biochem Cell Biol 40(11):2508–2521. doi: 10.1016/j.biocel.2008.04.014 PubMedCrossRefGoogle Scholar
- 38.Izadi H, Motameni AT, Bates TC, Olivera ER, Villar-Suarez V, Joshi I, Garg R, Osborne BA, Davis RJ, Rincon M, Anguita J (2007) c-Jun N-terminal kinase 1 is required for Toll-like receptor 1 gene expression in macrophages. Infect Immun 75(10):5027–5034. doi: 10.1128/IAI.00492-07 PubMedCentralPubMedCrossRefGoogle Scholar
- 39.Olson CM, Hedrick MN, Izadi H, Bates TC, Olivera ER, Anguita J (2007) p38 mitogen-activated protein kinase controls NF-kappaB transcriptional activation and tumor necrosis factor alpha production through RelA phosphorylation mediated by mitogen- and stress-activated protein kinase 1 in response to Borrelia burgdorferi antigens. Infect Immun 75(1):270–277. doi: 10.1128/IAI.01412-06 PubMedCentralPubMedCrossRefGoogle Scholar
- 40.Hawley K, Navasa N, Olson CM Jr, Bates TC, Garg R, Hedrick MN, Conze D, Rincon M, Anguita J (2012) Macrophage p38 mitogen-activated protein kinase activity regulates invariant natural killer T-cell responses during Borrelia burgdorferi infection. J Infect Dis 206(2):283–291. doi: 10.1093/infdis/jis332 PubMedCrossRefGoogle Scholar
- 49.Strom E, Sathe S, Komarov PG, Chernova OB, Pavlovska I, Shyshynova I, Bosykh DA, Burdelya LG, Macklis RM, Skaliter R, Komarova EA, Gudkov AV (2006) Small-molecule inhibitor of p53 binding to mitochondria protects mice from gamma radiation. Nat Chem Biol 2(9):474–479. doi: 10.1038/nchembio809 PubMedCrossRefGoogle Scholar
- 52.Pelaia G, Cuda G, Vatrella A, Grembiale RD, De Sarro G, Maselli R, Costanzo FS, Avvedimento VE, Rotiroti D, Marsico SA (2001) Effects of glucocorticoids on activation of c-jun N-terminal, extracellular signal-regulated, and p38 MAP kinases in human pulmonary endothelial cells. Biochem Pharmacol 62(12):1719–1724PubMedCrossRefGoogle Scholar