Human brain microvascular endothelial cell traversal by Borrelia burgdorferi requires calcium signaling.

Neurological manifestations of Lyme disease (or neuroborreliosis) occur variably and while it is clear that Borrelia burgdorferi can invade the nervous system, how it does so is not well understood. Pathogen penetration through the blood brain barrier (BBB) is often influenced by calcium signaling in the endothelial cells triggered by extracellular host-pathogen interactions. We examined the traversal of B. burgdorferi across the human BBB using in vitro model systems constructed of human brain microvascular endothelial cells (HBMEC) grown on Costar Transwell inserts. Pretreatment of the cell monolayers with BAPTA-AM (an intracellular calcium chelator) or phospholipase C (PLC) inhibitor U73122 inhibited B. burgdorferi transmigration. By 5 h, BAPTA-AM significantly inhibited (82-99%; p <0.017) spirochete traversal of HBMEC compared to DMSO controls. Spirochete traversal was almost totally blocked (> or =99%; p <0.017) after pretreatment with the PLC-beta inhibitor U73122 as a result of barrier tightening based on electric cell-substrate impedance sensing (ECIS). The data suggest a role for calcium signaling in CNS spirochete invasion through endothelial cell barriers.

[1]  K. Kim,et al.  Ca2+/calmodulin-dependent invasion of microvascular endothelial cells of human brain by Escherichia coli K1 , 2008, Cell and Tissue Research.

[2]  J. Muldowney,et al.  Acute tissue-type plasminogen activator release in human microvascular endothelial cells: The roles of Gαq, PLC-β, IP3 and 5,6-epoxyeicosatrienoic acid , 2007, Thrombosis and Haemostasis.

[3]  J. Halperin,et al.  The clinical assessment, treatment, and prevention of lyme disease, human granulocytic anaplasmosis, and babesiosis: clinical practice guidelines by the Infectious Diseases Society of America. , 2006, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[4]  J. Scharfstein,et al.  Blood-brain barrier traversal by African trypanosomes requires calcium signaling induced by parasite cysteine protease. , 2006, The Journal of clinical investigation.

[5]  K. Narayan,et al.  Intrathecal antibody production in a mouse model of Lyme neuroborreliosis , 2006, Journal of Neuroimmunology.

[6]  W. Schaper,et al.  H2O2 induces paracellular permeability of porcine brain-derived microvascular endothelial cells by activation of the p44/42 MAP kinase pathway. , 2005, European journal of cell biology.

[7]  A. Agarwal,et al.  PAR1 Is a Matrix Metalloprotease-1 Receptor that Promotes Invasion and Tumorigenesis of Breast Cancer Cells , 2005, Cell.

[8]  J. Dumler,et al.  Borrelia burgdorferi, Host-Derived Proteases, and the Blood-Brain Barrier , 2005, Infection and Immunity.

[9]  S. Ito,et al.  AFRICAN TRYPANOSOME INTERACTIONS WITH AN IN VITRO MODEL OF THE HUMAN BLOOD–BRAIN BARRIER , 2004, The Journal of parasitology.

[10]  J. Radolf,et al.  Experimental Assessment of the Roles of Linear Plasmids lp25 and lp28-1 of Borrelia burgdorferi throughout the Infectious Cycle , 2004, Infection and Immunity.

[11]  A. Steere,et al.  The emergence of Lyme disease. , 2004, The Journal of clinical investigation.

[12]  J. Halperin.  Lyme disease and the peripheral nervous system , 2003, Muscle & nerve.

[13]  M. Balda,et al.  Functional analysis of tight junctions. , 2003, Methods.

[14]  J. Luong,et al.  Insect cell-based impedance biosensors: a novel technique to monitor the toxicity of environmental pollutants , 2003 .

[15]  A. Easton,et al.  Bradykinin increases permeability by calcium and 5-lipoxygenase in the ECV304/C6 cell culture model of the blood–brain barrier , 2002, Brain Research.

[16]  G. Wormser,et al.  Disease severity in a murine model of lyme borreliosis is associated with the genotype of the infecting Borrelia burgdorferi sensu stricto strain. , 2002, The Journal of infectious diseases.

[17]  L. Matrisian,et al.  Matrix metalloproteinases: they're not just for matrix anymore! , 2001, Current opinion in cell biology.

[18]  G. Wormser,et al.  Impact of Genotypic Variation of Borrelia burgdorferi Sensu Stricto on Kinetics of Dissemination and Severity of Disease in C3H/HeJ Mice , 2001, Infection and Immunity.

[19]  H. Lutz,et al.  Molecular Evidence of Coinfection of Ticks with Borrelia burgdorferi Sensu Lato and the Human Granulocytic Ehrlichiosis Agent in Switzerland , 1999, Journal of Clinical Microbiology.

[20]  C. Taylor,et al.  Pharmacological analysis of intracellular Ca2+ signalling: problems and pitfalls. , 1998, Trends in pharmacological sciences.

[21]  F. Gilles,et al.  Selective expression of adhesion molecules on human brain microvascular endothelial cells , 1997, Journal of Neuroimmunology.

[22]  N. Prasadarao,et al.  Bovine brain microvascular endothelial cells transfected with SV40-large T antigen: Development of an immortalized cell line to study pathophysiology of CNS disease , 1997, In Vitro Cellular & Developmental Biology - Animal.

[23]  D. Liveris,et al.  Molecular typing of Borrelia burgdorferi sensu lato by PCR-restriction fragment length polymorphism analysis , 1995, Journal of clinical microbiology.

[24]  A. Barbour.  Isolation and cultivation of Lyme disease spirochetes. , 1984, The Yale journal of biology and medicine.

[25]  A. Steere,et al.  The spirochetal etiology of Lyme disease. , 1983, The New England journal of medicine.

[26]  J. Muldowney,et al.  Acute tissue-type plasminogen activator release in human microvascular endothelial cells: the roles of Galphaq, PLC-beta, IP3 and 5,6-epoxyeicosatrienoic acid. , 2007, Thrombosis and haemostasis.

[27]  K. Kim,et al.  Differential Ca2+ signaling by thrombin and protease-activated receptor-1-activating peptide in human brain microvascular endothelial cells. , 2004, American journal of physiology. Cell physiology.

[28]  K. Kim,et al.  Differential Ca 2 signaling by thrombin and protease-activated receptor-1-activating peptide in human brain microvascular endothelial cells , 2003 .