Neuropathogenicity of non-viable Borrelia burgdorferi ex vivo

AbstractEven after appropriate treatment, a proportion of Lyme disease patients suffer from a constellation of symptoms, collectively called Post-Treatment Lyme Disease Syndrome (PTLDS). Brain PET scan of patients with PTLDS have demonstrated likely glial activation indicating persistent neuroinflammatory processes. It is possible that unresolved bacterial remnants can continue to cause neuroinflammation. In previous studies, we have shown that non-viable Borrelia burgdorferi can induce neuroinflammation and apoptosis in an oligodendrocyte cell line. In this follow-up study, we analyze the effect of sonicated remnants of B. burgdorferi on primary rhesus frontal cortex (FC) and dorsal root ganglion (DRG) explants. Five FC and three DRG tissue fragments from rhesus macaques were exposed to sonicated B. burgdorferi and analyzed for 26 inflammatory mediators. Live bacteria and medium alone served as positive and negative control, respectively. Tissues were also analyzed for cell types mediating inflammation and overall apoptotic changes. Non-viable B. burgdorferi induced significant levels of several inflammatory mediators in both FC and DRG, similar to live bacteria. However, the levels induced by non-viable B. burgdorferi was often (several fold) higher than those induced by live ones, especially for IL-6, CXCL8 and CCL2. This effect was also more profound in the FC than in the DRG. Although the levels often differed, both live and dead fragments induced the same mediators, with significant overlap between FC and DRG. In the FC, immunohistochemical staining for several inflammatory mediators showed the presence of multiple mediators in astrocytes, followed by microglia and oligodendrocytes, in response to bacterial remnants. Staining was also seen in endothelial cells. In the DRG, chemokine/cytokine staining was predominantly seen in S100 positive (glial) cells. B. burgdorferi remnants also induced significant levels of apoptosis in both the FC and DRG. Apoptosis was confined to S100 + cells in the DRG while distinct neuronal apoptosis was also detected in most FC tissues in response to sonicated bacteria. Non-viable B. burgdorferi can continue to be neuropathogenic to both CNS and PNS tissues with effects likely more profound in the former. Persistence of remnant-induced neuroinflammatory processes can lead to long term health consequences.

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Fibroblast Protection of Borrelia burgdorferi from Doxycycline, Cefuroxime and Daptomycin Combination is Eliminated by Oregano or Carvacrol Essential Oil

10.1101/861575 ◽

2019 ◽

Author(s):

Chunxiang Bai ◽

Hua Yang ◽

Peng Cui ◽

Rong Quan ◽

Ying Zhang

Keyword(s):

Lyme Disease ◽

Essential Oil ◽

Borrelia Burgdorferi ◽

Drug Combination ◽

Defense Mechanisms ◽

Ex Vivo ◽

Fibroblast Cells ◽

Murine Fibroblast ◽

Drug Regimens

AbstractBorrelia burgdorferi could be occasionally recovered from patients after antibiotic treatment, which indicates it may resist eradication by antibiotic and host defense mechanisms. Skin fibroblast cells have previously been shown to protect the killing of B. burgdorferi by ceftriaxone, a powerful antibiotic commonly used to treat Lyme disease. In this study, we evaluated if fibroblast cells could also protect against the doxycycline+ cefuroxime+ daptomycin drug combination which has previously been shown to completely eradicate highly persistent biofilm-like microcolonies of B. burgdorferi. To do so, we utilized a GFP-labeled B. burgdorferi for infection of murine fibroblast cells and assessed the effect of the drug combination on killing the bacteria in the presence or absence of the fibroblast cells. Surprisingly, we found that fibroblasts could protect B. burgdorferi from being completely killed by the drug combination doxycycline, cefuroxime and daptomycin, which eradicated B. burgdorferi completely in the absence of fibroblast cells. Interestingly, addition of essential oil carvacrol or oregano at 0.1% could enhance the activity of the doxycycline+ cefuroxime+ daptomycin drug combination and led to complete eradication of B. burgdorferi even in the presence of fibroblast cells. Further studies are needed to determine if the essential oil drug combinations could eradicate persistent B. burgdorferi infection in vivo in animal models. Our study provides a useful and convenient ex vivo model for evaluating different drug regimens needed for developing more effective treatment of persistent Lyme disease in the future.

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Early Disseminated Lyme Disease with Carditis Complicated by Posttreatment Lyme Disease Syndrome

Case Reports in Infectious Diseases ◽

10.1155/2017/5847156 ◽

2017 ◽

Vol 2017 ◽

pp. 1-4

Author(s):

Cheryl Novak ◽

Andrew Harrison ◽

John Aucott

Keyword(s):

Infectious Disease ◽

Nervous System ◽

Lyme Disease ◽

Borrelia Burgdorferi ◽

Cardiac Involvement ◽

Conduction System

Lyme disease is an infectious disease caused by the bacterium Borrelia burgdorferi. When untreated, infection may spread to the heart, nervous system, and joints. Cardiac involvement usually manifests as abnormalities of the conduction system and bradycardia. Treatment of Lyme disease is generally effective, with a subset of patients experiencing persistent, sometimes long-term symptoms called posttreatment Lyme disease syndrome.

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Type I IFN is siloed in endosomes

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1921324117 ◽

2020 ◽

Vol 117 (30) ◽

pp. 17510-17512 ◽

Cited By ~ 1

Author(s):

Jennie B. Altman ◽

Justin Taft ◽

Tim Wedeking ◽

Conor N. Gruber ◽

Michael Holtmannspötter ◽

...

Keyword(s):

Ex Vivo ◽

Cell Types ◽

Specific Cell ◽

Type I ◽

Type I Ifn ◽

Long Term Effects ◽

Negative Regulators ◽

Downstream Signaling

Type I IFN (IFN-I) is thought to be rapidly internalized and degraded following binding to its receptor and initiation of signaling. However, many studies report the persistent effects mediated by IFN-I for days or even weeks, both ex vivo and in vivo. These long-lasting effects are attributed to downstream signaling molecules or induced effectors having a long half-life, particularly in specific cell types. Here, we describe a mechanism explaining the long-term effects of IFN-I. Following receptor binding, IFN-I is siloed into endosomal compartments. These intracellular “IFN silos” persist for days and can be visualized by fluorescence and electron microscopy. However, they are largely dormant functionally, due to IFN-I−induced negative regulators. By contrast, in individuals lacking these negative regulators, such as ISG15 or USP18, this siloed IFN-I can continue to signal from within the endosome. This mechanism may underlie the long-term effects of IFN-I therapy and may contribute to the pathophysiology of type I interferonopathies.

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Borrelia burgdorferi infection induces long-term memory-like responses in macrophages with tissue-wide consequences in the heart

PLoS Biology ◽

10.1371/journal.pbio.3001062 ◽

2021 ◽

Vol 19 (1) ◽

pp. e3001062

Author(s):

Diego Barriales ◽

Itziar Martín-Ruiz ◽

Ana Carreras-González ◽

Marta Montesinos-Robledo ◽

Mikel Azkargorta ◽

...

Keyword(s):

Borrelia Burgdorferi ◽

Inflammatory Responses ◽

Long Term Memory ◽

Term Memory ◽

Lyme Carditis ◽

Extracutaneous Manifestation ◽

Live Bacteria

Lyme carditis is an extracutaneous manifestation of Lyme disease characterized by episodes of atrioventricular block of varying degrees and additional, less reported cardiomyopathies. The molecular changes associated with the response to Borrelia burgdorferi over the course of infection are poorly understood. Here, we identify broad transcriptomic and proteomic changes in the heart during infection that reveal a profound down-regulation of mitochondrial components. We also describe the long-term functional modulation of macrophages exposed to live bacteria, characterized by an augmented glycolytic output, increased spirochetal binding and internalization, and reduced inflammatory responses. In vitro, glycolysis inhibition reduces the production of tumor necrosis factor (TNF) by memory macrophages, whereas in vivo, it produces the reversion of the memory phenotype, the recovery of tissue mitochondrial components, and decreased inflammation and spirochetal burdens. These results show that B. burgdorferi induces long-term, memory-like responses in macrophages with tissue-wide consequences that are amenable to be manipulated in vivo.

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Coexposure to Borrelia burgdorferi and Babesia microti Does Not Worsen the Long-Term Outcome of Lyme Disease

Clinical Infectious Diseases ◽

10.1086/317465 ◽

2000 ◽

Vol 31 (5) ◽

pp. 1149-1154 ◽

Cited By ~ 33

Author(s):

T. J. Wang ◽

M. H. Liang ◽

O. Sangha ◽

C. B. Phillips ◽

R. A. Lew ◽

...

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Babesia Microti ◽

Term Outcome ◽

Long Term Outcome

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Direct demonstration of antigenic substitution of Borrelia burgdorferi ex vivo: exploration of the paradox of the early immune response to outer surface proteins A and C in Lyme disease.

Journal of Experimental Medicine ◽

10.1084/jem.183.1.261 ◽

1996 ◽

Vol 183 (1) ◽

pp. 261-269 ◽

Cited By ~ 141

Author(s):

R R Montgomery ◽

S E Malawista ◽

K J Feen ◽

L K Bockenstedt

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Outer Surface ◽

Ex Vivo ◽

Surface Proteins ◽

Rt Pcr ◽

Variable Expression ◽

Outer Surface Proteins ◽

Direct Demonstration

The outer surface proteins (Osps) of Borrelia burgdorferi, the etiologic agent of Lyme disease, are principle targets of protective immune responses against this organism. Whereas most North American strains of B. burgdorferi in culture express an abundant amount of Osp A, antibodies to this protein are either absent or only weakly detected in the sera of naturally infected patients or experimentally infected mice. In contrast, Osp C, which has variable expression on cultured organisms; elicits an early, strong humoral response. To examine this paradox, we have studied the in vivo adaptation of a cloned population of B. burgdorferi strain N40 during the early course of experimental murine borreliosis. As in human disease, antibodies to Osp A were only weakly present in the early immune repertoire after murine inoculation with low dose (10(3)) spirochetes. In contrast, antibodies to Osp C were prominent, even though on cultured spirochetes Osp C mRNA and protein expression could not be detected by reverse transcription polymerase chain reaction (RT-PCR) or indirect immunofluorescence, respectively. These observations led us to investigate the expression of Osp A and Osp C in vivo. By direct fluorescent staining of uncultured spirochetes ex vivo and by PCR amplification of spirochetal mRNA, we show that Osp C is indeed expressed by some spirochetes after infection in the mouse. Spirochetes expressing Osp A could also be detected within the first 2 wk of infection, but not at 30 d. Osp A mRNA, although present at day 14 of infection, could not be amplified by RT-PCR at day 30, suggesting that the expression of this Osp is transient. This further implies that the late burst in Osp A antibodies in both mice and humans may be anamnestic. These results indicate that either Osp C is upregulated on spirochetes after infection, or Osp C-expressing spirochetes expand preferentially over those expressing Osp A during infection. These results have important implications for vaccine design and offer one explanation for the failure of Osp A antibodies to eradicate spirochetes from the infected host.

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ELISA-Based Measurement of Antibody Responses and PCR-Based Detection Profiles Can Distinguish between Active Infection and Early Clearance ofBorrelia burgdorferi

Clinical and Developmental Immunology ◽

10.1155/2012/138069 ◽

2012 ◽

Vol 2012 ◽

pp. 1-8 ◽

Cited By ~ 11

Author(s):

John J. Lazarus ◽

Akisha L. McCarter ◽

Kari Neifer-Sadhwani ◽

R. Mark Wooten

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Intradermal Injection ◽

Antibody Responses ◽

Productive Infection ◽

Active Infection ◽

Bacterial Dna ◽

Large Numbers ◽

Live Bacteria ◽

Established Infection

Borrelia burgdorferiis a spirochetal bacterium that causes Lyme disease. These studies address whether current research methods using either ELISA to detect seroconversion toB. burgdorferiantigens or PCR quantification of bacterial DNA within tissues can accurately distinguish between a productive infection versus aB. burgdorferiexposure that is rapidly cleared by the innate responses. Mice receiving even minimal doses of liveB. burgdorferiproduced significantly moreB. burgdorferi-specific IgM and IgG than groups receiving large inocula of heat-killed bacteria. Additionally, sera from mice injected with varied doses of killedB. burgdorferirecognized unique borrelial antigens compared to mice infected with liveB. burgdorferi. Intradermal injection of killedB. burgdorferiresulted in rapid DNA clearance from skin, whereas DNA was consistently detected in skin inoculated with viableB. burgdorferi. These data indicate that both ELISA-based serological analyses and PCR-based methods of assessingB. burgdorferiinfection clearly distinguish between an established infection with live bacteria and exposure to large numbers of bacteria that are promptly cleared by the innate responses.

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Bovine Leukemia Virus-Induced Lymphocytosis and Increased Cell Survival Mainly Involve the CD11b+B-Lymphocyte Subset in Sheep

Journal of Virology ◽

10.1128/jvi.72.5.4413-4420.1998 ◽

1998 ◽

Vol 72 (5) ◽

pp. 4413-4420 ◽

Cited By ~ 21

Author(s):

Nathalie Chevallier ◽

Madeleine Berthelemy ◽

Danielle Le Rhun ◽

Véronique Lainé ◽

Daniel Levy ◽

...

Keyword(s):

Bovine Leukemia Virus ◽

B Lymphocyte ◽

Ex Vivo ◽

Leukemia Virus ◽

Cell Types ◽

Persistent Lymphocytosis ◽

Viral Expression ◽

Protein Kinase C Inhibitor

ABSTRACT In this study, we show that bovine leukemia virus (BLV)-induced persistent lymphocytosis (PL) results from the in vivo expansion of the CD11b+ B-lymphocyte population. This subset shares phenotypic characteristics with murine and human B-1 cells. BLV interactions with the sheep B-1-like subset were explored. We found that B-1- and B-2-like cells are initially infected to similar extents. However, in long-term-infected sheep, the viral load is higher in B-1-like cells and only B-1- and not B-2-like cells show increased ex vivo survival compared to that in uninfected sheep. Ex vivo viral expression was found in both B-1- and B-2-like cells, indicating that both cell types support viral replication. Finally, cycloheximide and a protein kinase C inhibitor (H7) that blocks the ex vivo activation of viral expression did not affect the increased survival in B-1-like cells, suggesting that resistance to apoptosis is acquired in vivo. Collectively, these results indicate a peculiar susceptibility of sheep B-1-like cells to BLV transforming effects and further support the involvement of increased survival in BLV pathogenesis.

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Detection of Immune Complexes Is Not Independent of Detection of Antibodies in Lyme Disease Patients and Does Not Confirm Active Infection with Borrelia burgdorferi

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.12.9.1036-1040.2005 ◽

2005 ◽

Vol 12 (9) ◽

pp. 1036-1040 ◽

Cited By ~ 14

Author(s):

Adriana R. Marques ◽

Ronald L. Hornung ◽

Len Dally ◽

Mario T. Philipp

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Rhesus Macaques ◽

Protein A ◽

Surface Protein ◽

Laboratory Diagnosis ◽

Enzyme Linked Immunosorbent Assay ◽

Active Infection ◽

Serum Samples ◽

Outer Surface Protein A

ABSTRACT The Borrelia burgdorferi-specific immune complex (IC) test, which uses polyethylene glycol (PEG) precipitation to isolate ICs from serum, has been used as a research test in the laboratory diagnosis of early Lyme disease (LD) and has been proposed as a marker of active infection. We examined whether B. burgdorferi-specific antibodies were present within PEG-precipitated ICs (PEG-ICs) in patients with LD, posttreatment Lyme disease syndrome, and controls, including individuals who received the outer surface protein A (OspA) vaccine. Using a B. burgdorferi whole-cell enzyme-linked immunosorbent assay (ELISA), we obtained positive PEG-IC results not only in patients with a history of LD, but also in individuals vaccinated with OspA vaccine. The frequency of positive PEG-IC ELISAs in OspA vaccinees was significantly higher with ELISA-reactive than with ELISA-negative unprocessed serum samples (P = 0.001), demonstrating dependency between the tests. Similar results were found using samples from rhesus macaques infected with B. burgdorferi, uninfected macaques vaccinated with OspA, and controls. Therefore, testing for the presence of antibodies against B. burgdorferi in PEG-IC preparations is not more likely to reflect active infection than testing in unprocessed serum and should not be used in individuals who received the OspA vaccine.

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Live Attenuated Borrelia burgdorferi Targeted Mutants in an Infectious Strain Background Protect Mice from Challenge Infection

Clinical and Vaccine Immunology ◽

10.1128/cvi.00302-16 ◽

2016 ◽

Vol 23 (8) ◽

pp. 725-731 ◽

Cited By ~ 5

Author(s):

Beth L. Hahn ◽

Lavinia J. Padmore ◽

Laura C. Ristow ◽

Michael W. Curtis ◽

Jenifer Coburn

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Parental Strain ◽

Content Type ◽

Protective Antigens ◽

Homologous Strain ◽

Live Bacteria ◽

At Risk Populations ◽

Mammalian Infection

ABSTRACTBorrelia burgdorferi,B. garinii, andB. afzeliiare all agents of Lyme disease in different geographic locations. If left untreated, Lyme disease can cause significant and long-term morbidity, which may continue after appropriate antibiotic therapy has been administered and live bacteria are no longer detectable. The increasing incidence and geographic spread of Lyme disease are renewing interest in the vaccination of at-risk populations. We took the approach of vaccinating mice with two targeted mutant strains ofB. burgdorferithat, unlike the parental strain, are avirulent in mice. Mice vaccinated with both strains were protected against a challenge with the parental strain and a heterologousB. burgdorferistrain by either needle inoculation or tick bite. In ticks, the homologous strain was eliminated but the heterologous strain was not, suggesting that the vaccines generated a response to antigens that are produced by the bacteria both early in mammalian infection and in the tick. Partial protection againstB. gariniiinfection was also conferred. Protection was antibody mediated, and reactivity to a variety of proteins was observed. These experiments suggest that live attenuatedB. burgdorferistrains may be informative regarding the identification of protective antigens produced by the bacteria and recognized by the mouse immune systemin vivo. Further work may illuminate new candidates that are effective and safe for the development of Lyme disease vaccines.

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