Function of theBorrelia burgdorferiFtsH Homolog Is Essential for Viability bothIn VitroandIn Vivoand Independent of HflK/C

ABSTRACTIn many bacteria, the FtsH protease and its modulators, HflK and HflC, form a large protein complex that contributes to both membrane protein quality control and regulation of the cellular response to environmental stress. Both activities are crucial to the Lyme disease pathogenBorrelia burgdorferi, which depends on membrane functions, such as motility, protein transport, and cell signaling, to respond to rapid changes in its environment. Using an inducible system, we demonstrate that FtsH production is essential for both mouse and tick infectivity and forin vitrogrowth ofB. burgdorferi. FtsH depletion inB. burgdorfericells resulted in membrane deformation and cell death. Overproduction of the protease did not have any detectable adverse effects onB. burgdorferigrowthin vitro, suggesting that excess FtsH does not proteolytically overwhelm its substrates. In contrast, we did not observe any phenotype for cells lacking the protease modulators HflK and HflC (ΔHflK/C), although we examined morphology, growth rate, growth under stress conditions, and the complete mouse-tick infectious cycle. Our results demonstrate that FtsH provides an essential function in the life cycle of the obligate pathogenB. burgdorferibut that HflK and HflC do not detectably affect FtsH function.IMPORTANCELyme disease is caused byBorrelia burgdorferi, which is maintained in nature in an infectious cycle alternating between small mammals andIxodesticks.B. burgdorferiproduces specific membrane proteins to successfully infect and persist in these diverse organisms. We hypothesized thatB. burgdorferihas a specific mechanism to ensure that membrane proteins are properly folded and biologically active when needed and removed if improperly folded or dysfunctional. Our experiments demonstrate that FtsH, a protease that fulfills this role in other microorganisms, is essential toB. burgdorferiviability. Cells depleted of FtsH do not survive in laboratory culture medium and cannot colonize mice or ticks, revealing an absolute requirement for this protease. However, the loss of two potential modulators of FtsH activity, HflK and HflC, does not detectably affectB. burgdorferiphysiology. Our results provide the groundwork for the identification of FtsH substrates that are critical for the bacterium’s viability.

Download Full-text

Role of the BBA64 Locus of Borrelia burgdorferi in Early Stages of Infectivity in a Murine Model of Lyme Disease

Infection and Immunity ◽

10.1128/iai.01118-07 ◽

2007 ◽

Vol 76 (1) ◽

pp. 391-402 ◽

Cited By ~ 37

Author(s):

Mahulena Maruskova ◽

M. Dolores Esteve-Gassent ◽

Valerie L. Sexton ◽

J. Seshu

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Immunoblot Analysis ◽

Open Reading Frames ◽

Mammalian Host ◽

Environmental Signals ◽

Significant Difference ◽

Linear Plasmid 54

ABSTRACT Borrelia burgdorferi, the causative agent of Lyme disease, undergoes rapid adaptive gene expression in response to environmental signals encountered during different stages of its life cycle in the arthropod vector or the mammalian host. Among all the plasmid-encoded genes of B. burgdorferi, several linear plasmid 54 (lp54)-encoded open reading frames (ORFs) exhibit the greatest differential expression in response to mammalian host-specific temperature, pH, and other uncharacterized signals. These ORFs include members of the paralogous gene family 54 (pgf 54), such as BBA64, BBA65, and BBA66, present on lp54. In an attempt to correlate transcriptional up-regulation of these pgf 54 members to their role in infectivity, we inactivated BBA64 and characterized the phenotype of this mutant both in vitro and in vivo. There were no major differences in the protein profiles between the BBA64 mutant and the control strains, while immunoblot analysis indicated that inactivation of BBA64 resulted in increased levels of BBA65. Moreover, there was no significant difference in the ability of the BBA64 mutant to infect C3H/HeN mice compared to that of its parental or complemented control strains as determined by culturing of viable spirochetes from infected tissues. However, enumeration of spirochetes using quantitative real-time PCR revealed tissue-specific differences, suggesting a minimal role for BBA64 in the survival of B. burgdorferi in select tissues. Infectivity analysis of the BBA64 mutant suggests that B. burgdorferi may utilize multiple determinants to establish infection in mammalian hosts.

Download Full-text

Evaluation of RevA, a Fibronectin-Binding Protein of Borrelia burgdorferi, as a Potential Vaccine Candidate for Lyme Disease

Clinical and Vaccine Immunology ◽

10.1128/cvi.00758-12 ◽

2013 ◽

Vol 20 (6) ◽

pp. 892-899 ◽

Cited By ~ 12

Author(s):

Angela M. Floden ◽

Tammy Gonzalez ◽

Robert A. Gaultney ◽

Catherine A. Brissette

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Passive Immunization ◽

Surface Protein ◽

Surface Expression ◽

Content Type ◽

Good Target ◽

Potential Vaccine ◽

Mammalian Infection

ABSTRACTPrevious studies indicated that the Lyme disease spirocheteBorrelia burgdorferiexpresses the RevA outer surface protein during mammalian infection. As an adhesin that promotes bacterial interaction with fibronectin, RevA appears to be a good target for preventive therapies. RevA proteins are highly conserved across all Lyme borreliae, and antibodies against RevA protein are cross-reactive among RevA proteins from diverse strains. Mice infected withB. burgdorferimounted a rapid IgM response to RevA, followed by a strong IgG response that generally remained elevated for more than 12 months, suggesting continued exposure of RevA protein to the immune system. RevA antibodies were bactericidalin vitro. To evaluate the RevA antigen as a potential vaccine, mice were vaccinated with recombinant RevA and challenged withB. burgdorferiby inoculation with a needle or by a tick bite. Cultured tissues from all treatment groups were positive forB. burgdorferi. Vaccinated animals also appeared to have similar levels ofB. burgdorferiDNA compared to nonvaccinated controls. Despite its antigenicity, surface expression, and the production of bactericidal antibodies against it, RevA does not protect againstBorrelia burgdorferiinfection in a mouse model. However, passive immunization with anti-RevA antibodies did prevent infection, suggesting the possible utility of RevA-based immunotherapeutics or vaccine.

Download Full-text

Efficacy of an Evernimicin (SCH27899) In Vitro and in an Animal Model of Lyme Disease

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.45.3.936-937.2001 ◽

2001 ◽

Vol 45 (3) ◽

pp. 936-937 ◽

Cited By ~ 7

Author(s):

Charles S. Pavia ◽

Gary P. Wormser ◽

John Nowakowski ◽

Anthony Cacciapuoti

Keyword(s):

Animal Model ◽

Lyme Disease ◽

Borrelia Burgdorferi ◽

Antibiotic Treatment ◽

Murine Model ◽

C3h Mice

ABSTRACT The MICs of evernimicin at which 90% of Borrelia burgdorferi patient isolates were inhibited ranged from 0.1 to 0.5 μg/ml. Evernimicin was as effective as ceftriaxone againstB. burgdorferi in a murine model of experimental Lyme disease. As assessed by culturing the urinary bladders of infected C3H mice, no live Borrelia isolates were recoverable following antibiotic treatment.

Download Full-text

Altered Murine Tissue Colonization by Borrelia burgdorferi following Targeted Deletion of Linear Plasmid 17-Carried Genes

Infection and Immunity ◽

10.1128/iai.05984-11 ◽

2012 ◽

Vol 80 (5) ◽

pp. 1773-1782 ◽

Cited By ~ 9

Author(s):

Timothy Casselli ◽

Yvonne Tourand ◽

Troy Bankhead

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Mutant Strain ◽

Genetic Manipulation ◽

Infectious Clone ◽

Linear Plasmid ◽

Peripheral Tissues ◽

Content Type ◽

Immunodeficient Mice

ABSTRACTThe causative agent of Lyme disease,Borrelia burgdorferi, possesses a segmented genome comprised of a single linear chromosome and upwards of 23 linear and circular plasmids. Much of what is known about plasmid-borne genes comes from studying laboratory clones that have spontaneously lost one or more plasmids duringin vitropassage. Some plasmids, including the linear plasmid lp17, are never or rarely reported to be lost during routine culture; therefore, little is known about the requirement of these conserved plasmids for infectivity. In this study, the effects of deleting regions of lp17 were examined bothin vitroandin vivo. A mutant strain lacking the genesbbd16tobbd25showed no deficiency in the ability to establish infection or disseminate to the bloodstream of mice; however, colonization of peripheral tissues was delayed. Despite the ability to colonize ear, heart, and joint tissues, this mutant exhibited a defect in bladder tissue colonization for up to 56 days postinfection. This phenotype was not observed in immunodeficient mice, suggesting that bladder colonization by the mutant strain was inhibited by an adaptive immune-based mechanism. Moreover, the mutant displayed increased expression of outer surface protein Cin vitro, which was correlated with the absence of the genebbd18. To our knowledge, this is the first report involving genetic manipulation of lp17 in an infectious clone ofB. burgdorferiand reveals for the first time the effects of lp17 gene deletion during murine infection by the Lyme disease spirochete.

Download Full-text

The Borrelia burgdorferi Glycosaminoglycan Binding Protein Bgp in the B31 Strain Is Not Essential for Infectivity despite Facilitating Adherence and Tissue Colonization

Infection and Immunity ◽

10.1128/iai.00667-17 ◽

2017 ◽

Vol 86 (2) ◽

Cited By ~ 5

Author(s):

Samantha Schlachter ◽

Janakiram Seshu ◽

Tao Lin ◽

Steven Norris ◽

Nikhat Parveen

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Binding Protein ◽

Mammalian Host ◽

Infection Model ◽

Binding Assays ◽

Content Type ◽

Mouse Infection Model ◽

Successful Infection

ABSTRACTThe Lyme disease-causing organismBorrelia burgdorferiis transmitted into the mammalian host by an infected-tick bite. Successful infection relies on the ability of this extracellular pathogen to persist and colonize different tissues.B. burgdorferiencodes a large number of adhesins that are able to interact with host ligands to facilitate adherence and tissue colonization. Multiple glycosaminoglycan binding proteins present inB. burgdorferioffer a degree of redundancy of function during infection, and this highlights the importance of glycosaminoglycans as host cell receptors for spirochete adherence. Of particular interest in this study isBorreliaglycosaminoglycan binding protein (Bgp), which binds to heparin-related glycosaminoglycans. The properties of abgptransposon mutant and atrans-complemented derivative were compared to those of the wild-typeB. burgdorferiin thein vitrobinding assays and in infection studies using a C3H/HeJ mouse infection model. We determined that the loss of Bgp impairs spirochete adherence, infectivity, and tissue colonization, resulting in a reduction of inflammatory manifestations of Lyme disease. Although Bgp is not essential for infectivity, it is an important virulence factor ofB. burgdorferithat allows adherence and tissue colonization and contributes to disease severity.

Download Full-text

Active and Passive Immunity against Borrelia burgdorferi Decorin Binding Protein A (DbpA) Protects against Infection

Infection and Immunity ◽

10.1128/iai.66.5.2143-2153.1998 ◽

1998 ◽

Vol 66 (5) ◽

pp. 2143-2153 ◽

Cited By ~ 101

Author(s):

Mark S. Hanson ◽

David R. Cassatt ◽

Betty P. Guo ◽

Nita K. Patel ◽

Michael P. McCarthy ◽

...

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Low Dose ◽

Early Stage ◽

Protein A ◽

Sensu Stricto ◽

Passive Immunity ◽

In Vitro Growth

ABSTRACT Borrelia burgdorferi, the spirochete that causes Lyme disease, binds decorin, a collagen-associated extracellular matrix proteoglycan found in the skin (the site of entry for the spirochete) and in many other tissues. Two borrelial adhesins that recognize this proteoglycan, decorin binding proteins A and B (DbpA and DbpB, respectively), have recently been identified. Infection of mice by low-dose B. burgdorferi challenge elicited antibodies against DbpA and DbpB that were sustained at high levels, suggesting that these antigens are expressed in vivo. Scanning immunoelectron microscopy showed that DbpA was surface accessible on intact borreliae. Passive administration of DbpA antiserum protected mice from infection following challenge with heterologous B. burgdorferi sensu stricto isolates, even when serum administration was delayed for up to 4 days after challenge. DbpA is the first antigen target identified that is capable of mediating immune resolution of early, localizedB. burgdorferi infections. DbpA immunization also protected mice from B. burgdorferi challenge; DbpB immunization was much less effective. DbpA antiserum inhibited in vitro growth of manyB. burgdorferi sensu lato isolates of diverse geographic, phylogenetic, and clinical origins. In combination, these findings support a role for DbpA in the immunoprophylaxis of Lyme disease and suggest that DbpA vaccines have the potential to eliminate early-stageB. burgdorferi infections.

Download Full-text

In vitro and in vivo susceptibility of the Lyme disease spirochete, Borrelia burgdorferi, to four antimicrobial agents.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.31.2.164 ◽

1987 ◽

Vol 31 (2) ◽

pp. 164-167 ◽

Cited By ~ 74

Author(s):

R C Johnson ◽

C Kodner ◽

M Russell

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Antimicrobial Agents

Download Full-text

Comparative in vitro and in vivo susceptibilities of the Lyme disease spirochete Borrelia burgdorferi to cefuroxime and other antimicrobial agents.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.34.11.2133 ◽

1990 ◽

Vol 34 (11) ◽

pp. 2133-2136 ◽

Cited By ~ 39

Author(s):

R C Johnson ◽

C B Kodner ◽

P J Jurkovich ◽

J J Collins

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Antimicrobial Agents

Download Full-text

Repurposing Disulfiram (Tetraethylthiuram Disulfide) as a Potential Drug Candidate against Borrelia burgdorferi In Vitro and In Vivo

Antibiotics ◽

10.3390/antibiotics9090633 ◽

2020 ◽

Vol 9 (9) ◽

pp. 633 ◽

Cited By ~ 2

Author(s):

Hari-Hara S. K. Potula ◽

Jahanbanoo Shahryari ◽

Mohammed Inayathullah ◽

Andrey Victorovich Malkovskiy ◽

Kwang-Min Kim ◽

...

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Post Infection ◽

Systemic Disease ◽

Organ Damage ◽

Sensu Stricto ◽

Effector Memory ◽

Drug Candidate

Lyme disease caused by the Borrelia burgdorferi (Bb or B. burgdorferi) is the most common vector-borne, multi-systemic disease in the USA. Although most Lyme disease patients can be cured with a course of the first line of antibiotic treatment, some patients are intolerant to currently available antibiotics, necessitating the development of more effective therapeutics. We previously found several drugs, including disulfiram, that exhibited effective activity against B. burgdorferi. In the current study, we evaluated the potential of repurposing the FDA-approved drug, disulfiram for its borreliacidal activity. Our results indicate disulfiram has excellent borreliacidal activity against both the log and stationary phase B. burgdorferi sensu stricto B31 MI. Treatment of mice with disulfiram eliminated the B. burgdorferi sensu stricto B31 MI completely from the hearts and urinary bladder by day 28 post infection. Moreover, disulfiram-treated mice showed reduced expressions of inflammatory markers, and thus they were protected from histopathology and cardiac organ damage. Furthermore, disulfiram-treated mice showed significantly lower amounts of total antibody titers (IgM and IgG) at day 21 and total IgG2b at day 28 post infection. FACS analysis of lymph nodes revealed a decrease in the percentage of CD19+ B cells and an increase in total percentage of CD3+ T cells, CD3+ CD4+ T helpers, and naive and effector memory cells in disulfiram-treated mice. Together, our findings suggest that disulfiram has the potential to be repurposed as an effective antibiotic for treating Lyme disease.

Download Full-text