Distinct Characteristics of Resistance toBorrelia burgdorferi-Induced Arthritis in C57BL/6N Mice

ABSTRACT Studies of mice infected with Borrelia burgdorferi have indicated that the severity of arthritis is influenced by the genetic composition of the host: the C3H mouse develops severe arthritis while BALB/c and C57BL/6 mice develop mild arthritis. In this study, the effects of increasing infectious dose on the severity of arthritis were determined in these three mouse strains. C3H/He mice developed severe arthritis at all infectious doses, with 100% infection requiring 200 spirochetes. In BALB/cAnN mice, arthritis severity was dependent on infectious dose; symptoms were mild with infection by 200 B. burgdorferi and progressively more severe with increasing infectious dose. Infection of BALB/cAnN mice with 2 × 104 B. burgdorferi resulted in arthritis with severity identical to that in C3H/He mice. Spirochete levels in rear ankle joints of C3H/HeJ and C3H/HeN mice were relatively high, as detected by PCR, and did not increase with infectious dose. Spirochete levels in joints from BALB/cAnN mice increased with increasing infectious dose to levels found in severely arthritic C3H/He mice. Thus, resistance to severe arthritis in BALB/cAnN mice was conditional: it could be overcome by high infectious dose and the arthritis became severe when high levels of B. burgdorferi were present in joints. A unique response to increasing infectious dose was seen in C57BL/6N mice, which displayed mild to moderate arthritis at all doses of B. burgdorferi tested, up to 2 × 105. At all infectious doses, the levels of spirochetes in ankle joints of C57BL/6N mice were high, equivalent to those found in the severely arthritic C3H/He mice. The arthritis observed in infected (C57BL/6N × C3H/HeN)F1 mice was of severity intermediate between those of the two parental strains. The finding that resistance to severe arthritis in C57BL/6N mice could not be overcome by high infectious doses and was independent of spirochete levels in joints suggested that it was mediated by a distinct mechanism from that operating in BALB/cAnN mice.

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Clearance of Borrelia burgdorferi May Not Be Required for Resistance to Experimental Lyme Arthritis

Infection and Immunity ◽

10.1128/iai.66.5.2065-2071.1998 ◽

1998 ◽

Vol 66 (5) ◽

pp. 2065-2071 ◽

Cited By ~ 25

Author(s):

Charles R. Brown ◽

Steven L. Reiner

Keyword(s):

Borrelia Burgdorferi ◽

Inbred Mouse ◽

Experimental Period ◽

Lyme Arthritis ◽

Mouse Strains ◽

Inbred Mouse Strains ◽

Competitive Pcr ◽

Experimental Animals ◽

Severe Arthritis ◽

C3h Mice

ABSTRACT Infection of inbred mouse strains with Borrelia burgdorferi results in the development of experimental Lyme arthritis. The degree of arthritic pathology has been suggested to correlate with the level of spirochete burden within tissues. To investigate this further, we infected resistant DBA/2 (DBA) and susceptible C3H/HeJ (C3H) mice in the hind footpads and monitored arthritis development for 21 days. To quantitate levels of spirochetes within tissues, we created a competitive PCR molecule containing modified ospA and fla gene segments. C3H mice developed severe arthritis of the tibiotarsal joints, while DBA mice developed only mild inflammation throughout the experimental period. At day 21, when the gross size and histologic composition of ankles revealed significant differences in arthritis between the strains, there was little difference in levels of spirochete DNA as determined by competitive PCR. Cultures of ankle tissue at day 21 were also uniformly positive in both C3H and DBA animals and contained relatively similar levels of spirochetes. These results indicate that the presence of spirochetes in the ankles of experimental animals is not sufficient for arthritis development. Since arthritic and nonarthritic animals can harbor relatively equal spirochete burdens yet retain their distinct phenotypic outcomes, an aberrant or overly exuberant immune response may be an additional requirement for pathology in arthritis-prone mice.

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Increasing the Interaction of Borrelia burgdorferi with Decorin Significantly Reduces the 50 Percent Infectious Dose and Severely Impairs Dissemination

Infection and Immunity ◽

10.1128/iai.00560-07 ◽

2007 ◽

Vol 75 (9) ◽

pp. 4272-4281 ◽

Cited By ~ 19

Author(s):

Qilong Xu ◽

Sunita V. Seemanaplli ◽

Kristy McShan ◽

Fang Ting Liang

Keyword(s):

Borrelia Burgdorferi ◽

Severe Combined Immunodeficiency ◽

Protein A ◽

Antigen Expression ◽

In Vitro Assays ◽

Infectious Dose ◽

Surface Antigen Expression ◽

Immunocompetent Mice ◽

Surface Adhesins

ABSTRACT Tight regulation of surface antigenic expression is crucial for the pathogenic strategy of the Lyme disease spirochete, Borrelia burgdorferi. Here, we report the influence of increasing expression of decorin-binding protein A (DbpA), one of the most investigated spirochetal surface adhesins, on the 50% infectious dose (ID50), dissemination, tissue colonization, pathogenicity, and persistence of B. burgdorferi in the murine host. Our in vitro assays showed that increasing DbpA expression dramatically increased the interaction of B. burgdorferi with decorin and sensitivity to growth inhibition/killing by anti-DbpA antibodies; however, this increased interaction did not affect spirochetal growth and replication in the presence of decorin. Increasing DbpA expression significantly reduced ID50 values and severely impaired dissemination in severe combined immunodeficiency (SCID) and immunocompetent mice. During infection of SCID mice, B. burgdorferi with increased DbpA expression was able to effectively colonize heart and skin tissues, but not joint tissues, completely abrogating arthritis virulence. Although increasing DbpA expression did not affect spirochetal persistence in the skin, it diminished the ability of B. burgdorferi to persist in the heart and joint tissues during chronic infection of immunocompetent mice. Taken together, the study highlights the importance of controlling surface antigen expression in the infectivity, dissemination, tissue colonization, pathogenicity, and persistence of B. burgdorferi during mammalian infection.

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Generality of Post-Antimicrobial Treatment Persistence of Borrelia burgdorferi Strains N40 and B31 in Genetically Susceptible and Resistant Mouse Strains

Infection and Immunity ◽

10.1128/iai.00442-19 ◽

2019 ◽

Vol 87 (10) ◽

Cited By ~ 3

Author(s):

Emir Hodzic ◽

Denise M. Imai ◽

Edlin Escobar

Keyword(s):

Mouse Model ◽

Borrelia Burgdorferi ◽

Clinical Relevance ◽

Antimicrobial Agents ◽

Basic Feature ◽

Antimicrobial Treatment ◽

Mouse Strains ◽

Resistant Mouse ◽

Content Type ◽

Disease Resistant

ABSTRACT A basic feature of infection caused by Borrelia burgdorferi, the etiological agent of Lyme borreliosis, is that persistent infection is the rule in its many hosts. The ability to persist and evade host immune clearance poses a challenge to effective antimicrobial treatment. A link between therapy failure and the presence of persister cells has started to emerge. There is growing experimental evidence that viable but noncultivable spirochetes persist following treatment with several different antimicrobial agents. The current study utilized the mouse model to evaluate if persistence occurs following antimicrobial treatment in disease-susceptible (C3H/HeJ [C3H]) and disease-resistant (C57BL/6 [B6]) mouse strains infected with B. burgdorferi strains N40 and B31 and to confirm the generality of this phenomenon, as well as to assess the persisters’ clinical relevance. The status of infection was evaluated at 12 and 18 months after treatment. The results demonstrated that persistent spirochetes remain viable for up to 18 months following treatment, as well as being noncultivable. The phenomenon of persistence in disease-susceptible C3H mice is equally evident in disease-resistant B6 mice and not unique to any particular B. burgdorferi strain. The results also demonstrate that, following antimicrobial treatment, both strains of B. burgdorferi, N40 and B31, lose one or more plasmids. The study demonstrated that noncultivable spirochetes can persist in a host following antimicrobial treatment for a long time but did not demonstrate their clinical relevance in a mouse model of chronic infection. The clinical relevance of persistent spirochetes beyond 18 months following antimicrobial treatment requires further studies in other animal models.

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Infection of Interleukin 17 Receptor A-Deficient C3H Mice with Borrelia burgdorferi Does Not Affect Their Development of Lyme Arthritis and Carditis

Infection and Immunity ◽

10.1128/iai.00533-15 ◽

2015 ◽

Vol 83 (7) ◽

pp. 2882-2888 ◽

Cited By ~ 8

Author(s):

Carrie E. Lasky ◽

Kara E. Jamison ◽

Darcie R. Sidelinger ◽

Carmela L. Pratt ◽

Guoquan Zhang ◽

...

Keyword(s):

Borrelia Burgdorferi ◽

Lyme Borreliosis ◽

Heart Tissue ◽

Lyme Arthritis ◽

Mouse Strains ◽

Interleukin 17 ◽

Content Type ◽

C3h Mice ◽

A Subunit

Recently, a number of studies have reported the presence of interleukin 17 (IL-17) in patients with Lyme disease, and several murine studies have suggested a role for this cytokine in the development of Lyme arthritis. However, the role of IL-17 has not been studied using the experimental Lyme borreliosis model of infection of C3H mice withBorrelia burgdorferi. In the current study, we investigated the role of IL-17 in the development of experimental Lyme borreliosis by infecting C3H mice devoid of the common IL-17 receptor A subunit (IL-17RA) and thus deficient in most IL-17 signaling. Infection of both C3H and C3H IL-17RA−/−mice led to the production of high levels of IL-17 in the serum, low levels in the heart tissue, and no detectable IL-17 in the joint tissue. The development and severity of arthritis and carditis in the C3H IL-17RA−/−mice were similar to what was seen in wild-type C3H mice. In addition, development of antiborrelia antibodies and clearance of spirochetes from tissues were similar for the two mouse strains. These results demonstrate a limited role for IL-17 signaling through IL-17RA in the development of disease following infection of C3H mice withB. burgdorferi.

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Role of Osteopontin in Murine Lyme Arthritis and Host Defense against Borrelia burgdorferi

Infection and Immunity ◽

10.1128/iai.70.3.1372-1381.2002 ◽

2002 ◽

Vol 70 (3) ◽

pp. 1372-1381 ◽

Cited By ~ 17

Author(s):

Melissa R. Potter ◽

Susan R. Rittling ◽

David T. Denhardt ◽

Randall J. Roper ◽

John H. Weis ◽

...

Keyword(s):

Borrelia Burgdorferi ◽

Host Defense ◽

Bacterial Infections ◽

Inbred Mice ◽

Interleukin 10 ◽

Lyme Arthritis ◽

Chromosome 5 ◽

Severe Arthritis ◽

Antibody Isotypes ◽

Deficient Mice

ABSTRACT Several genetic loci in the mouse have been identified that regulate the severity of Lyme arthritis. The region of chromosome 5 including the osteopontin (OPN) gene (Opn) has been identified in intercross populations of C3H/HeN × C57BL/6 and C3H/HeJ × BALB/cAnN mice. OPN is of particular interest as it is involved in the maintenance and remodeling of tissue during inflammation, it regulates production of interleukin-10 (IL-10) and IL-12 (cytokines implicated in Lyme arthritis), it is necessary for host control of certain bacterial infections, and mice displaying different severities of Lyme arthritis possess different alleles of the OPN gene. Macrophages and splenocytes from OPN-deficient mice on mixed C57BL/6J-129S or inbred 129S backgrounds were stimulated with the Pam3Cys modified lipoprotein from Borrelia burgdorferi, OspA. OPN was not required for OspA-induced cytokine production; however, macrophages from 129S-Opn−/− mice displayed a reduced level of IL-10 production. OPN was also not required for resistance to severe arthritis, as B. burgdorferi-infected 129S-Opn−/− mice developed mild arthritis, as did their wild-type littermates. Arthritis was more severe in OPN-deficient mice on the mixed C57BL/6J-129S backgrounds than in inbred mice of either strain. This increase was most likely due to a gene(s) closely linked to Opn on chromosome 5 in conjunction with other randomly assorting genes. Deficiency in OPN did not influence the numbers of spirochetes in tissues from B. burgdorferi-infected mice, indicating OPN is not part of the host defense to this pathogen. Interestingly, there was no alteration in the B. burgdorferi-specific antibody isotypes in OPN-deficient mice, indicating that its effect on helper T-cell responses is not relevant to the host response to B. burgdorferi.

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Conformational Nature of the Borrelia burgdorferi B31 Outer Surface Protein C Protective Epitope

Infection and Immunity ◽

10.1128/iai.67.10.5463-5469.1999 ◽

1999 ◽

Vol 67 (10) ◽

pp. 5463-5469 ◽

Cited By ~ 33

Author(s):

Robert D. Gilmore ◽

M. Lamine Mbow

Keyword(s):

Borrelia Burgdorferi ◽

Outer Surface ◽

Protein C ◽

Surface Protein ◽

Active Immunization ◽

Laboratory Animals ◽

Transmitted Infection ◽

Infectious Dose ◽

Outer Surface Protein ◽

Protective Epitope

ABSTRACT Active immunization with Escherichia coli-expressed recombinant outer surface protein C (OspC) of Borrelia burgdorferi has been demonstrated to confer protection against a tick-transmitted infection on laboratory animals. A previous study in this laboratory showed that OspC antibody raised against a denatured immunogen isolated from B. burgdorferi cells failed to provide protective immunity. Therefore, to determine whether the protective epitope of the recombinant antigen was sensitive to denaturation, recombinant OspC preparations were subjected to heat and chemical treatments prior to animal immunization. Following seroconversion to OspC, the animals were challenged with an infectious dose of B. burgdorferi B31 by tick bite. Whereas mice immunized with a soluble, nondenatured form continued to show protection rates close to 100%, mice that had been immunized with denatured antigen were not protected. Furthermore, mice that were immunized with an insoluble (rather than a soluble), nondenatured form of the recombinant OspC showed a protection rate of only 40%. Protective epitope localization experiments showed that either the amino or the carboxy end of the recombinant protein was required to react with a protective OspC-specific monoclonal antibody. The data from these experiments demonstrate that a conformational organization of the protein is essential for the protective capability of the strain B31 OspC immunogen.

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Modulation of Cytokine Profiles by the Mycoplasma SuperantigenMycoplasma arthritidis Mitogen Parallels Susceptibility to Arthritis Induced by M. arthritidis

Infection and Immunity ◽

10.1128/iai.68.3.1142-1149.2000 ◽

2000 ◽

Vol 68 (3) ◽

pp. 1142-1149 ◽

Cited By ~ 16

Author(s):

Hong-Hua Mu ◽

Allen D. Sawitzke ◽

Barry C. Cole

Keyword(s):

Interleukin 12 ◽

Mouse Strains ◽

Mild Disease ◽

Cytokine Profiles ◽

C Cell ◽

Severe Arthritis

ABSTRACT Mycoplasma arthritidis mitogen (MAM) is a potent superantigen secreted by M. arthritidis, an agent of murine arthritis. Here we compare the abilities of MAM to induce a panel of cytokines in vitro and in vivo in BALB/c and C3H/HeJ mouse strains that differ in susceptibility to mycoplasmal arthritis. Splenocytes from both mouse strains produced high levels of all cytokines by 24 h following in vitro exposure to MAM. No differences in cytokine profiles were seen irrespective of the MAM dose. However, there were striking differences in cytokine profiles present in supernatants of splenocytes that had been collected from mice after intravenous (i.v.) injection of MAM and subsequently rechallenged with MAM in vitro. Splenocytes collected 24 and 72 h after i.v. injection of MAM and challenged in vitro with MAM showed the most marked divergence in the secreted cytokines. Type 1 cytokines were markedly elevated in C3H/HeJ cell supernatants, whereas they were depressed or remained low in BALB/c cell supernatants. In contrast, the levels of type 2 cytokines were all greatly increased in BALB/c cell cultures but were decreased or remained low in C3H/HeJ supernatants. Interleukin-12 mRNA and protein was also markedly elevated in C3H/HeJ mice, as were the levels of immunoglobulin G2a. The data indicate a major skewing in cytokine profiles to a type 1 inflammatory response in C3H/HeJ mice but to a protective type 2 response in BALB/c mice. These cytokine changes appear to be associated with the severe arthritis in C3H/HeJ mice following injection of M. arthritidis in comparison to the mild disease seen in injected BALB/c mice.

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Borrelia burgdorferi basic membrane proteins A and B participate in the genesis of Lyme arthritis

Journal of Experimental Medicine ◽

10.1084/jem.20070962 ◽

2007 ◽

Vol 205 (1) ◽

pp. 133-141 ◽

Cited By ~ 63

Author(s):

Utpal Pal ◽

Penghua Wang ◽

Fukai Bao ◽

Xiuli Yang ◽

Swapna Samanta ◽

...

Keyword(s):

Borrelia Burgdorferi ◽

Host Response ◽

Gene Array ◽

Lyme Arthritis ◽

Differential Analysis ◽

Wild Type ◽

Severe Arthritis ◽

Polymerase Chain ◽

New Strategies

Lyme arthritis results from colonization of joints by Borrelia burgdorferi and the ensuing host response. Using gene array–based differential analysis of B. burgdorferi gene expression and quantitative reverse trancription-polymerase chain reaction, we identified two paralogous spirochete genes, bmpA and bmpB, that are preferentially up-regulated in mouse joints compared with other organs. Transfer of affinity-purified antibodies against either BmpA or BmpB into B. burgdorferi–infected mice selectively reduced spirochete numbers and inflammation in the joints. B. burgdorferi lacking bmpA/B were therefore generated to further explore the role of these proteins in the pathogenesis of Lyme disease. B. burgdorferi lacking bmpA/B were infectious in mice, but unable to persist in the joints, and they failed to induce severe arthritis. Complementation of the mutant spirochetes with a wild-type copy of the bmpA and bmpB genes partially restored the original phenotype. These data delineate a role for differentially produced B. burgdorferi antigens in spirochete colonization of mouse joints, and suggest new strategies for the treatment of Lyme arthritis.

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Neonatal Mice as Models for Cronobacter sakazakii Infection in Infants

Journal of Food Protection ◽

10.4315/0362-028x-72.11.2363 ◽

2009 ◽

Vol 72 (11) ◽

pp. 2363-2367 ◽

Cited By ~ 13

Author(s):

ARENA N. RICHARDSON ◽

SONYA LAMBERT ◽

MARY ALICE SMITH

Keyword(s):

Total Dose ◽

Infant Formula ◽

Mouse Strain ◽

Lethal Dose ◽

Opportunistic Pathogen ◽

Mouse Strains ◽

Powdered Infant Formula ◽

Infant Formulas ◽

Cronobacter Sakazakii ◽

Infectious Dose

Cronobacter sakazakii is an opportunistic pathogen that has been isolated from powdered infant formulas. C. sakazakii infection can result in serious illnesses such as bacteremia, septicemia, meningitis, and death in at-risk infants who are orally fed contaminated reconstituted powdered infant formulas. The objective of this study was to compare the susceptibilities of BALB/c, C57BL/6, and CD-1 mice to C. sakazakii strain MNW2. We acquired timed-pregnant CD-1 mice and allowed them to give birth naturally. On postnatal day 3.5, each pup was administered a total dose of approximately 102 to 1011 CFU C. sakazakii strain MNW2 in reconstituted powdered infant formula. Mice were observed twice daily for morbidity and mortality. At postnatal day 10.5, the remaining pups were euthanized, and brain, liver, and cecum were excised and analyzed for the presence of C. sakazakii. C. sakazakii was isolated from brains, livers, and ceca in all three mouse strains. The CD-1 mouse strain was the most susceptible of the three, with the lowest infectious dose (102 CFU) and the lowest lethal dose (also 102 CFU).

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