Recombinant or Peptide Antigens in the Serology of Lyme Arthritis in Children

Heikkilä, Tero T.; Huppertz, Hans-Iko; Seppälä, Ilkka; Sillanpää, Heidi; Saxén, Harri; Lahdenne, Pekka

doi:10.1086/375371

Cited by 35 publications

(35 citation statements)

References 27 publications

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“…Recombinant DbpA preparations obtained from different B. burgdorferi sensu lato species have been evaluated in Europe to detect antibodies in patients with LB (61,122,124). These recombinants detect mostly IgG antibodies and have the greatest sensitivities (93 to 100%) in sera from patients with neuroborreliosis and late stages of LB.…”

Section: Immunologic Diagnosis Of B Burgdorferi Sensu Lato Infectionmentioning

confidence: 99%

Diagnosis of Lyme Borreliosis

et al. 2005

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A large amount of knowledge has been acquired since the original descriptions of Lyme borreliosis (LB) and of its causative agent, Borrelia burgdorferi sensu stricto. The complexity of the organism and the variations in the clinical manifestations of LB caused by the different B. burgdorferi sensu lato species were not then anticipated. Considerable improvement has been achieved in detection of B. burgdorferi sensu lato by culture, particularly of blood specimens during early stages of disease. Culturing plasma and increasing the volume of material cultured have accomplished this. Further improvements might be obtained if molecular methods are used for detection of growth in culture and if culture methods are automated. Unfortunately, culture is insensitive in extracutaneous manifestations of LB. PCR and culture have high sensitivity on skin samples of patients with EM whose diagnosis is based mostly on clinical recognition of the lesion. PCR on material obtained from extracutaneous sites is in general of low sensitivity, with the exception of synovial fluid. PCR on synovial fluid has shown a sensitivity of up to >90% (when using four different primer sets) in patients with untreated or partially treated Lyme arthritis, making it a helpful confirmatory test in these patients. Currently, the best use of PCR is for confirmation of the clinical diagnosis of suspected Lyme arthritis in patients who are IgG immunoblot positive. PCR should not be used as the sole laboratory modality to support a clinical diagnosis of extracutaneous LB. PCR positivity in seronegative patients suspected of having late manifestations of LB most likely represents a false-positive result. Because of difficulties in direct methods of detection, laboratory tests currently in use are mainly those detecting antibodies to B. burgdorferi sensu lato. Tests used to detect antibodies to B. burgdorferi sensu lato have evolved from the initial formats as more knowledge on the immunodominant antigens has been collected. The recommendation for two-tier testing was an attempt to standardize testing and improve specificity in the United States. First-tier assays using whole-cell sonicates of B. burgdorferi sensu lato need to be standardized in terms of antigen composition and detection threshold of specific immunoglobulin classes. The search for improved serologic tests has stimulated the development of recombinant protein antigens and the synthesis of specific peptides from immunodominant antigens. The use of these materials alone or in combination as the source of antigen in a single-tier immunoassay may someday replace the currently recommended two-tier testing strategy. Evaluation of these assays is currently being done, and there is evidence that certain of these antigens may be broadly cross-reactive with the B. burgdorferi sensu lato species causing LB in Europe

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Section: Immunologic Diagnosis Of B Burgdorferi Sensu Lato Infectionmentioning

confidence: 99%

Diagnosis of Lyme Borreliosis

et al. 2005

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“…vlsE variation has been shown to occur within 4 days of experimental infection of mice with B. burgdorferi B31 and continues throughout the course of infection but has not been observed in vitro or in the tick vector (21, 53, 54). The conservation of vls sequences in other strains and species of Borrelia indicate that the vls locus is important for the life cycle of Lyme disease agents (23,25,49).Lyme disease patients mount a robust antibody response directed towards VlsE (29,33), and patient sera have been shown to react strongly with the IR6 invariable region of the protein (1,16,28,31,32,39,40,44). With experimentally infected mice, Triton X-114 extraction studies indicate that VlsE is present at high levels in joint and ear tissues but not in heart tissue (9), suggesting differential expression.…”

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confidence: 99%

“…Lyme disease patients mount a robust antibody response directed towards VlsE (29,33), and patient sera have been shown to react strongly with the IR6 invariable region of the protein (1,16,28,31,32,39,40,44). With experimentally infected mice, Triton X-114 extraction studies indicate that VlsE is present at high levels in joint and ear tissues but not in heart tissue (9), suggesting differential expression.…”

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confidence: 99%

Effects of vlsE Complementation on the Infectivity of Borrelia burgdorferi Lacking the Linear Plasmid lp28-1

2004

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The loss of linear plasmid lp28-1, which contains the vls antigenic variation locus, is associated with reduced infectivity of Borrelia burgdorferi in immunocompetent mice. The recombinant shuttle vector pBBE22, which includes the virulence determinant BBE22 from lp25 and restores infectivity to readily transformable B. burgdorferi lacking lp25 and lp56, was used to determine the effect of trans expression of vlsE on virulence. Spirochetes lacking lp28-1 were complemented with the plasmid pBBE22:vlsE, containing both BBE22 and vlsE. VlsE protein produced by this construct was expressed and surface accessible in in vitro-cultured B. burgdorferi, as determined by surface proteolysis and immunoblot analysis. Clones lacking lp25 but containing lp28-1 and either pBBE22 or pBBE22:vlsE were reisolated consistently from immunocompetent mice 8 weeks after infection. In contrast, a clone lacking both lp25 and lp28-1 and complemented with pBBE22:vlsE was isolated from only a single tissue of one of six C3H/HeN mice 8 weeks postinfection. These results indicate that either an intact vls antigenic variation locus or another determinant on lp28-1 is required to restore complete infectivity. In addition, an isogenic clone that retained lp28-1 was complemented with the vlsE shuttle plasmid and was examined for vlsE sequence variation and infectivity. Sequence variation was not observed for the shuttle plasmid, indicating that the cis arrangement of vlsE and the vls silent cassettes in lp28-1 facilitate vlsE gene conversion. Lack of vlsE sequence variation on the shuttle plasmid thus did not result in clearance of the trans-complemented strain in immunocompetent mice under the conditions tested.Lyme borreliosis, the most prevalent vector-borne disease in the United States, is a chronic infection caused by Borrelia burgdorferi and other members of the genus Borrelia (6). Spirochetes are transmitted to mammalian hosts by Ixodes ticks, leading to the development of an annular rash called erythema migrans at the site of inoculation and progressing to a multisystemic infection with neurological, arthritic, and cardiac manifestations (45). As infection advances and Borrelia disseminate into deeper tissues in the host, a strong immune response is elicited towards the pathogen, including the development of Borrelia-specific antibodies (7,8,11,14,17). Though an active immune response develops early during infection, Borrelia is able to escape clearance and persist for months to years. Elucidation of the mechanisms of immune evasion may lead to a better understanding of the pathobiology of Lyme disease.The vls (Vmp-like sequence) locus of B. burgdorferi B31 is on the linear plasmid lp28-1, a plasmid associated with infectivity in the mouse model (26, 27, 42, 52). The vls locus consists of an expression site (vlsE) and 15 unexpressed (silent) vls cassettes. The silent cassettes have high homology to the central cassette region of vlsE. Within the cassettes, there are six variable regions interspersed between highly conserved regions (52). ...

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“…To identify Bb epitopes, in addition to OspA [165][166][167][168][169][170][171][172][173] , with an antibiotic-refractory associated DRB binding profile, 17 known immunogenic Bb proteins (Akin et al, 2001;Bacon et al, 2003;Dressler et al, 1993;Heikkila et al, 2003;Salazar et al, 2005) (Table 1) were screened with the bioinformatics program TEPITOPE. This program scans proteins for peptides predicted to bind one or more of 25 different HLA-DR molecules, and it ranks peptide binding according to stringency (threshold parameter), which can be set anywhere from 1 (the most stringent) to 10 (the least stringent).…”

Section: Bb T Cell Epitopes Predicted To Have An Antibiotic-refractormentioning

confidence: 99%

Searching for borrelial T cell epitopes associated with antibiotic-refractory Lyme arthritis

Drouin

Glickstein

Kwok

et al. 2008

Molecular Immunology

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Antibiotic-refractory Lyme arthritis is believed to result from an infection-induced autoimmune response triggered by the spirochete Borrelia burgdorferi (Bb). Disease susceptibility is associated with the HLA alleles DRB1*0101, 0401, 0402, 0404, 0405 and DRB5*0101, and all these MHC molecules bind the Bb epitope OspA 163-175 . However, not all patients have a proliferative response to this epitope. To identify other possible Bb epitopes involved in this disease process, the algorithm TEPITOPE was used to scan 17 immunogenetic Bb proteins for potential T cell epitopes with a refractory arthritis-associated MHC binding profile, and the Bb proteome was searched for peptides with sequence homology to OspA [165][166][167][168][169][170][171][172][173] . Sixteen promising T epitopes were identified and their MHC binding profiles to 13 MHC molecules were verified using in vitro MHC/peptide binding assays. One peptide, BBK32 392-404 , had a strong refractory-arthritis associated MHC binding profile, and another GK 297-306 shared sequence homology to OspA [165][166][167][168][169][170][171][172][173] . However, patient cells did not proliferate in response to either peptide making it highly unlikely they were involved in a refractory course. A comparison of the in silico and in vitro results revealed that TEPITOPE correctly predicted 74% of the in vitro binding peptides, but it incorrectly predicted that 44% of the in vitrononbinding peptides would bind. For a particular MHC molecule, concordance between the in silico and in vitro results varied anywhere between 33% and 100%. Therefore, while additional Bb epitopes may be involved in the development of antibiotic-refractory Lyme arthritis, recognition of OspA [163][164][165][166][167][168][169][170][171][172][173][174][175] remains the only known Bb epitope associated with this disease.

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Recombinant or Peptide Antigens in the Serology of Lyme Arthritis in Children

Cited by 35 publications

References 27 publications

Diagnosis of Lyme Borreliosis

Diagnosis of Lyme Borreliosis

Effects of vlsE Complementation on the Infectivity of Borrelia burgdorferi Lacking the Linear Plasmid lp28-1

Searching for borrelial T cell epitopes associated with antibiotic-refractory Lyme arthritis

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