Borreliacidal OspC Antibody Response of Canines with Lyme Disease Differs Significantly from That of Humans with Lyme Disease
Steven D. LovrichRhonda L. La FleurDean A. JobeJennifer JohnsonE. van Meijgaarden KristaRonald F. SchellSteven M. Callister
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Abstract:
Humans reliably produce high concentrations of borreliacidal OspC antibodies specific for the seven C-terminal amino acids shortly after infection with Borrelia burgdorferi. We show that dogs also produce OspC borreliacidal antibodies but that their frequencies, intensities, and antigenicities differ significantly. The findings therefore confirm a major difference between the borreliacidal antibody responses of humans and canines with Lyme disease.Keywords:
Antibody response
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Borrelia Burgdorferi Infection
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Nymphal Ixodes dammini transmitted Borrelia burgdorferi to 1 of 14 rodents exposed for 24 h, 5 of 14 rodents exposed for 48 h, and 13 of 14 rodents exposed for greater than or equal to 72 h. Prompt removal of attached ticks is a prudent public health measure, especially in regions where Lyme disease is endemic.
Tick-borne disease
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The frequency and specificity of antibodies that bind antigens of Borrelia burgdorferi in sera from 200 individuals with no evidence of past or current Lyme disease was determined. Sera were tested for both IgG and IgM antibodies to B. burgdorferi by Western blotting. The non-Lyme serum group included specimens from healthy adults and children in addition to specimens from patients with viral infection and rheumatic diseases. Crossreactive IgG antibodies occurred more frequently than IgM antibodies. The most frequently bound antigens corresponded to 41 kDa and 60 kDa Borrelial components. Of 200 specimens tested, 100 had antibodies that bound at least 1 antigen. Binding to multiple antigens occurred at much lower frequency. Our results indicate that determination of maximum crossreactivity of non-Lyme sera can be used to establish minimum criteria for determining a positive Western blot result for Lyme disease.
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Objective To determine the potential of detection in CSF of specific Borrelia burgdorferi antigen, OspA, as a marker of infection in neurologic Lyme disease and compare this with the detection of antibody. Design CSF from 83 neurologic patients in an area highly endemic for Lyme disease was examined prospectively for (1) OspA by antigen capture ELISA and Western blot employing monoclonal antibodies, and for (2) B burgdorferi antibodies by ELISA. Results Of the 35 of 83 (42%) patients who were positive for OspA antigen in their CSF, 15 (43%) were antigen positive despite being antibody-negative in CSF. Seven of these 15 (47%) had otherwise normal routine CSF analyses. Six of these 15 (40%) patients met strict CDC surveillance criteria for Lyme disease; four (27%) patients had serocon-version coincident with new neurologic problems; and three (20%) with characteristic syndromes for Lyme disease were seronegative, but had complexed antibody to B burgdorferi. The final two patients (13%) were seropositive and had unexplained neurologic problems not characteristic of Lyme disease. ConclusionsB burgdorferi antigen can be detected in CSF that is otherwise normal by conventional methodology, and can be present without positive CSF antibody. Since CSF antigen implies intrathecal seeding of the infection, the diagnosis of neurologic infection by B burgdorferi should not be excluded solely on the basis of normal routine CSF or negative CSF antibody analyses.
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Sera from patients with Lyme disease were evaluated for their ability to kill Borrelia burgdorferi in vivo and in vitro. Separate groups of C3H mice received sera from seropositive humans with early- or late-stage Lyme disease or from seronegative controls. Eighteen to 24 hours after passive transfer of sera, the mice were challenged with 100,000 low-passage B. burgdorferi strain B31 or CA287 organisms. Sera from subjects with late-stage Lyme disease protected the mice against infection after challenge with B. burgdorferi, but sera from subjects with early-stage Lyme disease were not protective. Late-stage sera also inhibited the growth of B. burgdorferi in microcultures on Barbour-Stoenner-Kelly media better than early-stage sera. Immunoblot analysis revealed that the protective properties of late-stage sera were associated with a response of antibodies to multiple proteins. This response included strong reactivity with the outer-surface proteins A and B, which was lacking in early-stage sera.
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Diversity and mutations in the genes for outer surface proteins (Osps) A and B of Borrelia burgdorferi sensu lato (B. burgdorferi), the spirochetal agent of Lyme disease, suggests that a monovalent OspA or OspB vaccine may not provide protection against antigenically variable naturally occurring B. burgdorferi. We now show that OspA or OspB immunizations protect mice from tick-borne infection with heterogeneous B. burgdorferi from different geographic regions. This result is in distinct contrast to in vitro killing analyses and in vivo protection studies using syringe injections of B. burgdorferi as the challenge inoculum. Evaluations of vaccine efficacy against Lyme disease and other vector-borne infections should use the natural mode of transmission and not be predicated on classification systems or assays that do not rely upon the vector to transmit infection.
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We developed an in vitro assay showing that Borrelia burgdorferi organisms were killed by serum from patients with Lyme disease. Twenty of 20 Lyme disease serum samples caused B. burgdorferi killing in a range of 36 to 99% compared with the mean number of viable spirochetes when sera from 10 healthy individuals were used. The percentage of killing of B. burgdorferi increased with convalescent serum from patients with early Lyme disease. The borreliacidal activity was detectable in some sera diluted 640-fold and was abrogated after treatment with anti-human immunoglobulin G. In contrast, pooled or individual normal human serum did not cause a decrease in the number of viable B. burgdorferi. Borreliacidal activity was also not detected in sera from patients with relapsing fever, rocky mountain spotted fever, syphilis, mononucleosis, rheumatoid factor, or DNA antibodies. Our results show that borreliacidal activity can be used as a specific serodiagnostic test for detecting Lyme disease.
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