Borrelia burgdorferi erp proteins are immunogenic in mammals infected by tick bite, and their synthesis is inducible in cultured bacteria - PubMed (original) (raw)

Borrelia burgdorferi erp proteins are immunogenic in mammals infected by tick bite, and their synthesis is inducible in cultured bacteria

B Stevenson et al. Infect Immun. 1998 Jun.

Abstract

Borrelia burgdorferi, the causative agent of Lyme disease, can contain multiple genes encoding different members of the Erp lipoprotein family. Some arthropod-borne bacteria increase the synthesis of proteins required for transmission or mammalian infection when cultures are shifted from cool, ambient air temperature to a warmer, blood temperature. We found that all of the erp genes known to be encoded by infectious isolate B31 were differentially expressed in culture after a change in temperature, with greater amounts of message being produced by bacteria shifted from 23 to 35 degrees C than in those maintained at 23 degrees C. Mice infected with B31 by tick bite produced antibodies that recognized each of the Erp proteins within 4 weeks of infection, suggesting that the Erp proteins are produced by the bacteria during the early stages of mammalian infection and may play roles in transmission from ticks to mammals. Several of the B31 Erp proteins were also recognized by antibodies from patients with Lyme disease and may prove to be useful antigens for diagnostic testing or as components of a protective vaccine.

PubMed Disclaimer

Figures

FIG. 1

FIG. 1

(A) Northern blot analysis of erp transcripts. Isolate B31 was grown in culture medium at either a constant temperature of 23°C (labeled 23°C) or shifted from 23°C to 35°C (labeled 35°C). Filters were individually incubated with radiolabeled probes specific for the B31 gene indicated above each panel. (B) Each filter was rehybridized with a probe specific for the constitutively expressed flaB gene (7). RNA molecular size markers (in kilobases) are indicated to the left of each panel.

FIG. 2

FIG. 2

Immunoblot analysis of recombinant B31 Erp proteins. The filter was incubated with a 1:200 dilution of serum from a mouse infected with B31 by tick bite. The immunoblot signal strengths were greater from the recombinant ErpA/I and ErpB2/J proteins than from the other proteins, and the exposure time of ErpA/I and ErpB2/J in this figure was approximately 1/10 of that of the other Erp proteins. All of the recombinant Erp proteins migrated with apparent molecular masses that were greater than predicted from their sequences (Table 2). Some of the recombinant protein preparations contained probable degradation products or multimeric proteins, resulting in the presence of multiple bands. Molecular masses (in kilodaltons) are indicated to the left.

FIG. 3

FIG. 3

Identification of differentially synthesized B31 antigens. (A) Immunoblot of B31 lysates grown at a constant 23°C or shifted from 23 to 35°C. The filter was incubated with a 1:500 dilution of serum from a mouse that was infected with B31 by tick bite. (B) Immunoblots of a B31 lysate shifted from 23 to 35°C. A 1:500 dilution of pooled sera from three mice infected with B31 by tick bite was preadsorbed with each recombinant B31 Erp protein before incubation with the filter strip. Asterisks indicate the positions of bands that were absent or diminished on immunoblots prepared with sera preadsorbed with recombinant ErpA/I, ErpB2/J, or ErpK. Molecular masses (in kilodaltons) are indicated to the left of each panel.

References

    1. Akins D R, Porcella S F, Popova T G, Shevchenko D, Baker S I, Li M, Norgard M V, Radolf J D. Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue. Mol Microbiol. 1995;18:507–520. -PubMed
    1. Amouriaux P, Assous M, Margarita D, Baranton G, Saint Girons I. Polymerase chain reaction with the 30-kb circular plasmid of Borrelia burgdorferi B31 as a target for detection of the Lyme borreliosis agents in cerebrospinal fluid. Res Microbiol. 1993;144:211–219. -PubMed
    1. Barbour A G. Plasmid analysis of Borrelia burgdorferi, the Lyme disease agent. J Clin Microbiol. 1988;26:475–478. -PMC -PubMed
    1. Barbour A G, Tessier S L, Stoenner H G. Variable major proteins of Borrelia hermsii. J Exp Med. 1982;156:1312–1324. -PMC -PubMed
    1. Benach J L, Coleman J L, Skinner R A, Bosler E M. Adult Ixodes dammini on rabbits: a hypothesis for the development and transmission of Borrelia burgdorferi. J Infect Dis. 1987;155:1300–1306. -PubMed

MeSH terms

Substances

LinkOut - more resources