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Infection and Immunity, June 2003, p. 3419-3428, Vol. 71, No. 6
0019-9567/03/$08.00+0 DOI: 10.1128/IAI.71.6.3419-3428.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
Antigenic Composition of Borrelia burgdorferi during Infection of SCID Mice
Timothy R. Crother,1,2* Cheryl I. Champion,1,2 Xiao-Yang Wu,1,2 David R. Blanco,1,2 James N. Miller,2 and Michael A. Lovett1
Departments of Medicine,1
Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, Los Angeles, California2
Received 6 November 2002/
Returned for modification 16 December 2002/
Accepted 7 March 2003
The general concept that during infection of mice the Borrelia burgdorferi surface protein composition differs profoundly from that of tick-borne or in vitro-cultivated spirochetes is well established. Specific knowledge concerning the differences is limited because the small numbers of spirochetes present in tissue have not been amenable to direct compositional analysis. In this report we describe novel means for studying the antigenic composition of host-adapted Borrelia (HAB). The detergent Triton X-114 was used to extract the detergent-phase HAB proteins from mouse ears, ankles, knees, and hearts. Immunoblot analysis revealed a profile distinct from that of in vitro-cultivated Borrelia (IVCB). OspA and OspB were not found in the tissues of SCID mice 17 days after infection. The amounts of antigenic variation protein VlsE and the relative amounts of its transcripts were markedly increased in ear, ankle, and knee tissues but not in heart tissue. VlsE existed as isoforms having both different unit sizes and discrete lower molecular masses. The hydrophobic smaller forms of VlsE were also found in IVCB. The amounts of the surface protein (OspC) and the decorin binding protein (DbpA) were increased in ear, ankle, knee, and heart tissues, as were the relative amounts of their transcripts. Along with these findings regarding VlsE, OspC, and DbpA, two-dimensional immunoblot analysis with immune sera also revealed additional details of the antigenic composition of HAB extracted from ear, heart, and joint tissues. A variety of novel antigens, including antigens with molecular masses of 65 and 30 kDa, were found to be upregulated in mouse tissues. Extraction of hydrophobic B. burgdorferi antigens from tissue provides a powerful tool for determining the antigenic composition of HAB.
* Corresponding author. Mailing address: Department of Medicine, Division of Infectious Diseases, University of California, Los Angeles, 37-121 Center for Health Sciences, 10833 LeConte Ave., Los Angeles, CA 90095. Phone: (310) 825-4188. Fax: (310) 267-2265. E-mail: tcrother{at}ucla.edu.
Editor: D. L. Burns
Infection and Immunity, June 2003, p. 3419-3428, Vol. 71, No. 6
0019-9567/03/$08.00+0 DOI: 10.1128/IAI.71.6.3419-3428.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
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Copyright © 2003 by the American Society for Microbiology. All rights reserved.