Abstract
Borrelia burgdorferi produces extracellular vesicles containing various borrelial protein antigens when propagated in vitro in culture media. Commonly observed components of borrelial vesicle preparations are borrelial surface antigens, bovine serum albumin, and the heavy chains of rabbit immunoglobulin G and immunoglobulin M. This study employed ultracentrifugation to harvest borrelial vesicles and analyzed these preparations by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western immunoblotting. We demonstrated that the rabbit mu heavy-chain band observed was devoid of OspA or at most levels below those detectable by immunoblot. We also demonstrated the recovery of borrelial vesicles at relative centrifugal forces as low as 25,000 x g, compared with the force of > 200,000 x g normally employed. Further, the mu heavy-chain band was recovered from uninoculated growth media processed at 25,000 x g, suggesting that it behaves as a particle rather than as a soluble molecule under these conditions. Lastly, vesicles were demonstrated to be present in preparations harvested from growth media supplemented with fetal calf serum, suggesting that vesicle production by B. burgdorferi can occur in the absence of immunoglobulins.
Full Text
The Full Text of this article is available as a PDF (459.1 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Barbour A. G., Hayes S. F. Biology of Borrelia species. Microbiol Rev. 1986 Dec;50(4):381–400. doi: 10.1128/mr.50.4.381-400.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barbour A. G., Tessier S. L., Todd W. J. Lyme disease spirochetes and ixodid tick spirochetes share a common surface antigenic determinant defined by a monoclonal antibody. Infect Immun. 1983 Aug;41(2):795–804. doi: 10.1128/iai.41.2.795-804.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Comstock L. E., Fikrig E., Shoberg R. J., Flavell R. A., Thomas D. D. A monoclonal antibody to OspA inhibits association of Borrelia burgdorferi with human endothelial cells. Infect Immun. 1993 Feb;61(2):423–431. doi: 10.1128/iai.61.2.423-431.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dorward D. W., Huguenel E. D., Davis G., Garon C. F. Interactions between extracellular Borrelia burgdorferi proteins and non-Borrelia-directed immunoglobulin M antibodies. Infect Immun. 1992 Mar;60(3):838–844. doi: 10.1128/iai.60.3.838-844.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dorward D. W., Schwan T. G., Garon C. F. Immune capture and detection of Borrelia burgdorferi antigens in urine, blood, or tissues from infected ticks, mice, dogs, and humans. J Clin Microbiol. 1991 Jun;29(6):1162–1170. doi: 10.1128/jcm.29.6.1162-1170.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garon C. F., Dorward D. W., Corwin M. D. Structural features of Borrelia burgdorferi--the Lyme disease spirochete: silver staining for nucleic acids. Scanning Microsc Suppl. 1989;3:109–115. [PubMed] [Google Scholar]
- Grenier D., Mayrand D. Functional characterization of extracellular vesicles produced by Bacteroides gingivalis. Infect Immun. 1987 Jan;55(1):111–117. doi: 10.1128/iai.55.1.111-117.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Judd R. C. 125I-peptide mapping of protein III isolated from four strains of Neisseria gonorrhoeae. Infect Immun. 1982 Aug;37(2):622–631. doi: 10.1128/iai.37.2.622-631.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Katona L. I., Beck G., Habicht G. S. Purification and immunological characterization of a major low-molecular-weight lipoprotein from Borrelia burgdorferi. Infect Immun. 1992 Dec;60(12):4995–5003. doi: 10.1128/iai.60.12.4995-5003.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lefebvre R. B., Perng G. C., Johnson R. C. The 83-kilodalton antigen of Borrelia burgdorferi which stimulates immunoglobulin M (IgM) and IgG responses in infected hosts is expressed by a chromosomal gene. J Clin Microbiol. 1990 Jul;28(7):1673–1675. doi: 10.1128/jcm.28.7.1673-1675.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Radolf J. D., Bourell K. W., Akins D. R., Brusca J. S., Norgard M. V. Analysis of Borrelia burgdorferi membrane architecture by freeze-fracture electron microscopy. J Bacteriol. 1994 Jan;176(1):21–31. doi: 10.1128/jb.176.1.21-31.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shoberg R. J., Thomas D. D. Specific adherence of Borrelia burgdorferi extracellular vesicles to human endothelial cells in culture. Infect Immun. 1993 Sep;61(9):3892–3900. doi: 10.1128/iai.61.9.3892-3900.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steere A. C., Grodzicki R. L., Kornblatt A. N., Craft J. E., Barbour A. G., Burgdorfer W., Schmid G. P., Johnson E., Malawista S. E. The spirochetal etiology of Lyme disease. N Engl J Med. 1983 Mar 31;308(13):733–740. doi: 10.1056/NEJM198303313081301. [DOI] [PubMed] [Google Scholar]
- Tai K. F., Ma Y., Weis J. J. Normal human B lymphocytes and mononuclear cells respond to the mitogenic and cytokine-stimulatory activities of Borrelia burgdorferi and its lipoprotein OspA. Infect Immun. 1994 Feb;62(2):520–528. doi: 10.1128/iai.62.2.520-528.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whitmire W. M., Garon C. F. Specific and nonspecific responses of murine B cells to membrane blebs of Borrelia burgdorferi. Infect Immun. 1993 Apr;61(4):1460–1467. doi: 10.1128/iai.61.4.1460-1467.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]