Limited surface exposure of Borrelia burgdorferi outer surface lipoproteins

David L. Cox, Darrin R. Akins, Ken W. Bourell, Pekka Lahdenne, Michael V. Norgard, Justin D. Radolf

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

We used novel immunofluorescence strategies to demonstrate that outer surface proteins (Osps) A, B, and C of Borrelia burgdorferi have limited surface exposure, a finding that contradicts the prevailing viewpoint that these antigens are exclusively surface exposed. Light labeling was observed when antibodies to OspA or OspB were added to motile organisms, whereas intense fluorescence was observed when the same slides were methanol-fixed and reprobed. Modest labeling also was observed when spirochetes encapsulated in agarose beads (gel microdroplets) were incubated with antibodies to these same two antigens. This contrasted with the intense fluorescence observed when encapsulated spirochetes were probed in the presence of 0.06% Triton X- 100, which selectively removed outer membranes. Proteinase K (PK) treatment of encapsulated spirochetes abrogated surface labeling. However, PK-treated spirochetes fluoresced intensely after incubation with antibodies to OspA or OspB in the presence of detergent, confirming the existence of large amounts of subsurface Osp antigens. Modest surface labeling once again was detected when PK-treated spirochetes were reprobed after overnight incubation, a result consistent with the existence of a postulated secretory apparatus that shuttles lipoproteins to the borrelial surface. Last, experiments with the OspC-expressing B. burgdorferi strain 297 revealed that this antigen was barely detectable on spirochetal surfaces even though it was a major constituent of isolated outer membranes. We propose a model of B. burgdorferi molecular architecture that helps to explain spirochetal persistence during chronic Lyme disease.

Original languageEnglish (US)
Pages (from-to)7973-7978
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number15
DOIs
StatePublished - Jul 23 1996

Fingerprint

Spirochaetales
Borrelia burgdorferi
Lipoproteins
Endopeptidase K
Antigens
Antibodies
Fluorescence
Membranes
Lyme Disease
Octoxynol
Surface Antigens
Detergents
Sepharose
Fluorescent Antibody Technique
Methanol
Chronic Disease
Gels
Light

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Limited surface exposure of Borrelia burgdorferi outer surface lipoproteins. / Cox, David L.; Akins, Darrin R.; Bourell, Ken W.; Lahdenne, Pekka; Norgard, Michael V.; Radolf, Justin D.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 93, No. 15, 23.07.1996, p. 7973-7978.

Research output: Contribution to journalArticle

Cox, David L. ; Akins, Darrin R. ; Bourell, Ken W. ; Lahdenne, Pekka ; Norgard, Michael V. ; Radolf, Justin D. / Limited surface exposure of Borrelia burgdorferi outer surface lipoproteins. In: Proceedings of the National Academy of Sciences of the United States of America. 1996 ; Vol. 93, No. 15. pp. 7973-7978.
@article{11790fe86b0c41158d847d369e4d7f40,
title = "Limited surface exposure of Borrelia burgdorferi outer surface lipoproteins",
abstract = "We used novel immunofluorescence strategies to demonstrate that outer surface proteins (Osps) A, B, and C of Borrelia burgdorferi have limited surface exposure, a finding that contradicts the prevailing viewpoint that these antigens are exclusively surface exposed. Light labeling was observed when antibodies to OspA or OspB were added to motile organisms, whereas intense fluorescence was observed when the same slides were methanol-fixed and reprobed. Modest labeling also was observed when spirochetes encapsulated in agarose beads (gel microdroplets) were incubated with antibodies to these same two antigens. This contrasted with the intense fluorescence observed when encapsulated spirochetes were probed in the presence of 0.06{\%} Triton X- 100, which selectively removed outer membranes. Proteinase K (PK) treatment of encapsulated spirochetes abrogated surface labeling. However, PK-treated spirochetes fluoresced intensely after incubation with antibodies to OspA or OspB in the presence of detergent, confirming the existence of large amounts of subsurface Osp antigens. Modest surface labeling once again was detected when PK-treated spirochetes were reprobed after overnight incubation, a result consistent with the existence of a postulated secretory apparatus that shuttles lipoproteins to the borrelial surface. Last, experiments with the OspC-expressing B. burgdorferi strain 297 revealed that this antigen was barely detectable on spirochetal surfaces even though it was a major constituent of isolated outer membranes. We propose a model of B. burgdorferi molecular architecture that helps to explain spirochetal persistence during chronic Lyme disease.",
author = "Cox, {David L.} and Akins, {Darrin R.} and Bourell, {Ken W.} and Pekka Lahdenne and Norgard, {Michael V.} and Radolf, {Justin D.}",
year = "1996",
month = "7",
day = "23",
doi = "10.1073/pnas.93.15.7973",
language = "English (US)",
volume = "93",
pages = "7973--7978",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "15",

}

TY - JOUR

T1 - Limited surface exposure of Borrelia burgdorferi outer surface lipoproteins

AU - Cox, David L.

AU - Akins, Darrin R.

AU - Bourell, Ken W.

AU - Lahdenne, Pekka

AU - Norgard, Michael V.

AU - Radolf, Justin D.

PY - 1996/7/23

Y1 - 1996/7/23

N2 - We used novel immunofluorescence strategies to demonstrate that outer surface proteins (Osps) A, B, and C of Borrelia burgdorferi have limited surface exposure, a finding that contradicts the prevailing viewpoint that these antigens are exclusively surface exposed. Light labeling was observed when antibodies to OspA or OspB were added to motile organisms, whereas intense fluorescence was observed when the same slides were methanol-fixed and reprobed. Modest labeling also was observed when spirochetes encapsulated in agarose beads (gel microdroplets) were incubated with antibodies to these same two antigens. This contrasted with the intense fluorescence observed when encapsulated spirochetes were probed in the presence of 0.06% Triton X- 100, which selectively removed outer membranes. Proteinase K (PK) treatment of encapsulated spirochetes abrogated surface labeling. However, PK-treated spirochetes fluoresced intensely after incubation with antibodies to OspA or OspB in the presence of detergent, confirming the existence of large amounts of subsurface Osp antigens. Modest surface labeling once again was detected when PK-treated spirochetes were reprobed after overnight incubation, a result consistent with the existence of a postulated secretory apparatus that shuttles lipoproteins to the borrelial surface. Last, experiments with the OspC-expressing B. burgdorferi strain 297 revealed that this antigen was barely detectable on spirochetal surfaces even though it was a major constituent of isolated outer membranes. We propose a model of B. burgdorferi molecular architecture that helps to explain spirochetal persistence during chronic Lyme disease.

AB - We used novel immunofluorescence strategies to demonstrate that outer surface proteins (Osps) A, B, and C of Borrelia burgdorferi have limited surface exposure, a finding that contradicts the prevailing viewpoint that these antigens are exclusively surface exposed. Light labeling was observed when antibodies to OspA or OspB were added to motile organisms, whereas intense fluorescence was observed when the same slides were methanol-fixed and reprobed. Modest labeling also was observed when spirochetes encapsulated in agarose beads (gel microdroplets) were incubated with antibodies to these same two antigens. This contrasted with the intense fluorescence observed when encapsulated spirochetes were probed in the presence of 0.06% Triton X- 100, which selectively removed outer membranes. Proteinase K (PK) treatment of encapsulated spirochetes abrogated surface labeling. However, PK-treated spirochetes fluoresced intensely after incubation with antibodies to OspA or OspB in the presence of detergent, confirming the existence of large amounts of subsurface Osp antigens. Modest surface labeling once again was detected when PK-treated spirochetes were reprobed after overnight incubation, a result consistent with the existence of a postulated secretory apparatus that shuttles lipoproteins to the borrelial surface. Last, experiments with the OspC-expressing B. burgdorferi strain 297 revealed that this antigen was barely detectable on spirochetal surfaces even though it was a major constituent of isolated outer membranes. We propose a model of B. burgdorferi molecular architecture that helps to explain spirochetal persistence during chronic Lyme disease.

UR - http://www.scopus.com/inward/record.url?scp=0029840052&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029840052&partnerID=8YFLogxK

U2 - 10.1073/pnas.93.15.7973

DO - 10.1073/pnas.93.15.7973

M3 - Article

VL - 93

SP - 7973

EP - 7978

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 15

ER -