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Infection and Immunity, November 2001, p. 7083-7090, Vol. 69, No. 11
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.11.7083-7090.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Analysis of Borrelia burgdorferi
vlsE Gene Expression and Recombination in the Tick
Vector
Karl J.
Indest,1,
Jerrilyn K.
Howell,2
Mary B.
Jacobs,1
Dorothy
Scholl-Meeker,1
Steven J.
Norris,2 and
Mario T.
Philipp1,*
Department of Parasitology, Tulane Regional
Primate Research Center, Tulane University Health Sciences Center,
Covington, Louisiana 70433,1 and
Department of Pathology and Laboratory Medicine and Department
of Microbiology and Molecular Genetics, University of Texas Medical
School at Houston, Houston, Texas 770302
Received 20 April 2001/Returned for modification 10 July
2001/Accepted 27 July 2001
Expression and recombination of the antigenic variation
vlsE gene of the Lyme disease spirochete Borrelia
burgdorferi were analyzed in the tick vector. To assess
vlsE expression, Ixodes scapularis nymphs
infected with the B. burgdorferi strain B31 were fed on
mice for 48 or 96 h or to repletion and then crushed and acetone
fixed either immediately thereafter (ticks collected at the two earlier
time points) or 4 days after repletion. Unfed nymphs also were
examined. At all of the time points investigated, spirochetes were able
to bind a rabbit antibody raised against the conserved invariable
region 6 of VlsE, as assessed by indirect immunofluorescence, but not
preimmune serum from the same rabbit. This same antibody also bound to
B31 spirochetes cultivated in vitro. Intensity of fluorescence appeared
highest in cultured spirochetes, followed by spirochetes present in
unfed ticks. Only a dim fluorescent signal was observed on spirochetes
at the 48 and 96 h time points and at day 4 postrepletion.
Expression of vlsE in vitro was affected by a rise in pH
from 7.0 to 8.0 at 34°C. Hence, vlsE expression
appears to be sensitive to environmental cues of the type found in the
B. burgdorferi natural history. To assess
vlsE recombination, nymphs were capillary fed the
B. burgdorferi B31 clonal isolate 5A3. Ticks thus
infected were either left to rest for 4 weeks (Group I) or fed to
repletion on a mouse (Group II). The contents of each tick from both
groups were cultured and 10 B. burgdorferi clones from
the spirochetal isolate of each tick were obtained. The
vlsE cassettes from several of these clones were
amplified by PCR and sequenced. Regardless of whether the isolate was
derived from Group I or Group II ticks, no changes were observed in the
vlsE sequence. In contrast, vlsE
cassettes amplified from B. burgdorferi clones derived
from a mouse that was infected with B31-5A3 capillary-fed nymphs showed
considerable recombination. It follows that vlsE
recombination does not occur in the tick vector.
*
Corresponding author. Mailing address: Tulane Regional
Primate Research Center, Tulane University Health Sciences Center, 18703 Three Rivers Rd., Covington, LA 70433. Phone: (504) 871-6221. Fax: (504) 871-6390. E-mail: philipp{at}tpc.tulane.edu.

Present address: ASI Services, Vicksburg, MS
39182.
Infection and Immunity, November 2001, p. 7083-7090, Vol. 69, No. 11
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.11.7083-7090.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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