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Infection and Immunity, September 2001, p. 5375-5384, Vol. 69, No. 9
Department of Oral Pathology, Biology and
Diagnostic Sciences, New Jersey Dental School, Newark, New
Jersey,1 and Institute for
Biological Sciences, National Research Council, Ottawa, Ontario,
Canada2
Received 15 February 2001/Returned for modification 20 April
2001/Accepted 25 May 2001
The oral bacterium Actinobacillus
actinomycetemcomitans is implicated as a causative agent of
localized juvenile periodontitis (LJP). A.
actinomycetemcomitans is classified into five serotypes (a to
e) corresponding to five structurally and antigenically distinct O
polysaccharide (O-PS) components of their respective lipopolysaccharide
molecules. Serotype b has been reported to be the dominant serotype
isolated from LJP patients. We determined the lipopolysaccharide O-PS
structure from A. actinomycetemcomitans CU1000, a strain
isolated from a 13-year-old African-American female with LJP which had
previously been classified as serotype b. The O-PS of strain CU1000
consisted of a trisaccharide repeating unit composed of
L-rhamnose and 2-acetamido-2-deoxy-D-galactose (molar ratio, 2:1) with the structure
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.9.5375-5384.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Structural and Genetic Analyses of O Polysaccharide
from Actinobacillus actinomycetemcomitans Serotype
f
and
2)-
-L-Rhap-(1-3)-2-O-(
-D-GalpNAc)-
-L-Rhap-(1
. O-PS from strain CU1000 was structurally and antigenically distinct from the O-PS molecules of the five known A.
actinomycetemcomitans serotypes. Strain CU1000 was mutagenized
with transposon IS903
kan, and three
mutants that were deficient in O-PS synthesis were isolated. All three
transposon insertions mapped to a single 1-kb region on the chromosome.
The DNA sequence of a 13.1-kb region surrounding these transposon
insertions contained a cluster of 14 open reading frames that was
homologous to gene clusters responsible for the synthesis of A.
actinomycetemcomitans serotype b, c, and e O-PS antigens. The
CU1000 gene cluster contained two genes that were not present in
serotype-specific O-PS antigen clusters of the other five known
A. actinomycetemcomitans serotypes. These data indicate
that strain CU1000 should be assigned to a new A.
actinomycetemcomitans serotype, designated serotype f. A PCR
assay using serotype-specific PCR primers showed that 3 out of 20 LJP
patients surveyed (15%) harbored A.
actinomycetemcomitans strains carrying the serotype f gene
cluster. The finding of an A. actinomycetemcomitans
serotype showing serological cross-reactivity with anti-serotype
b-specific antiserum suggests that a reevaluation of strains previously
classified as serotype b may be warranted.
*
Corresponding author. Mailing address: Department of
Oral Pathology, Biology and Diagnostic Sciences, New Jersey Dental
School, MSB Room C-636, 185 S. Orange Ave., Newark, NJ 07103-2714. Phone: (973) 972-5051. Fax: (973) 972-0045. E-mail:
kaplanjb{at}umdnj.edu.
Deceased.
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