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Infection and Immunity, September 2001, p. 5375-5384, Vol. 69, No. 9
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

Jeffrey B. Kaplan,1,* Malcolm B. Perry,2 Leann L. MacLean,2 David Furgang,1 Mark E. Wilson,1,dagger and Daniel H. Fine1

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 right-arrow2)-alpha -L-Rhap-(1-3)-2-O-(beta -D-GalpNAc)-alpha -L-Rhap-(1right-arrow. 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 IS903phi 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.

dagger Deceased.

Infection and Immunity, September 2001, p. 5375-5384, Vol. 69, No. 9
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.9.5375-5384.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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