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Infect Immun, August 1998, p. 3656-3665, Vol. 66, No. 8
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Cloning and Expression of the Moraxella catarrhalis Lactoferrin Receptor Genes

Run-Pan Du,1 Qijun Wang,1 Yan-Ping Yang,1 Anthony B. Schryvers,2 Pele Chong,1 Michel H. Klein,1 and Sheena M. Loosmore1 *

Pasteur Merieux Connaught Canada Research Centre, North York, Ontario, Canada M2R 3T4,1 and Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada T2N 4N12

Received 24 February 1998/Returned for modification 31 March 1998/Accepted 1 June 1998

The lactoferrin receptor genes from two strains of Moraxella catarrhalis have been cloned and sequenced. The lfr genes are arranged as lbpB followed by lbpA, a gene arrangement found in lactoferrin and transferrin receptor operons from several bacterial species. In addition, a third open reading frame, orf3, is located one nucleotide downstream of lbpA. The deduced lactoferrin binding protein A (LbpA) sequences from the two strains were found to be 99% identical, the LbpB sequences were 92% identical, and the ORF3 proteins were 98% identical. The lbpB gene was PCR amplified and sequenced from a third strain of M. catarrhalis, and the encoded protein was found to be 77% identical and 84% similar to the other LbpB proteins. Recombinant LbpA and LbpB proteins were expressed from Escherichia coli, and antisera raised to the purified proteins were used to assess antigenic conservation in a panel of M. catarrhalis strains. The recombinant proteins were tested for the ability to bind human lactoferrin following gel electrophoresis and electroblotting, and rLbpB, but not rLbpA, was found to bind lactoferrin. Bactericidal antibody activity was measured, and while the anti-rLbpA antiserum was not bactericidal, the anti-rLbpB antisera were found to be weakly bactericidal. Thus, LbpB may have potential as a vaccine candidate.

* Corresponding author. Mailing address: Pasteur Merieux Connaught Canada Research Centre, 1755 Steeles Ave. W., North York, Ontario, Canada M2R 3T4. Phone: (416) 667-2932. Fax: (416) 667-2740. E-mail: sloosmore{at}ca.pmc-vacc.com.

Infect Immun, August 1998, p. 3656-3665, Vol. 66, No. 8
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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