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Infection and Immunity, August 1999, p. 4153-4160, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Opacity-Associated Protein A Contributes to the Binding of Haemophilus influenzae to Chang Epithelial Cells

Nemani V. Prasadarao,1,* Elena Lysenko,2 Carol A. Wass,1 Kwang Sik Kim,1 and Jeffrey N. Weiser2

Division of Infectious Diseases, Childrens Hospital Los Angeles, and University of Southern California School of Medicine, Los Angeles, California 90027,1 and Departments of Pediatrics and Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 191042

Received 23 February 1999/Returned for modification 15 April 1999/Accepted 28 May 1999

Opacity-associated protein A (OapA), which is responsible for the transparent-colony phenotype of Haemophilus influenzae, has been implicated in the colonization of the nasopharynx in an infant rat model of carriage. In this report, we show that OapA mediates attachment to Chang epithelial cells examined by using genetically defined type b and nontypeable H. influenzae strains with or without OapA. We also showed that OapA was conserved among H. influenzae strains by comparing deduced amino acid sequences. Both recombinant OapA and polyclonal anti-OapA antiserum blocked the binding of H. influenzae to Chang epithelial cells, suggesting that the interaction of H. influenzae is specific to OapA. Moreover, the binding of recombinant OapA to epithelial cells further provided evidence that OapA can promote attachment of H. influenzae. Expression of oapA gene in a nonadherent Escherichia coli strain significantly increased the binding to Chang epithelial cells, and disruption of the oapA gene with kanamycin resistance cassette insertion resulted in a significant loss of binding. These findings demonstrate that OapA plays a role in H. influenzae binding to human conjunctival epithelial cells.

* Corresponding author. Mailing address: Division of Infectious Diseases, #51, Childrens Hospital Los Angeles, 4650 Sunset Blvd., Los Angeles, CA 90027. Phone: (323) 669-5622. Fax: (323) 660-2661. E-mail: nemani{at}hsc.usc.edu.

Infection and Immunity, August 1999, p. 4153-4160, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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