Previous studies have demonstrated that the Moraxella catarrhalis surface antigen UspA1 is an adhesin for Chang human conjunctival cells. The present report demonstrates that lack of UspA1 expression does not affect the adherence of strain O35E to A549 human lung cells or primary cultures of human middle ear epithelial (HMEE) cells. These results imply that another molecule mediates the adherence of M. catarrhalis to these two cell lines. To identify this adhesin, strain O35E was mutagenized with a transposon and 1,000 mutants were screened in a microcolony formation assay using A549 cells. Nine independent isolates exhibited an 8-to 19-fold reduction in adherence and contained a transposon in the same locus. Nucleotide sequence data and PCR analysis indicated that the transposons were inserted in different locations in the gene encoding the surface protein Hag. Quantitative assays using one representative transposon mutant, O35E.TN2, showed considerably decreased binding to A549 as well as HMEE cells. However, this mutant adhered at wild-type levels to Chang conjunctival cells. These findings suggest that the M. catarrhalis Hag protein is an adhesin for cell lines derived from human lung and middle ear tissues.Moraxella catarrhalis is a significant pathogen of the human respiratory tract. This gram-negative bacterium is now recognized as a leading cause of otitis media (middle ear infection), together with Streptococcus pneumoniae and nontypeable isolates of Haemophilus influenzae (11,15,28,39). More than 80% of infants have at least one episode of ear infection by the age of 3 years, and M. catarrhalis causes 15 to 20% of all cases (9,14,28,29,31,39,50,59). In the United States, ϳ24 million office visits are made annually for the treatment of otitis media (29,30,39,59) and of these, roughly a sixth are caused by M. catarrhalis. This organism also causes 10 to 35% of all cases of lower respiratory tract infections in adults that are suffering from chronic obstructive pulmonary disease (39,52,53). This disease is the fourth leading cause of death in the United States, surpassed only by heart attacks, cancer, and stroke (3, 43). Lower respiratory tract infections contribute substantially to the progression of chronic obstructive pulmonary disease; thus, infectious exacerbations due to M. catarrhalis constitute an important health problem.M. catarrhalis has also been associated with diseases such as wound infections, bronchitis, conjunctivitis, sinusitis, bacteremia, pneumonia, meningitis, pericarditis, and endocarditis (8,10,11,28,39,44,55,60,63,64). Patients with underlying conditions are particularly susceptible. Long considered to be a nonpathogenic inhabitant of the human nasopharynx, M. catarrhalis has clearly emerged as an important cause of infectious diseases, and under unpredictable circumstances (e.g., immunosuppressive conditions or chronic disease), the organism can be virulent and cause serious organ complications.M. catarrhalis infections are a matter of concern due to the lack of a vaccine and the ...