We reported earlier that antibody in sera collected from seven children with acute otitis media (AOM) due to nontypeable Haemophilus influenzae (NTHI) recognized immunodominant regions of P5-fimbrin just as we had observed in a chinchilla model of experimental NTHI-induced AOM. To expand upon those preliminary findings, we further characterized pediatric serum antibodies directed against this adhesin during AOM. Collectively, the data show that children respond immunologically to P5-fimbrin and they do so in a manner that allows for the distinction of sequence diversity within short linear peptides representing a focused region of this surface-exposed protein. The immune recognition we observed encourages us to further develop a P5-fimbrin based vaccine component.