Nasopharyngeal commensals like Streptococcus pneumoniae, nontypeable Haemophilus influenzae (NTHi) and Moraxella catarrhalis are commonly associated with respiratory tract infections. A viral infection triggers a pro-inflammatory response leading to up-regulation of adhesion proteins and host cell surface receptors facilitating increased bacterial adherence. Microbe-host interactions closely correlate with nasopharyngeal colonisation and infection, with synergistic or competitive relationship existing. Sometimes antibiotic treatment fails with bacterial biofilms considered as etiological factors in bacterial persistence. This study investigated how polymicrobial infection and antecedent viral infection modulate cell responses to promote colonisation and suppression of the cell’s innate inflammatory response by using different respiratory cell types representing different areas of the respiratory tract. We studied the association between different strains of NTHi and S. pneumoniae and the host, and the effect of host cell contact on biofilm formation. Results showed that adenovirus infection both increased the expression of CEACAM-1, ICAM-1, and PAF-r, and also bacterial adherence, and bacterial co-infection altered the adherence in a cell type-dependent manner. Bacterial co-infection resulted in synergistic increase in the production of IL-6 and IL-8, however adenovirus infection suppressed the host cell’s pro-inflammatory response. The presence of another bacterial species and certain strains have different propensities for influencing greater biofilm formation and host-cell contact is a key contributing factor in increased biofilm formation.