Adenoviral (Ad) vectors are currently being developed for immunotherapy and vaccine delivery, particularly for cancer and antiviral treatment (e.g., human immunodeficiency virus vaccine). However, the inefficient transduction of antigen-presenting cells of myeloid lineage such as dendritic cells (DCs) by conventional Ad5-based vectors limits these applications. DCs lack the coxsackie and adenovirus receptor but express CD46, a member of the family of complement regulatory proteins, as well as sialic acid glycoconjugates, a situation that allows infection by Ad type 37 as well as most subgroup B Ads. In this study, we generated recombinant Ad fiber knobs (FKs) to probe Ad37 receptor usage on immune cells. Remarkably, treatment of human myeloid cells with Ad37 FK (37FK) enhanced, rather than inhibited transduction by Ad5 or Ad37 pseudotyped virions. The enhanced gene transfer was dose dependent, involved association with alpha(2,6)-linked sialic acid residues, and was limited to blood cells of myeloid lineage, including immature and mature DCs. We also provide evidence that 37FK binds directly to Ad5 virus particles, likely acting as a bridge to facilitate greater virus-cell interaction.