Respiratory syncytial virus (RSV), a major causative agent of respiratory tract infections, influences allergic diseases. Mast cells, important effector cells in allergic disease, also express chemokine (C-X(3)-C motif) receptor 1 (CX(3)CR1). The RSV attachment glycoprotein (G protein) is structurally similar to CX(3)C ligand 1 (CX(3)CL1), the CX(3)CR1 ligand, suggesting that RSV directly interacts with and affects mast cell function, including degranulation. In this paper, the effect of RSV infection on mast cell function was studied using the human mast cell line (HMC-1). The results showed that RSV infection and replication was inefficient in HMC-1 cells than in human epithelial A549 cells. Additionally, HMC-1 degranulation occurred only in coculture with RSV-infected A549 cells, with up-regulation of TNFalpha secretion. However, direct RSV inoculation and incubation with RSV-infected A549 cell culture medium failed to induce HMC-1 degranulation, suggesting that virus-infected cells are critical for degranulation during RSV infection; however, degranulation does not occur by direct RSV infection into mast cells.