Macrolide antibiotics are frequently administered to treat mycoplasmal pneumonia. However, macrolide-resistant Mycoplasma pneumoniae has recently been isolated from clinical specimens in Japan. Clarithromycin (CAM) is a 14-membered-ring macrolide that has host immunomodulatory activity. Here, we established a gnotobiotic mouse model that was monoassociated with macrolide-resistant M. pneumoniae, and pathologically and microbiologically analysed the effects of antibiotics against mycoplasmal pneumonia. We also examined the immunomodulatory activities of macrolide antibiotics in human lung carcinoma A549 cells in vitro and in a specific-pathogen-free (SPF) mouse model of pneumonia induced by M. pneumoniae antigen in vivo. CAM anti-mycoplasma antibiotics decreased the number of macrolide-sensitive and -resistant M. pneumoniae in the lungs of gnotobiotic mice. Thus, in SPF mice, CAM modulated pulmonary inflammation induced by M. pneumoniae antigens.