Respiratory diseases are responsible for major economic losses in poultry farms. Newcastle disease virus (NDV) infections cause huge economic losses in poultry industry especially in the presence of other co-infecting pathogens. The purpose of this study was to assess the less understood effect of Mycoplasma gallisepticum (MG) on the replication and immune responses of NDV in broiler chicken. Three-week-old commercial broiler chickens were inoculated with either NDV, MG or both etiological agents. The experimental groups were identified as follows: negative control (Group C), Mycoplasma challenged (Group M), NDV challenged (Group V) and virus and Mycoplasma challenged (Group V+M). Blood samples and swabs were collected on daily basis for two weeks. All infected birds showed positive results for NDV shedding, however, the pattern of virus shedding was different, with birds of the group V+M showing more pronounced virus shedding than the birds in the group V. In addition, birds of V+M group showed significant reduction in anti-AI antibody responses and interferon gene expression than the birds in the V group. The present study revealed that MG could facilitate replication of NDV by bringing alterations in immune responses.