Co-infection of H9N2 subtype avian influenza virus and infectious bronchitis virus decreases SP-A expression level in chickens.

Chicken surfactant protein A (cSP-A) is a collectin believed to play an important role in antiviral immunity. However, cSP-A expression in the respiratory tract of chickens after viral co-infection remains unclear. The aim of this study was the detection and characterization of cSP-A in co-infected chickens. For this purpose, four-week-old specific pathogen-free (SPF) chickens were divided into five groups and inoculated intranasally with H9N2 subtype avian influenza virus (AIV), infectious bronchitis virus (IBV), or Newcastle disease virus (NDV). Chickens were sacrificed at three days post inoculation, and the lung, trachea, and air sac samples were taken to determine histological changes and expression levels of cSP-A mRNA and cSP-A protein. The cSP-A mRNA and its protein were detected separately using real-time quantitative reverse transcriptional polymerase chain reaction (qRT-PCR), a sandwich enzyme-linked immunosorbent assay (S-ELISA), and an immunohistochemistry assay (IHC). In comparison, for the PBS group as the negative group and the NDV-infected group as the positive group, the histological changes showed that the lesions of the AIV+ IBV co-infected group were more serious compared to the AIV-infected group and the IBV-infected group. Consequently, the expression level of cSP-A in the AIV+IBV co-infected group significantly decreased when compared to the AIV-infected group and the IBV-infected group by qRT-PCR, ELISA, and IHC analysis. The mechanism of the downregulation of SP-A expression level will be addressed in future.