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1. The H9N2 subtype avian influenza virus can infect both chickens and humans. Previous studies have reported a role for erythrocytes in immunity. However, the role of H9N2 against chicken erythrocytes and the presence of complement-related genes in erythrocytes has not been studied. This research investigated the effect of H9N2 on complement-associated gene expression in chicken erythrocytes. 2. The expression of complement-associated genes ( C1s, C1q, C2, C3, C3ar1, C4, C4a, C5, C5ar1, C7, CD93  and  CFD ) was detected by reverse transcription-polymerase chain reaction (RT-PCR). Quantitative Real-Time PCR (qRT-PCR) was used to analyse the differential expression of complement-associated genes in chicken erythrocytes at 0 h, 2 h, 6 h and 10 h after the interaction between H9N2 virus and chicken erythrocytes  in vitro  and 3, 7 and 14 d after H9N2 virus nasal infection of chicks. 3. Expression levels of  C1q, C4, C1s, C2, C3, C5, C7  and  CD93  were significantly up-regulated at 2 h and significantly down-regulated at 10 h. Gene expression levels of  C1q, C3ar1, C4a, CFD and C5ar1  were seen to be different at each time point. The expression levels of  C1q, C4, C1s, C2, C3, C5, C7, CFD, C3ar1, C4a  and  C5ar1  were significantly up-regulated at 7 d and the gene expression of levels of  C3, CD93  and  C5ar1  were seen to be different at each time point. 4. The results confirmed that all the complement-associated genes were expressed in chicken erythrocytes and showed the H9N2 virus interaction with chicken erythrocytes and subsequent regulation of chicken erythrocyte complement-associated genes expression. This study reported, for the first time, the relationship between H9N2 and complement system of chicken erythrocytes, which will provide a foundation for further research into the prevention and control of H9N2 infection.

Effect of avian influenza virus subtype H9N2 on the expression of complement-associated genes in chicken erythrocytes