Potential role of TRAF2 in pulmonary hypertension in broiler chickens and preparation and specificity analysis of its polyclonal antibody

Due to the lack of specific antibody anti-chicken tumor necrosis factor receptor-associated factor 2 (TRAF2), it is difficult to further explore the role of TRAF2 in pulmonary artery remodeling in pulmonary hypertension(PH) in broilers. In this experiment, we prepared a polyclonal antibody to TRAF2 by constructing a TRAF2 recombinant protein prokaryotic expression vector and analyzed the expression of TRAF2 in in vivo and in vitro models of pulmonary hypertension in broiler chickens and the effect of TRAF2 on the activity and apoptosis of PASMCs. The results showed that after immunization with TRAF2 recombinant protein we obtained high titers of polyclonal antibodies, and astragalus polysaccharide as an immune adjuvant could enhance the effect of immunization. Antibody specificity showed that the TRAF2 polyclonal antibody specifically bound to TRAF2 protein in chickens and ducks but weakly to TRAF2 protein in rabbits, mice and goats.TRAF2 was significantly upregulated in an in vivo and in vitro model of PH in broilers. Knockdown of TRAF2 inhibited the activity of PASMCs and induced apoptosis in PASMCs. Our study lays the foundation for further research on the pathomechanism of PH in broiler chickens and provides new targets for its prevention and drug development.