Quy Van Chanh Le1†, Thong Minh Le1†, Hye-Sun Cho1, Won-Il Kim2, Kwonho Hong1, Hyuk Song1, Jin-Hoi Kim1 and Chankyu Park1*
1 Department of Stem Cell and Regenerative Biotechnology, Konkuk University, Gwangjin-gu, Seoul, South Korea
2 College of Veterinary Medicine, Chonbuk National University, Iksan, Republic of Korea.
† Quy Van Chanh Le and Thong Minh Le contributed equally to this work
*Correspondence : Chankyu Park
Primary porcine alveolar macrophages (PAM) are useful for studying viral infections and immune response in pigs; however, long-term use of these cells is limited by the cells’ short lifespan. We immortalized primary PAMs by transfecting them with both hTERT and SV40LT and established two immortalized cell lines (iPAMs) actively proliferating even after 35 passages. These cells possessed the characteristics of primary PAMs, including strong expression of swine leukocyte antigen (SLA) class II genes and the inability to grow anchorage-independently. We characterized their SLA genes and subsequently performed peptide-SLA binding assays using a peptide from porcine circovirus type 2 open reading frame 2 to experimentally measure the binding affinity of the peptide to SLA class II. The number of peptides bound to cells measured by fluorescence was very low for PK15 cells (7.0%±1.5), which are not antigen-presenting cells, unlike iPAM61 (33.7%±3.4; SLA-DQA*0201/0303, DQB1*0201/0901, DRB1*0201/1301) and iPAM303 (73.3%±5.4; SLA DQA*0106/0201, DQB1*0202/0701, DRB1*0402/0602). The difference in peptide binding between the two iPAMs was likely due to the allelic differences between the SLA class II molecules that were expressed. The development of an immortal PAM cell panel harboring diverse SLA haplotypes and the use of an established method in this study can become a valuable tool for evaluating the interaction between antigenic peptides and SLA molecules and is important for many applications in veterinary medicine including vaccine development.