High-throughput sequencing of the porcine antibody repertoire with or without PEDV infection: A proof-of-concept study.

A host's adaptive immune system can protect against a wide variety of pathogens by producing diverse antibodies. The antibody repertoire is so vast that traditional low-throughput methods cannot fully sequence it. In this study, we developed a high-throughput sequencing (HTS) method for antibody repertoire assessment in swine, and tested it with or without porcine epidemic diarrhea virus (PEDV) infection. We isolated peripheral blood mononuclear cells from normal or PEDV-seropositive pigs and applied multiplex PCR to amplify the porcine B cell receptor heavy chain library, followed by HTS using the Illumina Miseq system to obtain full sequence information. The results from sequence analysis demonstrated that in normal conditions, several V gene segments were preferentially used, with IGHV1-4 and IGHV1S2 being the two most frequent. The IGHV usage in PEDV-seropositive pigs was not exactly the same as that of PEDV-seronegative pigs, with an increased usage of IGHV1-6. Our study provides an effective approach to comprehensively understand the overall porcine antibody repertoire, as well as to monitor broad antibody responses to viral challenge in pigs.