Abstract
The need for typing of the swine leukocyte antigen (SLA) is increasing with the expanded use of pigs as models for human diseases and organ-transplantation experiments, their use in infection studies, and for design of veterinary vaccines. Knowledge of SLA sequences is furthermore a prerequisite for the prediction of epitope binding in pigs. The low number of known SLA class I alleles and the limited knowledge of their prevalence in different pig breeds emphasizes the need for efficient SLA typing methods. This study utilizes an SLA class I-typing method based on next-generation sequencing of barcoded PCR amplicons. The amplicons were generated with universal primers and predicted to resolve 68–88% of all known SLA class I alleles dependent on amplicon size. We analyzed the SLA profiles of 72 pigs from four different pig populations; Göttingen minipigs and Belgian, Kenyan, and Danish fattening pigs. We identified 67 alleles, nine previously described haplotypes and 15 novel haplotypes. The highest variation in SLA class I profiles was observed in the Danish pigs and the lowest among the Göttingen minipig population, which also have the highest percentage of homozygote individuals. Highlighting the fact that there are still numerous unknown SLA class I alleles to be discovered, a total of 12 novel SLA class I alleles were identified. Overall, we present new information about known and novel alleles and haplotypes and their prevalence in the tested pig populations.
Highlights
The major histocompatibility complex class I molecules (MHC I), encoded by the highly polymorphic MHC class I genes, plays a pivotal role in the cell-mediated immune response against viral infections and cancer, and the knowledge of allele identities is crucial for prediction of vaccine responsiveness and transplantation success
Among the 72 examined pigs, we identified 50 specific alleles (16 swine leukocyte antigen (SLA)-1, 12 SLA-3, and 22 SLA-2) and 17 group-specific alleles, being alleles that could not be separated from one or two other alleles based on the sequenced part of the allele
Next-generation sequencing of amplicons generated from the transcribed exon 2 and exon 3 of the SLA class I genes is only one of several approaches for MHC class I typing
Summary
The major histocompatibility complex class I molecules (MHC I), encoded by the highly polymorphic MHC class I genes, plays a pivotal role in the cell-mediated immune response against viral infections and cancer, and the knowledge of allele identities is crucial for prediction of vaccine responsiveness and transplantation success. NGS Typing of Porcine MHC Class I (CTL’s) [1, 2]. In the entire sequence of the porcine MHC, referred to as the swine leukocyte antigen (SLA) region, between 7 and 13 classical SLA I genes (loci) spanning the centromere of chromosome, 7 have been described [3,4,5]. Despite the importance of SLA class I expression in the induction and maintenance of CTL responses, it is often not comprehensively characterized. This might be explained by the difficulties of analyzing the highly polymorphic SLA class I alleles. One very promising method for MHC class I typing is next-generation sequencing (NGS). NGS of RT-PCR-generated amplicons indicates the level of transcription for each SLA allele, as the normalized number of reads roughly correlate with the relative expression values obtained by quantitative real-time RT-PCR [7]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
