title: Development of a simultaneous high resolution typing method for three SLA class II genes, SLA-DQA, SLA-DQB1, and SLA-DRB1 and the analysis of SLA class II haplotypes.
abstract: The characterization of the genetic variations of major histocompatibility complex (MHC) is essential to understand the relationship between the genetic diversity of MHC molecules and disease resistance and susceptibility in adaptive immunity. We previously reported the development of high-resolution individual locus typing methods for three of the most polymorphic swine leukocyte antigens (SLA) class II loci, namely, SLA-DQA, SLA-DQB1, and SLA-DRB1. In this study, we extensively modified our previous protocols and developed a method for the simultaneous amplification of the three SLA class II genes and subsequent analysis of individual loci using direct sequencing. The unbiased and simultaneous amplification of alleles from the all three hyper-polymorphic and pseudogene containing genes such as MHC genes is extremely challenging. However, using this method, we demonstrated the successful typing of SLA-DQA, SLA-DQB1, and SLA-DRB1 for 31 selected individuals comprising 26 different SLA class II haplotypes which were identified from 700 animals using the single locus typing methods. The results were identical to the known genotypes from the individual locus typing. The new method has significant benefits over the individual locus typing, including lower typing cost, use of less biomaterial, less effort and fewer errors in handling large samples for multiple loci. We also extensively characterized the haplotypes of SLA class II genes and reported three new haplotypes. Our results should serve as a basis to investigate the possible association between polymorphisms of MHC class II and differences in immune responses to exogenous antigens.
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, South Korea.
authors.forename: MinhThong
authors.initials: M
authors.lastname: Le
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, South Korea.
authors.forename: Hojun
authors.initials: H
authors.lastname: Choi
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, South Korea.
authors.forename: Min-Kyeung
authors.initials: MK
authors.lastname: Choi
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, South Korea.
authors.forename: Hyesun
authors.initials: H
authors.lastname: Cho
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, South Korea.
authors.forename: Jin-Hoi
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, South Korea.
authors.forename: Han Geuk
authors.initials: HG
authors.lastname: Seo
authors.affiliation: College of Veterinary Medicine, Chonbuk National University, South Korea.
authors.forename: Se-Yeon
authors.initials: SY
authors.lastname: Cha
authors.affiliation: College of Veterinary Medicine, Konkuk University, Seoul, South Korea.
authors.forename: Kunho
authors.initials: K
authors.lastname: Seo
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, South Korea. Electronic address: edenhailu@yahoo.com.
authors.forename: Hailu
authors.initials: H
authors.lastname: Dadi
authors.affiliation: Department of Animal Biotechnology, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, South Korea. Electronic address: chankyu@konkuk.ac.kr.
authors.forename: Chankyu
authors.initials: C
authors.lastname: Park
mesh_terms: D000818:Animals; D017931:DNA Primers; D059866:HLA-DQ beta-Chains / Q000235:genetics; D006239:Haplotypes; D000949:Histocompatibility Antigens Class II / Q000235:genetics*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D034421:Sus scrofa / Q000235:genetics*
chemical_list: D017931:DNA Primers; D059866:HLA-DQ beta-Chains; C057615:HLA-DQbeta antigen; D000949:Histocompatibility Antigens Class II
keywords: DQA; DQB1 and DRB1; SLA; Swine