Identification of new cattle BoLA-DRB3 alleles by sequence-based typing

• Published in: Immunogenetics (New York) 2001-02, Vol.53 (1), p.74-81
• Main Authors: Takeshima, S, Ikegami, M, Morita, M, Nakai, Y, Aida, Y
• Format: Article
• Language: English
• Subjects: Alleles; Animals; Base Sequence; Bos; Cattle - immunology; DNA Primers; Education; Histocompatibility Antigens Class II - genetics; histocompatibility locus BoLA; Histocompatibility Testing - methods; Histocompatibility Testing - veterinary; Molecular Sequence Data; Polymerase Chain Reaction - methods; Polymorphism, Restriction Fragment Length; Sequence Alignment
• ISSN: 0093-7711; 1432-1211
• DOI: 10.1007/s002510000293

The major histocompatibility complex (MHC) of cattle is known as BoLA and is located on Chromosome 23 (Lewin 1996). The BoLA class II genes encode highly polymorphic transmembrane glycoproteins that present antigenic peptides to helper T cells and thus trigger a humoral immune response. The BoLA-DR region consists of one DRA locus and at least three DRB loci, with exon 2 of the DRB3 gene being highly polymorphic (Russell et al. 1997). BoLA-DRB3 typing has been successfully achieved using a number of methods individually or in combination (Davies et al. 1992), including serology (Davies and Antczak 1991b), mixed lymphocyte culture (Davies and Antczak 1991a), isoelectric focusing (IEF) (Glass et al. 1992; Watkins et al. 1989), restriction fragment length polymorphism (RFLP) analysis (Andersson et al. 1986), polymerase chain reaction (PCR)-RFLP (van Eijk et al. 1992), T-cell typing with cell lines and clones (Rothel et al. 1990), heteroduplex analysis (Sitte et al. 1995), sequence-specific oligonucleotide typing (Sitte et al. 1996), denaturing gradient gel electrophoresis (Aldridge et al. 1998), analysis of a microsatellite adjacent to DRB3 (Ammer et al. 1992; Ellegren et al. 1993), and sequencing of genomic DNA, cDNA, or cloned PCR products (Aida et al. 1995; Burke et al. 1991; Groenen et al. 1990; Muggli-Cockett and Stone 1989; Sigurdardottir et al. 1991; Xu et al. 1993). Most of these methods are labor intensive and their ability to define new alleles is limited, a severe drawback in studies of polymorphic loci. More precise sequence-based typing (SBT), as developed for the human MHC (HLA) (Knipper et al. 1994; Santamaria et al. 1992; Spurkland et al. 1993), would improve the resolution and accuracy of MHC allele typing in animals. In this study, we describe the development of a new method for PCR-SBT which allows identification of the BoLA-DRB3 alleles of large numbers of animals with relative ease, and we discuss the relationship between the results of PCR-SBT typing and the identification of BoLA-DRB3 alleles by PCR-RFLP analysis.