Karl Julie A, Heimbruch Katelyn E, Vriezen Claire E, Mironczuk Cassandra J, Dudley Dawn M, Wiseman Roger W, O'Connor David H
Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, 53715, USA.
Immunogenetics. 2014 Nov;66(11):613-23. doi: 10.1007/s00251-014-0797-y. Epub 2014 Aug 17.
Pig-tailed macaques (Macaca nemestrina) serve as important models for human infectious disease research. Major histocompatibility complex (MHC) class II molecules are important to this research since they present peptides to CD4+ T cells. Despite the importance of characterizing the MHC-II alleles expressed in model species like pig-tailed macaques, to date, less than 150 MHC-II alleles have been named for the six most common classical class II loci (DRA, DRB, DQA, DQB, DPA, and DPB) in this population. Additionally, only a small percentage of these alleles are full-length, making it impossible to use the known sequence for reagent development. To address this, we developed a fast, high-throughput method to discover full-length MHC-II alleles and used it to characterize alleles in 32 pig-tailed macaques. By this method, we identified 128 total alleles across all six loci. We also performed an exon 2-based genotyping assay to validate the full-length sequencing results; this genotyping assay could be optimized for use in determining MHC-II allele frequencies in large cohorts of pig-tailed macaques.
食蟹猴（Macaca nemestrina）是人类传染病研究的重要模型。主要组织相容性复合体（MHC）II类分子对该研究很重要，因为它们将肽呈递给CD4+ T细胞。尽管表征在食蟹猴等模型物种中表达的MHC-II等位基因很重要，但迄今为止，该群体中六个最常见的经典II类基因座（DRA、DRB、DQA、DQB、DPA和DPB）命名的MHC-II等位基因不到150个。此外，这些等位基因中只有一小部分是全长的，这使得无法使用已知序列进行试剂开发。为了解决这个问题，我们开发了一种快速、高通量的方法来发现全长MHC-II等位基因，并使用它来表征32只食蟹猴的等位基因。通过这种方法，我们在所有六个基因座上共鉴定出128个等位基因。我们还进行了基于外显子2的基因分型检测，以验证全长测序结果；这种基因分型检测可以进行优化，用于确定大型食蟹猴群体中的MHC-II等位基因频率。
