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Published on: December 23, 2016
Identification of novel MHC class I sequences in pig-tailed macaques by amplicon pyrosequencing and full-length cDNA
Claire E O'Leary1, Roger W Wiseman, Julie A Karl
1Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI 53711, USA.
Abstract:
Pig-tailed macaques (Macaca nemestrina) provide important animal models in biomedical research, but utility of this species for HIV and other disease pathogenesis research is limited by incomplete knowledge of major histocompatibility complex (MHC) class I genetics. Here, we describe comprehensive MHC class I genotyping of 24 pig-tailed macaques, using pyrosequencing to evaluate a 367- bp complementary DNA (cDNA)-PCR amplicon spanning the highly polymorphic peptide-binding region of MHC class I transcripts. We detected 29 previously described Mane transcripts, 90 novel class I sequences, and eight shared MHC class IB haplotypes. We used this genotyping data to inform full-length MHC class I cDNA allele discovery, characterizing 66 novel full-length transcripts. These new full-length sequences nearly triple the number of Mane-B cDNA sequences previously characterized. The comprehensive genotypes and full-length Mane transcripts described herein add value to pig-tailed macaques as model organisms in biomedical research; furthermore, the coordinated method for MHC genotyping and allele discovery is extensible to other less well-characterized nonhuman primate species.
Insights
This study enhances understanding of pig-tailed macaque (Macaca nemestrina) genetics, crucial for disease research. New major histocompatibility complex (MHC) class I sequences significantly expand knowledge of this important biomedical model.
Area of Science:
- Immunogenetics
- Primate genomics
- Biomedical research models
Background:
- Pig-tailed macaques (Macaca nemestrina) are vital animal models.
- Limited knowledge of their major histocompatibility complex (MHC) class I genetics hinders disease research, including HIV.
- Comprehensive genetic characterization is needed to maximize their utility.
Purpose of the Study:
- To perform comprehensive MHC class I genotyping in pig-tailed macaques.
- To discover novel full-length MHC class I cDNA alleles.
- To enhance the value of pig-tailed macaques as biomedical research models.
Main Methods:
- Pyrosequencing of a 367-bp cDNA-PCR amplicon covering the MHC class I peptide-binding region.
- Genotyping of 24 pig-tailed macaques.
- Informing full-length allele discovery based on genotyping data.
Main Results:
- Detected 29 known and 90 novel MHC class I sequences.
- Identified eight shared MHC class IB haplotypes.
- Characterized 66 novel full-length MHC class I cDNA transcripts, nearly tripling known Mane-B sequences.
Conclusions:
- This study significantly expands the characterized MHC class I repertoire in pig-tailed macaques.
- The findings enhance the utility of these macaques for disease pathogenesis research.
- The developed method is applicable to other nonhuman primate species.
