Discovery of novel MHC-class I alleles and haplotypes in Filipino cynomolgus macaques (Macaca fascicularis) by pyrosequencing and Sanger sequencing
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Although the low polymorphism of the major histocompatibility complex (MHC) transplantation genes in the Filipino cynomolgus macaque (Macaca fascicularis) is expected to have important implications in the selection and breeding of animals for medical research, detailed polymorphism information is still lacking for many of the duplicated class I genes. To better elucidate the degree and types of MHC polymorphisms and haplotypes in the Filipino macaque population, we genotyped 127 unrelated animals by the Sanger sequencing method and high-resolution pyrosequencing and identified 112 different alleles, 28 at cynomolgus macaque MHC (Mafa)-A, 54 at Mafa-B, 12 at Mafa-I, 11 at Mafa-E, and seven at Mafa-F alleles, of which 56 were newly described. Of them, the newly discovered Mafa-A8*01:01 lineage allele had low nucleotide similarities (<86 %) with primate MHC class I genes, and it was also conserved in the Vietnamese and Indonesian populations. In addition, haplotype estimations revealed 17 Mafa-A, 23 Mafa-B, and 12 Mafa-E haplotypes integrated with 84 Mafa-class I haplotypes and Mafa-F alleles. Of these, the two Mafa-class I haplotypes, F/A/E/B-Hp1 and F/A/E/B-Hp2, had the highest haplotype frequencies at 10.6 and 10.2 %, respectively. This suggests that large scale genetic screening of the Filipino macaque population would identify these and other high-frequency Mafa-class I haplotypes that could be used as MHC control animals for the benefit of biomedical research.
KeywordsCynomolgus macaque Major histocompatibility complex MHC Polymorphism Haplotype Next generation sequencing NGS Sanger sequencing
We are grateful to Dr. Atsushi Toyoda at Center for Information Biology, Comparative Genomics Laboratory, National Institute of Genetics for making sequence data and Mr. Masayuki Tanaka, Mr. Hideki Hayashi, and Mr. Tadayuki Sato at the Education and Research Support Center, Research and Development Division, Tokai University for their technical support. This work was supported by MEXT KAKENHI (no. 221S0002), JSPS KAKENHI (no. 21300155), and was partially supported by the program “Research Center Network for Realization of Regenerative Medicine” in Japan Agency for Medical Research and Development (AMED) and from Adaptable and Seamless Technology transfer Program in AMED.
Conflict of interest
The authors declare that they have no conflict of interest.
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