Study of MHC class II region polymorphism in the Filipino cynomolgus macaque population

Abstract

The cynomolgus macaque (Macaca fascicularis) is currently used as an animal model in various fields of immunology especially in the development of innovative vaccines for the prevention and treatment of infectious diseases. The polymorphism of the major histocompatibility complex (MHC) influences the development of adaptive immune responses and it is crucial to characterize the polymorphism of cynomolgus MHC genes. We present here a systematic study of the MHC class II haplotypes in the Filipino macaque population. By the study of a large sample of Filipino animals (N = 353), we have characterized 18 MHC class II haplotypes by means of genotyping seven microsatellites. The animals were DRB genotyped by means of PCR-SSO or DGGE-sequencing on genomic amplified fragments. We cloned and sequenced the complementary DNA (cDNA) of DQA, DQB, DPA, and DPB genes of 117 animals. Combining the microsatellite genotyping and cDNA characterized in the 117 animals, we defined genetic association between the cDNA and the microsatellites and characterized 18 MHC class II haplotypes. For 104 animals out of the 353 studied, the presence of a recombinant haplotype was highly probable. Thirty-four percent of recombination was located in 256 kb segment between D6S2876 and D6S2747 microsatellites, a region encompassing several hot spots of recombination in the human MHC.

Survey of major histocompatibility complex class II diversity in pig-tailed macaques

“…2). This is again consistent with a higher DQ region diversity for pig-tailed macaques compared to Mauritian and Filipino cynomolgus macaques, in which seven and 13 haplotypes have been described, respectively (Blancher et al 2014; O’Connor et al 2007; Wiseman et al 2013). However, many of the distinct haplotypes in the DQ region are closely-related variants of each other.…”

Discovery of novel MHC-class I alleles and haplotypes in Filipino cynomolgus macaques (Macaca fascicularis) by pyrosequencing and Sanger sequencing

“…Nevertheless, this suggests that it is possible to detect MHC homozygotes in the Filipino macaque population as well as the Mauritius population, which is the preferred population group for recruitment of animals into biomedical studies. In the class II region, the #7 and #11 are highly frequent haplotypes in Filipino macaques with 30.6 and 13.9 %, respectively (Blancher et al 2014). This suggests that MHC homozygous animals might be easily detected by large scale screening by focusing on F/A/E/B-Hp1, F/A/E/BHp2, #7 and #11 in the Filipino macaque population.…”

“…Because the results of biomedical experiments strongly depend on the immunogenetic background of animals conditioned by various environmental selective factors such as pathogens, MHC homozygous macaques are preferred for use in biomedical research (Vallender 2014 at the population level. In this regard, one of our Filipino animals (ID152) was strictly a Bnear-homozygote^that has the F/A/E/B-Hp1 in the Mafa-class I region and the #7 haplotype in the Mafa-class II region (Blancher et al 2014) on both chromosomes (tentatively named BHT1^) (Fig. 4).…”

“…The Mafa-DR-Mafa-DQ-Mafa-DP haplotypes of these animals have been previously characterized Blancher et al 2014). We also used the genomic DNA from the white blood cells of 20 unrelated cynomolgus macaques from Vietnam and Indonesia (ten from Nafovanny and ten from CV Universal Fauna Breeder and Exporter of Non-human Primates for Laboratories) for genotyping of a novel MHC class I gene, Mafa-A8*01:01.…”

“…The Mauritian and Filipino macaque populations are believed to have originated from a colonization of Indonesian/continental animals followed by a bottleneck approximately 110,000 years BP (Blancher et al 2008). In fact, the Mauritian and Filipino macaques show a relatively high genetic diversity, but a smaller number of MHC class I and class II alleles than the Indonesian and Vietnamese populations Blancher et al 2014;Blancher et al 2006;Kita et al 2009;Krebs et al 2005;Leuchte et al 2004;Sano et al 2006). Although the seven most frequent Mafa-class I haplotypes were characterized from 67 distinct Mafa-class I transcripts in the Mauritian population (Budde et al 2010), detailed polymorphic information for the Mafa-class I genes is still lacking in comparison to the Mafa-class II genes (Blancher et al 2014) in the Filipino population.…”