title: Screening and confirmatory testing of MHC class I alleles in pig-tailed macaques.
abstract: Pig-tailed macaques (Macaca nemestrina) are a commonly studied primate model of human AIDS. The Mane-A1*084:01 MHC class I allele (previously named Mane-A*10) is important for the control of SIV infection by CD8+ T cells in this model. Validated methods to detect this allele in large numbers of macaques are lacking. We studied this MHC allele using sequence-specific PCRs in 217 pig-tailed macaques and identified 75 (35%) positive animals. We then performed massively parallel pyrosequencing with a universal 568-bp MHC class I cDNA-PCR amplicon for 50 of these 75 macaques. All 50 animals expressed Mane-A1*084:01 or closely related variants of the Mane-A1*084 lineage. Mane-A1*084 transcripts accounted for an average of 20.9% of all class I sequences identified per animal. SIV infection of a subset of these macaques resulted in the induction of SIV-specific CD8+ T cell responses detected by Mane-A1*084:01 tetramers. An average of 19 distinct class I transcripts were identified per animal by pyrosequencing. This analysis revealed 89 new Mane class I sequences as well as 32 previously described sequences that were extended with the longer amplicons employed in the current study. In addition, multiple Mane class I haplotypes that had been inferred previously based on shared transcript profiles between unrelated animals were confirmed for a subset of animals where pedigree information was available. We conclude that sequence-specific PCR is useful to screen pig-tailed macaques for Mane-A1*084:01, although pyrosequencing permits a much broader identification of the repertoire of MHC class I sequences and haplotypes expressed by individual animals.
authors.affiliation: Department of Microbiology and Immunology, University of Melbourne, Parkville, Vic, Australia.
authors.forename: Caroline S
authors.initials: CS
authors.lastname: Fernandez
authors.forename: Jeanette C
authors.initials: JC
authors.lastname: Reece
authors.forename: Uus
authors.initials: U
authors.lastname: Saepuloh
authors.forename: Robert
authors.initials: R
authors.lastname: De Rose
authors.forename: Diah
authors.initials: D
authors.lastname: Ishkandriati
authors.forename: David H
authors.initials: DH
authors.lastname: O'Connor
authors.forename: Roger W
authors.initials: RW
authors.lastname: Wiseman
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Kent
mesh_terms: D000483:Alleles; D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D005805:Genes, MHC Class I; D005838:Genotype; D008254:Macaca nemestrina / Q000235:genetics* / Q000276:immunology