title: Characterization of MHC class I alleles in sooty mangabeys as a tool for evaluating cellular immunity in natural hosts of SIV infection.
abstract: Although immune pressure exerted by MHC class I-restricted cytotoxic T lymphocytes (CTL) are an important determinant of outcome in pathogenic HIV and SIV infection, lack of data on MHC class I genes has hampered study of its role in natural hosts with nonpathogenic SIV infection. In this study, we cloned and characterized full-length MHC class I genes derived from the cDNA library of two unrelated naturally infected sooty mangabeys (Cercocebus atys) in whom SIV-specific CTL epitopes were previously mapped. Twenty one full-length MHC class I alleles consisting of five MHC-A (Ceat-A), 13 MHC-B (Ceat-B), and three MHC-E (Ceat-E) alleles were identified. Sequence-specific primers (SSP) for high-throughput screening of genomic DNA by PCR were developed for 16 of the 18 Ceat-A and Ceat-B alleles. Screening of 62 SIV-negative and 123 SIV-infected sooty mangabeys at the Yerkes National Primate Research Center (YNPRC) revealed the presence of up to four MHC-A and eight MHC-B alleles in individual mangabeys, indicating that similar to macaque species, mangabeys have at least two duplications of the MHC-A locus and four duplications of the MHC-B locus in the absence of an MHC-C locus. Using stable transfectants of Ceat MHC Class I alleles in the MHC-null 721.221 cell line, we identified Ceat-B*12:01 as the restricting allele of a previously reported Nef20-28 CTL epitope. Ceat-B*1201/Nef20-28 tetramers identified tetramer-positive CD8+ T lymphocytes in Ceat-B*1201-positive SIV-infected mangabeys. This study has laid the groundwork for comprehensive analysis of CTL and SIV evolution in a natural host of SIV infection.
authors.affiliation: Division of Immunology, Harvard Medical School, New England Primate Research Center, One Pine Hill Drive, Southborough, MA, 01772-9102, USA.
authors.forename: Zichun
authors.initials: Z
authors.lastname: Wang
authors.forename: Benjamin
authors.initials: B
authors.lastname: Metcalf
authors.forename: Melissa
authors.initials: M
authors.lastname: Kasheta
authors.forename: Caitlin
authors.initials: C
authors.lastname: Kasala-Hallinan
authors.forename: Dollnovan
authors.initials: D
authors.lastname: Tran
authors.forename: R Paul
authors.initials: RP
authors.lastname: Johnson
authors.forename: James G
authors.initials: JG
authors.lastname: Else
authors.forename: Julie
authors.initials: J
authors.lastname: Karl
authors.forename: David
authors.initials: D
authors.lastname: O'Connor
authors.forename: Cristian
authors.initials: C
authors.lastname: Apetrei
authors.forename: Amitinder
authors.initials: A
authors.lastname: Kaur
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D016665:Cercocebus atys / Q000235:genetics*; D018984:Epitopes, T-Lymphocyte / Q000235:genetics*; D015395:Histocompatibility Antigens Class I / Q000235:genetics*; D007111:Immunity, Cellular / Q000276:immunology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D016097:Simian Acquired Immunodeficiency Syndrome / Q000276:immunology* / Q000821:virology; D015302:Simian Immunodeficiency Virus / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
chemical_list: D018984:Epitopes, T-Lymphocyte; D015395:Histocompatibility Antigens Class I