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Screening and confirmatory testing of MHC class I alleles in pig-tailed macaques

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 m... Full description

Journal Title: Immunogenetics 2011, Vol.63(8), pp.511-521
Main Author: Fernandez, Caroline
Other Authors: Reece, Jeanette , Saepuloh, Uus , Rose, Robert , Ishkandriati, Diah , O’Connor, David , Wiseman, Roger , Kent, Stephen
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 0093-7711 ; E-ISSN: 1432-1211 ; DOI: 10.1007/s00251-011-0529-5
Link: http://dx.doi.org/10.1007/s00251-011-0529-5
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recordid: springer_jour10.1007/s00251-011-0529-5
title: Screening and confirmatory testing of MHC class I alleles in pig-tailed macaques
format: Article
creator:
  • Fernandez, Caroline
  • Reece, Jeanette
  • Saepuloh, Uus
  • Rose, Robert
  • Ishkandriati, Diah
  • O’Connor, David
  • Wiseman, Roger
  • Kent, Stephen
subjects:
  • Pig-tailed macaques
  • MHC class I
  • Pyrosequencing
  • CD8+ T cell
  • Mane-A1*084:01
  • Cytotoxic T lymphocyte (CTL)
ispartof: Immunogenetics, 2011, Vol.63(8), pp.511-521
description: 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.
language: eng
source:
identifier: ISSN: 0093-7711 ; E-ISSN: 1432-1211 ; DOI: 10.1007/s00251-011-0529-5
fulltext: fulltext
issn:
  • 1432-1211
  • 14321211
  • 0093-7711
  • 00937711
url: Link


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titleScreening and confirmatory testing of MHC class I alleles in pig-tailed macaques
creatorFernandez, Caroline ; Reece, Jeanette ; Saepuloh, Uus ; Rose, Robert ; Ishkandriati, Diah ; O’Connor, David ; Wiseman, Roger ; Kent, Stephen
ispartofImmunogenetics, 2011, Vol.63(8), pp.511-521
identifier
subjectPig-tailed macaques ; MHC class I ; Pyrosequencing ; CD8+ T cell ; Mane-A1*084:01 ; Cytotoxic T lymphocyte (CTL)
descriptionPig-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.
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titleScreening and confirmatory testing of MHC class I alleles in pig-tailed macaques
descriptionPig-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.
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abstractPig-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.
copBerlin/Heidelberg
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doi10.1007/s00251-011-0529-5
pages511-521
date2011-08