Identification of DRB alleles in rhesus monkeys using polymerase chain reaction-sequence-specific primers (PCR-SSP) amplification

A. Lobashevsky, J. P. Smith, J. Kasten-Jolly, H. Horton, L. Knapp, R. E. Bontrop, David Watkins, Judith Thomas

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Major histocompatibility complex (MHC) class II molecules play a vital role in the regulation of T-cell functions in the mammalian immune system. Two key features characterize the polymorphism of MHC haplotypes in humans and non-human primates: the existence of a large number of alleles, and the high degree of genetic diversity between those alleles. Rhesus monkeys and Chimpanzees have been extensively used as relevant models for human diseases and transplantation. We have investigated DRB genes in 19 macaques, members of 3 families, using polymerase chain reaction with sequence-specific primers (PCR-SSP) and denaturing gradient gel electrophoresis (DGGE). After amplification PCR products were purified and subjected direct sequencing. Seven animals (Madison 1) were typed by DDGE also. We report that the DRB haplotypes defined by PCR-SSP exhibit a high degree of concordance with the data obtained by DGGE and direct sequening. Our data show prominent variability in the number of DRB1 alleles ranging from 1-4 per genotype within these families. This analysis demonstrated that most of the amplicons were identical to Mamu-DRB alleles that our PCR primers were to amplify However, 98-99% similarity was noticed in the case of Mamu-DRB1*0303, Mamu-DRB6*0103 and Mamu-DRB*W201 alleles. The observed mismatches were located in non-polymorphic regions. Thus, family studies in rhesus macaques performed by molecular methods confirmed the multiplicity of Mamu-DRB1 alleles per haplotype and the existence of allelic associations published earlier. In addition, we propose 3 more DRB allele associations (haplotypes): Mamu-DRB1*04-DRB5*03; Mamu-DRB1*04-*DRB*W5; Mamu-DRB1*04*W2. The proposed medium-resolution PCR-SSP technique appears to be a highly reproducible and discriminatory typing method for detecting polymorphisms of DRB genes in rhesus monkeys.

Original languageEnglish
Pages (from-to)254-263
Number of pages10
JournalTissue Antigens
Volume54
Issue number3
DOIs
StatePublished - Sep 30 1999
Externally publishedYes

Fingerprint

Dichlororibofuranosylbenzimidazole
Macaca mulatta
Alleles
Polymerase Chain Reaction
Haplotypes
Denaturing Gradient Gel Electrophoresis
Major Histocompatibility Complex
Pan troglodytes
Macaca
Primates
Genes
Immune System
Transplantation
Genotype
T-Lymphocytes

Keywords

  • Haplotype
  • HLA-DRB
  • PCR-SPP
  • Rhesus monkeys

ASJC Scopus subject areas

  • Immunology
  • Cell Biology

Cite this

Lobashevsky, A., Smith, J. P., Kasten-Jolly, J., Horton, H., Knapp, L., Bontrop, R. E., ... Thomas, J. (1999). Identification of DRB alleles in rhesus monkeys using polymerase chain reaction-sequence-specific primers (PCR-SSP) amplification. Tissue Antigens, 54(3), 254-263. https://doi.org/10.1034/j.1399-0039.1999.540306.x

Identification of DRB alleles in rhesus monkeys using polymerase chain reaction-sequence-specific primers (PCR-SSP) amplification. / Lobashevsky, A.; Smith, J. P.; Kasten-Jolly, J.; Horton, H.; Knapp, L.; Bontrop, R. E.; Watkins, David; Thomas, Judith.

In: Tissue Antigens, Vol. 54, No. 3, 30.09.1999, p. 254-263.

Research output: Contribution to journalArticle

Lobashevsky, A. ; Smith, J. P. ; Kasten-Jolly, J. ; Horton, H. ; Knapp, L. ; Bontrop, R. E. ; Watkins, David ; Thomas, Judith. / Identification of DRB alleles in rhesus monkeys using polymerase chain reaction-sequence-specific primers (PCR-SSP) amplification. In: Tissue Antigens. 1999 ; Vol. 54, No. 3. pp. 254-263.
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