MHC Class II DRB genotyping is highly predictive of in-vitro alloreactivity in the common marmoset

J Immunol Methods. 2006 Jul 31;314(1-2):153-63. doi: 10.1016/j.jim.2006.06.009. Epub 2006 Jul 24.

Abstract

The common marmoset (Callithrix jacchus) is emerging as a promising alternative pre-clinical model for transplantation and immunological research. It is therefore important to establish a rapid and reliable method of confirming alloreactivity between donor-recipient pairs. In this study of a large marmoset colony (n=49), we firstly characterised MHC Class II genes (Caja-DRB*W1201, Caja-DRB1*03, Caja-DRB*W16) using, for the first time in this species, sequence-based allelic typing techniques. Exon 2 was amplified using M13-tailed PCR primers specific for known marmoset alleles, and sequenced using universal M13 sequencing primers and dye terminator cycle sequencing. Twenty-six genotypes involving monomorphic Caja-DRB*W1201, 8 Caja-DRB*W16 and 5 Caja-DRB1*03 alleles were observed. Two new DRB*W16 alleles were identified. Subsequently we investigated whether matching at MHC-DRB loci alone could accurately predict in-vitro alloreactivity as assessed by mixed lymphocyte reactions. Peripheral blood mononuclear cells (PBMC) isolated from fully and partially DRB-matched and fully mismatched animal pairs were mixed and co-cultured for T-cell proliferation. PBMC co-cultured from fully or partially mismatched pairs exhibited significant T cell proliferation above single cell controls (p<0.01). Mixed PBMC from fully DRB-matched pairs exhibited no proliferation over controls (p=0.3). Thus using Caja-DRB genotyping, suitably alloreactive donor-recipient pairs can be rapidly and accurately identified for use in further studies of cellular and solid organ transplantation.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Animals
  • Callithrix / genetics*
  • Gene Frequency
  • Genes, MHC Class II*
  • Genotype
  • Lymphocyte Culture Test, Mixed
  • Lymphocytes / immunology*
  • Molecular Sequence Data
  • Pedigree
  • Polymorphism, Genetic
  • Sequence Analysis, DNA
  • Sequence Homology, Nucleic Acid
  • Transplantation, Homologous / immunology*

Associated data

  • GENBANK/DQ340289
  • GENBANK/DQ340290