title: MHC Class II DRB genotyping is highly predictive of in-vitro alloreactivity in the common marmoset.
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.
authors.affiliation: Transplantation Immunology Laboratory and Department of Medicine, University of Adelaide, The Queen Elizabeth Hospital Campus, 28 Woodville Road, Woodville, SA 5011, Australia.
authors.forename: Shilpanjali
authors.initials: S
authors.lastname: Prasad
authors.forename: Ian
authors.initials: I
authors.lastname: Humphreys
authors.forename: Svjetlana
authors.initials: S
authors.lastname: Kireta
authors.forename: Robert Bruce
authors.initials: RB
authors.lastname: Gilchrist
authors.forename: Peter
authors.initials: P
authors.lastname: Bardy
authors.forename: Graeme R
authors.initials: GR
authors.lastname: Russ
authors.forename: Patrick Toby Hewlett
authors.initials: PT
authors.lastname: Coates
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D002144:Callithrix / Q000235:genetics*; D005787:Gene Frequency; D005802:Genes, MHC Class II; D005838:Genotype; D007959:Lymphocyte Culture Test, Mixed; D008214:Lymphocytes / Q000276:immunology*; D008969:Molecular Sequence Data; D010375:Pedigree; D011110:Polymorphism, Genetic; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D014184:Transplantation, Homologous / Q000276:immunology*