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-DRBW1201, Caja-DRB103, Caja-DRBW16) 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-DRBW1201, 8 Caja-DRBW16 and 5 Caja-DRB103 alleles were observed. Two new DRBW16 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.