The common marmoset as a novel preclinical transplant model: identification of new MHC class II DRB alleles and prediction of in vitro alloreactivity

Abstract

The difficulties with using nonhuman primate species such as rhesus macaques and baboons have led us to investigate the common marmoset (Callithrix jacchus) as an alternative preclinical model for transplantation research. This requires reliable methods of detecting alloreactivity between donor and recipient pairs, particularly if colonies are inbred and share just a few common alleles for leucocyte antigens. We firstly identified marmoset major histocompatibility complex (MHC) Class II DRB genes (Caja-DRB*W1201, Caja-DRB1*03, Caja-DRB*W16) using sequence-based typing techniques. Genomic DNA (n= 49) was extracted from whole blood or spleen tissue. Exon 2 of target genes was amplified by PCR using primers specific for known marmoset alleles, and then sequenced using ABI PRISM((R)) Big Dye Terminator technology and Assign sequence analysis software. DRB*W1201 was universally present. Eight DRB*W16 alleles and five DRB1*03 alleles were identified in this colony. We also identified two previously unreported DRB*W16 alleles, and confirmed inheritance of these alleles within several sibling groups. Subsequently, we investigated whether matching at MHC Class II DRB loci alone could predict alloreactivity, as assessed in vitro by two-way mixed lymphocyte reactions (MLRs). Fully DRB-matched, partially mismatched and fully mismatched animal pairs were prospectively chosen. MLR was performed using mononuclear cells (MNC) isolated from whole blood by density gradient separation. T-cell proliferation after 5-day culture was measured by (3)H-thymidine incorporation. Combined MNC from fully mismatched and partially mismatched animal pairs exhibited significant in vitro T-cell proliferation above single cell controls (P < 0.01). MNC from fully DRB-matched (but unrelated) animal pairs exhibited no proliferation compared with controls (P= 0.3). Using DRB genotyping, suitably alloreactive donor-recipient pairs may therefore be rapidly and accurately identified for use in further studies of cellular and solid organ transplantation.

Spontaneous glomerular mesangial lesions in common marmoset monkeys (Callithrix jacchus): a benign non‐progressive glomerulopathy

“…The development of a marmoset model of solid organ (particularly kidney) transplantation may be utilized to test clinically relevant therapies. As a prerequisite for this work, our group and others have developed a broad range of techniques including the rapid genotyping of marmoset major histocompatibility complex class II genes , the propagation of dendritic cells from precursors in bone marrow or peripheral blood , the establishment of the cross‐reactivity of human or NHP‐specific reagents with marmoset samples , and normal ranges for key hematological parameters.…”

“…The development of a marmoset model of solid organ (particularly kidney) transplantation may be utilized to test clinically relevant therapies. As a prerequisite for this work, our group and others have developed a broad range of techniques including the rapid genotyping of marmoset major histocompatibility complex class II genes [32,33], the propagation of dendritic cells from Spontaneous renal lesions have been reported in a range of Old World [15,18,39] and New World primates [3,17,31,39,41], including the common marmoset monkey [4-10, 19, 35, 37]. The nature of these has not been fully characterized, but in some cases, they have been attributed as a significant cause or contributor to animal death.…”

Comparative immunity of antigen recognition, differentiation, and other functional molecules: similarities and differences among common marmosets, humans, and mice

“…Plasad et al . reported the divergence of Caja-DRB and, specifically, the identification of DRB*W16 [ 64 , 65 ]. Similar to Caja-B, as reported by Shiina et al [ 77 ], classical HLA class II orthologs in CMs also appear to function similarly to human HLAs.…”

“…With regards to CM immunity, there are several previously reported research models. For example, there are models of infectious diseases [ 15 , 62 ], autoimmune disorders [ 1 , 6 , 30 , 84 ], and transplantation immunology [ 64 , 86 ]. CMs are susceptible to natural infections with particular strains of bacteria, which are thought to be caused by their diminished MHC class II repertoire [ 3 , 18 , 83 ].…”

Physiological and behavioral responses to routine procedures in captive common marmosets (Callithrix jacchus)

“…Callithrix jacchus, a small Neotropical primate endemic to the Brazilian Atlantic Rainforest (Stevenson and Rylands 1988), has been used for decades as a model species in a number of biomedical fields (Mansfield 2003;Prasad et al 2007; Kohu et al 2008). The species is commonly used as a test animal due to its good adaptation to captivity, high fertility rate, and relatively small size (Dixson and Lunn 1987;Lacreuse et al 2014).…”