Characterization of major histocompatibility complex-related molecule 1 sequence variants in non-human primates
The major histocompatibility complex (MHC) class I-related molecule, MR1, presents vitamin B metabolites from bacteria and yeast to mucosal-associated invariant T (MAIT) cells. Despite the evolutionary conservation of MR1, we do not know whether different allele variants of MR1 exist within the nonhuman primate (NHP) populations that are commonly used for biomedical research. In this study, we identified 21 distinct MR1 nucleotide sequences representing 32 different alleles across five different NHP populations. The majority of the alleles conferring amino acid changes (allele variants) were found in or near the alpha-1 domain of the mature MR1 protein. We expressed four of the most commonly observed MR1 allele variants in 293T cells, and we found that each variant could present bacterial metabolites on the cell surface. We successfully induced cytokine production in macaque MAIT cells stimulated with 293T cells expressing the four most common MR1 allele variants, demonstrating the usefulness of these cell lines to study MAIT cell activity. Our data suggests that MR1 is not monomorphic, but that there are multiple MR1 alleles in NHPs. The materials we describe here will be valuable for characterizing differences in MR1 antigen presentation and MAIT cell function in NHPs.
KeywordsMHC class I-related molecule Nonhuman primate Population genetics Sequencing analysis
We thank Dane Gellerup and Katie Zarbock for technical support with sequencing experiments. We thank Julie Karl for help with the proper genetic nomenclature for the identified the MR1 alleles. We thank Sidonia Eckle and Erin Adams for their helpful comments and suggestions with regard to analyzing MR1 allele structure and function. We thank Dr. Stephen Kent (University of Melbourne; Melbourne, Australia) and Dr. JoAnne Flynn (University of Pittsburgh; Pittsburgh, Pennsylvania, USA) for providing frozen pigtailed macaque and Asian cynomolgus macaque PBMC samples, respectively. The MR1 tetramer technology was developed jointly by Dr. James McCluskey, Dr. Jamie Rossjohn, and Dr. David Fairlie, and the material was produced by the NIH Tetramer Core Facility as permitted to be distributed by the University of Melbourne.
The research was conducted in part at a facility constructed with support from Research Facilities Improvement Program grant numbers RR15459-01 and RR020141-01. The study was also supported by the members of the Wisconsin National Primate Research Center by grants P51RR000167 and P51OD011106. This study was funded in part by the National Institutes of Health NIH R21AI127127-01, and also in part by an NIH NIAID contract HHSN272201600007C.
Compliance with ethical standards
The marmosets used in this study were housed at the Wisconsin National Primate Research Center (WNPRC). They were cared for by staff at the Wisconsin National Primate Research Center using protocols approved by the University of Wisconsin Graduate School Animal Care and Use Committee.
Conflict of interest
The authors declare that they have no conflicts of interest.
All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.
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