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Updated: Jul 16, 2026
Analysis of Simian Immunodeficiency Virus-specific CD8+ T-cells in Rhesus Macaques by Peptide-MHC-I Tetramer Staining
Published on: December 23, 2016
Reference strand-mediated conformation analysis-based typing of multiple alleles in the rhesus macaque MHC class I
Yumiko Tanaka-Takahashi1, Michio Yasunami, Taeko Naruse
1Department of Molecular Pathogenesis, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.
Abstract:
The rhesus macaque exhibits individual differences in susceptibility and resistance to infectious agents such as simian immunodeficiency virus (SIV) under experimental conditions, and these may be genetically determined at least in part by major histocompatibility complex (MHC) class I polymorphism. Although the importance of defining MHC class I polymorphism is well recognized, development of a generic and comprehensive molecular typing method of MHC class I alleles of the rhesus macaque has been hampered because, during the evolution of this species, multiple copies of similar DNA sequences have been generated by duplication events including the coding sequences of Mamu-A and Mamu-B loci. We report here a newly developed reference strand-mediated conformation analysis (RSCA)-based typing method of multiple Mamu-A and Mamu-B cDNAs that allowed us to estimate the number of expressed alleles. This technique detected 1-7 Mamu-A signals and 2-12 Mamu-B signals in a single sample, indicating that the number of functional alleles may vary. By comparing the data from the parents with those from the descendants in the breeding colony, several MHC class I haplotypes consisting of variable numbers of functional Mamu-A and Mamu-B alleles could be assigned.
Insights
Rhesus macaques show varied resistance to infections due to genetic factors, particularly Major Histocompatibility Complex (MHC) class I. A new typing method reveals diverse numbers of Mamu-A and Mamu-B alleles, aiding genetic studies.
Area of Science:
- Immunogenetics
- Primate genomics
- Molecular typing
Background:
- Individual differences in rhesus macaque susceptibility to pathogens like SIV are partly genetically determined by MHC class I.
- Defining MHC class I polymorphism is crucial but challenging due to gene duplication events in rhesus macaques.
- Existing methods struggle with the complexity of Mamu-A and Mamu-B loci.
Purpose of the Study:
- To develop a comprehensive molecular typing method for rhesus macaque MHC class I alleles (Mamu-A and Mamu-B).
- To accurately estimate the number of expressed Mamu-A and Mamu-B alleles in individual animals.
- To facilitate the assignment of MHC class I haplotypes in a rhesus macaque breeding colony.
Main Methods:
- Development of a novel reference strand-mediated conformation analysis (RSCA) technique.
- Application of RSCA for typing multiple Mamu-A and Mamu-B cDNAs.
- Comparative analysis of typing data from parents and offspring in a breeding colony.
Main Results:
- The RSCA method successfully typed multiple Mamu-A and Mamu-B cDNAs.
- Detected a variable number of expressed alleles per sample: 1-7 Mamu-A signals and 2-12 Mamu-B signals.
- Successfully assigned several MHC class I haplotypes, revealing variable allele numbers per haplotype.
Conclusions:
- The new RSCA-based method provides a generic and comprehensive approach for typing rhesus macaque MHC class I alleles.
- The findings highlight significant variation in the number of functional Mamu-A and Mamu-B alleles, impacting immunogenetic studies.
- This method advances our ability to study the genetic basis of disease resistance in rhesus macaques.
