Construction of soluble Mamu-b*1703, a class I major histocompatibility complex of Chinese rhesus macaques, monomer and tetramer loaded with a simian immunodeficiency virus peptide

Cell Mol Immunol. 2009 Apr;6(2):117-22. doi: 10.1038/cmi.2009.16.


Chinese-descent rhesus macaques have become more prevalent for HIV infection and vaccine investigation than Indian-origin macaques. Most of the currently available data and reagents such as major histocompatibility complex (MHC) class I tetramers, however, were derived from Indian-origin macaques due to the dominant use of these animals in history. Although there are significant differences in the immunogenetic background between the two macaque populations, they share a few of common MHC class I alleles. We reported in this study the procedure for preparation of a soluble Mamu-B*1703 (a MHC class I molecule of Chinese macaques) monomer and tetramer loaded with a dominant simian immunodeficiency virus (SIV) epitope IW9 (IRYPKTFGW) that was identified to be Mamu-B*1701-restricted in Indian macaques. The DNA fragment encoding the Mamu-B*1703 extracellular domain fused with a BirA substrate peptide (BSP) was amplified from a previously cloned cDNA and inserted into a prokaratic expression vector. In the presence of the antigenic peptide IW9 and light chain beta2-microglobulin, the expressed heavy chain was refolded into a soluble monomer. After biotinylation, four monomers were polymerized as a tetramer by phycoerythrin-conjugated streptavidin. The tetramer, having been confirmed to have the right conformation, was a potential tool for investigation of antigen-specific CD8(+) T-lymphocytes in SIV vaccine models of Chinese macaques. And our results also suggested that some antigenic peptides reported in Indian-origin macaques could be directly recruited as ligands for construction of Chinese macaque MHC tetramers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Viral / immunology
  • Antigens, Viral / metabolism*
  • CD8-Positive T-Lymphocytes / immunology
  • China
  • Genetic Vectors
  • Histocompatibility Antigens Class I / genetics*
  • Histocompatibility Antigens Class I / immunology
  • Histocompatibility Antigens Class I / metabolism
  • Immunodominant Epitopes / immunology
  • India
  • Macaca mulatta
  • Peptides / immunology
  • Peptides / metabolism*
  • Protein Binding
  • Protein Engineering
  • Protein Folding
  • Protein Multimerization
  • Simian Immunodeficiency Virus / immunology*


  • Antigens, Viral
  • Histocompatibility Antigens Class I
  • Immunodominant Epitopes
  • Mamu-B 01 antigen, Macaca mulatta
  • Peptides