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.
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.
Ouyang
Dongyun
D
Institute of Tissue Transplantation and Immunology, College of Life Science and Technology, Jinan University, Guangzhou 510632, China.
Wang
Xiaoying
X
He
Xianhui
X
Xu
Lihui
L
Shi
Huanjing
H
Gao
Qi
Q
Guo
He
H
D000818:Animals; D000956:Antigens, Viral / Q000276:immunology / Q000378:metabolism*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002681:China; D005822:Genetic Vectors; D015395:Histocompatibility Antigens Class I / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D016056:Immunodominant Epitopes / Q000276:immunology; D007194:India; D008253:Macaca mulatta; D010455:Peptides / Q000276:immunology / Q000378:metabolism*; D011485:Protein Binding; D015202:Protein Engineering; D017510:Protein Folding; D055503:Protein Multimerization; D015302:Simian Immunodeficiency Virus / Q000276:immunology*
D000956:Antigens, Viral; D015395:Histocompatibility Antigens Class I; D016056:Immunodominant Epitopes; C092249:Mamu-B 01 antigen, Macaca mulatta; D010455:Peptides