Document Detail


Preparation and characterization of HLA-A*0201 tetramer loaded with IE-1316-324 antigenic peptide of human cytomegalovirus.
MedLine Citation:
PMID:  17092434     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Major histocompatibility complex (MHC) class I tetramer technology has become the central technique for analyzing antigen-specific CD8(+) T cell responses and it has been widely used to explore the differentiation and formation of memory CD8(+) T cells. Previously, a simplified and efficient procedure for preparing high quality HLA-A*0201 tetramers has been established in our lab and the tetramers loaded with HCMV peptide pp65(495-503) has been successfully applied to investigate HCMV-specific CD8(+) T cells in Chinese populations. Using similar procedure we reported here the construction of HLA-A*0201 tetramer loaded with another dominant epitope derived from immediate early (IE)-1(316-324) (VLEETSVML, VLE) of HCMV (A2-VLE) and characterization of this tetramer. After A2-VLE monomer was prepared and purified, its tetramer was then formed at a yield of 83%. The optimized amount of A2-VLE tetramer for staining 100 microl whole blood was 0.5 microg with incubation at 4 degrees C for 1 h. Furthermore, the dissociation constant of the tetramer binding to the specific CD8(+) T cells of one HLA-A2(+) donor was estimated to be 32.7 nmol/L, which is markedly higher than that of MHC monomer. The construction of A2-VLE tetramer provides an alternative choice for investigating HCMV-specific CD8(+) T cell responses and will deepen our understanding of the differentiation and formation of HCMV-specific memory CD8(+) T cells.
Authors:
Lihui Xu; Qingbing Zha; Hong Sun; Qiantao Jia; Fengyao Li; Xianhui He
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cellular & molecular immunology     Volume:  3     ISSN:  1672-7681     ISO Abbreviation:  Cell. Mol. Immunol.     Publication Date:  2006 Oct 
Date Detail:
Created Date:  2006-11-09     Completed Date:  2007-01-11     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101242872     Medline TA:  Cell Mol Immunol     Country:  China    
Other Details:
Languages:  eng     Pagination:  367-71     Citation Subset:  IM    
Affiliation:
Institute of Tissue Transplantation and Immunology, College of Life Science and Technology, Jinan University, Guangzhou 510632, China.
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MeSH Terms
Descriptor/Qualifier:
Biotin / chemistry
CD8-Positive T-Lymphocytes / immunology*
Cytomegalovirus / immunology*
Epitopes, T-Lymphocyte / immunology
Genes, MHC Class I / immunology
HLA-A Antigens / chemistry,  genetics,  immunology*
Humans
Immediate-Early Proteins / chemistry,  genetics,  immunology*
Phosphoproteins / chemistry,  genetics,  immunology
Protein Folding
Recombinant Proteins / chemistry,  genetics,  immunology
Viral Matrix Proteins / chemistry,  genetics,  immunology
Viral Proteins / chemistry,  genetics,  immunology*
Chemical
Reg. No./Substance:
0/Epitopes, T-Lymphocyte; 0/HLA-A Antigens; 0/HLA-A0201 antigen; 0/IE1 protein, cytomegalovirus; 0/IE1-pp65 recombinant protein, Human cytomegalovirus; 0/Immediate-Early Proteins; 0/Phosphoproteins; 0/Recombinant Proteins; 0/Viral Matrix Proteins; 0/Viral Proteins; 58-85-5/Biotin

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