| Preparation and characterization of HLA-A*0201 tetramer loaded with IE-1316-324 antigenic peptide of human cytomegalovirus. | |
MedLine Citation:
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PMID: 17092434 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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:
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Lihui Xu; Qingbing Zha; Hong Sun; Qiantao Jia; Fengyao Li; Xianhui He |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Cellular & molecular immunology Volume: 3 ISSN: 1672-7681 ISO Abbreviation: Cell. Mol. Immunol. Publication Date: 2006 Oct |
Date Detail:
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Created Date: 2006-11-09 Completed Date: 2007-01-11 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101242872 Medline TA: Cell Mol Immunol Country: China |
Other Details:
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Languages: eng Pagination: 367-71 Citation Subset: IM |
Affiliation:
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Institute of Tissue Transplantation and Immunology, College of Life Science and Technology, Jinan University, Guangzhou 510632, China. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Biotin
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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:
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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 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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