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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
2
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pubmed:dateCreated |
2007-4-27
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pubmed:abstractText |
HLA-A* 2402 is one of the most frequently encountered HLA-A alleles in East Asian populations. In order to study the CD8+ T cell responses in Chinese populations, we have described the generation and functional test of HLA-A* 2402 tetramer loaded with HCMV pp65(341-349) peptide (QYDPVAALF, QYD). The cDNA of HLA-A* 2402 heavy chain was cloned by RT-PCR from one of the donors. DNA fragment encoding the ectodomain of HLA-A* 2402 heavy chain fused at its carboxyl-terminal a BirA substrate peptide (BSP) was amplified by PCR with the cloned heavy chain cDNA as a template. The wild-type gene of HLA-A* 2402-BSP was not expressed in Escherichia coli (E. coli), while mutant HLA-A* 2402-BSP gene with optimized codons was overexpressed as inclusion bodies in E. coli. Furthermore, the soluble HLA-A* 2402-QYD monomers were generated by in vitro refolding of washed inclusion bodies in the presence of beta2-microglobulin and QYD peptide. The tetramer was subsequently formed by mixing HLA-A* 2402-QYD monomers with streptavidin-PE at a molar ratio of 4:1. Flow cytometry analysis indicated that this tetramer possessed binding activity with specific CTL from HLA-A24+ donors and the frequencies of tetramer-binding CTL were 0.09% - 0.37% within total CD8+ T cells. This tetrameric agent provides a powerful tool to explore the secrets of CTL responses against HCMV antigens in HLA-A* 2402 individuals.
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pubmed:language |
chi
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Carbon-Nitrogen Ligases,
http://linkedlifedata.com/resource/pubmed/chemical/Escherichia coli Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/HLA-A*24:02 antigen,
http://linkedlifedata.com/resource/pubmed/chemical/HLA-A Antigens,
http://linkedlifedata.com/resource/pubmed/chemical/HLA-A24 Antigen,
http://linkedlifedata.com/resource/pubmed/chemical/Oligopeptides,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Fusion Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Repressor Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Viral Matrix Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/birA protein, E coli,
http://linkedlifedata.com/resource/pubmed/chemical/cytomegalovirus matrix protein 65kDa
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pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
1000-3061
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
23
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
284-91
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pubmed:dateRevised |
2011-11-17
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pubmed:meshHeading |
pubmed-meshheading:17460903-Amino Acid Sequence,
pubmed-meshheading:17460903-CD8-Positive T-Lymphocytes,
pubmed-meshheading:17460903-Carbon-Nitrogen Ligases,
pubmed-meshheading:17460903-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:17460903-Escherichia coli,
pubmed-meshheading:17460903-Escherichia coli Proteins,
pubmed-meshheading:17460903-Flow Cytometry,
pubmed-meshheading:17460903-Gene Expression,
pubmed-meshheading:17460903-HLA-A Antigens,
pubmed-meshheading:17460903-HLA-A24 Antigen,
pubmed-meshheading:17460903-Humans,
pubmed-meshheading:17460903-Oligopeptides,
pubmed-meshheading:17460903-Phosphoproteins,
pubmed-meshheading:17460903-Protein Multimerization,
pubmed-meshheading:17460903-Recombinant Fusion Proteins,
pubmed-meshheading:17460903-Repressor Proteins,
pubmed-meshheading:17460903-Substrate Specificity,
pubmed-meshheading:17460903-T-Lymphocytes, Cytotoxic,
pubmed-meshheading:17460903-Viral Matrix Proteins
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pubmed:year |
2007
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pubmed:articleTitle |
[Improved expression of HLA-A* 2402-BSP in Escherichia coli and its tetramer preparation].
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pubmed:affiliation |
Institute of Tissue Transplantation and Immunology, College of Life Science and Technology, Jinan University, Guangzhou 510632, China.
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pubmed:publicationType |
Journal Article,
English Abstract,
Research Support, Non-U.S. Gov't
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