title: [Construction and primary application of two HBV-HLA-A*2402-peptide tetramers].
abstract: OBJECTIVE
  To construct two soluble HLA-A*2402-peptide tetramers and detect the HBV-specific cytotoxic T lymphocytes (CTLs) with the constructed HLA-A*2402-peptide tetramers.
  
  
  METHODS
  After proteins HLA-A*2402-BSP and beta2m were made an effective prokanyotical expression, the purified proteins were refolded into HLA-A*2402-beta2m-peptide complexes in the presence of two antigenic peptides (Hepatitis B virus Pol756-764 or Core117-125) with dilution method. Then the complexes were biotinylated by BirA enzyme and purified by gel-filtration chromatography. The tetramers were generated by mixing the complex with PE-Streptavidin in the molar ratio of 5:1. FCM can and cell quest software were used to analyze the stained PBMCs.
  
  
  RESULTS
  Two biotinylated HBV-HLA-A*2402-peptide complexes were identified by Western blot and they were shown to have natural conformation by Dot-ELISA and ELISA.
  
  
  CONCLUSIONS
  The constructed HLA-A*2402-peptide tetramers can detect the HBV-specific CTLs in the patients with self-limited acute HBV infection.
authors.affiliation: Division of Clinical Immunology, Tongji Hospital, Tongji Medical College, Huazhong Umiversity of Science and Technology, Wuhan, China. glp5177@126.com
authors.forename: Xin-Xing
authors.initials: XX
authors.lastname: Yang
authors.forename: You-Hua
authors.initials: YH
authors.lastname: Hao
authors.forename: Na
authors.initials: N
authors.lastname: Song
authors.forename: Ling
authors.initials: L
authors.lastname: Chen
authors.forename: Zhi
authors.initials: Z
authors.lastname: Liu
authors.forename: Dong-Liang
authors.initials: DL
authors.lastname: Yang
mesh_terms: D000818:Animals; D019921:Biotinylation; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D015234:HLA-A Antigens / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D059871:HLA-A24 Antigen; D006515:Hepatitis B virus / Q000276:immunology*; D006801:Humans; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D015234:HLA-A Antigens; C438506:HLA-A*24:02 antigen; D059871:HLA-A24 Antigen; D010455:Peptides