title: [Detection of IFN-gamma level in single CD8+ T cell].
abstract: OBJECTIVE
  To establish a new method capillary electrophoretic immunoassay laser-induced fluorescence (CEIA-LIF), to detect IFN-gamma level in single CD8(+) T cell.
  
  
  METHODS
  CD8(+) T cells were isolated from peripheral blood of two patients with severe aplastic anemia(SAA) and one normal person by Ficoll-Hypaque gradient centrifugation and then were purified by immunomagnetic microbead separation. Then purified CD8(+) T cells were incubated with digitonin for 15 min followed by FITC-anti-IFN-gamma mAb for 20 min. The single cell was detected continuously by CEIA-LIF. The feasibility of the method was confirmed by inverted microscopy and laser scanning confocal fluorescence microscopy.
  
  
  RESULTS
  The IFN-gamma content in purified CD8(+) T cells was detected under the condition of cell-membrane integrity. The IFN-gamma level in single CD8(+) T cell from 2 SAA patients was (151.53+/-28.92)zmol and (223.72+/-45.23)zmol, respectively, and much higher that from normal control (47.47+/-17.97)zmol ( P=0.001).
  
  
  CONCLUSIONS
  It is feasible to quantitate IFN-gamma in single CD8(+) T cell by CEIA-LIF. CEIA-LIF might be useful in the clinical detection of intracellular cytokines.
authors.affiliation: Department of Hematology, Qilu Hospital,Shandong University, Jinan 250012, China. jie_969@163.com
authors.forename: Zhi-Jie
authors.initials: ZJ
authors.lastname: Wang
authors.forename: Hua
authors.initials: H
authors.lastname: Zhang
authors.forename: Xue-Liang
authors.initials: XL
authors.lastname: Chen
authors.forename: Wen-Rui
authors.initials: WR
authors.lastname: Jin
authors.forename: Ya-Qin
authors.initials: YQ
authors.lastname: Cong
authors.forename: Bo
authors.initials: B
authors.lastname: Han
authors.forename: Yan
authors.initials: Y
authors.lastname: Gu
mesh_terms: D000936:Antigen-Antibody Complex / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D019075:Electrophoresis, Capillary; D006801:Humans; D007118:Immunoassay; D007371:Interferon-gamma / Q000032:analysis* / Q000276:immunology; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D013997:Time Factors
chemical_list: D000936:Antigen-Antibody Complex; D007371:Interferon-gamma