title: [Amplification Ex Vivo and Cytocidal Activity of Leukemia Tumor-Associated Antigen-Specific Cytotoxic T Lymphocytes].
abstract: OBJECTIVE
  To explore the feasibility of amplifying the leukemia tumor associated antigens-specific cytotoxic T lymphocytes (TAA-CTL) ex vivo and to evaluate the cytotoxicity of TAA-CTL.
  
  
  METHODS
  The peripheral blood mononuclear cells were enriched by density gradient centrifugtion; TAA-CTL were generated by stimulation of PBMNC with peptide-pulsed DC at an effector-to-target ratio of 10:1; immunophenotype of TAA-CTL was analyzed by flow cytometry; cytotoxicity assay was used to evaluate the cytotoxic activity of TAA-CTL against peptide-pulsed autologous target cells (PHA-Blasts).
  
  
  RESULTS
  TAA-CTL expanded from volunteer showed a mean expansion of 3.81±1.61, the phenotyping of the TAA-CTL was predominantly CD3+ (97.22±0.71)% with varying content of CD4+ (41.47±27.08)% and CD8+ (56.40±11.15)% T cells, it also contained few nature killer cells (0.50±0.31)% and rare residual B cells (0.14±0.20)%; the subpopulations of TAA-CTL and CTL were not statisticaly significantly different in the proportion (P>0.05); the detection of intracellular cytokines after stimulation with peptide showed that the secretion rates of IFN-γ and TNF-α in CD8+ TAA-CTL were (27.67±2.21)% and (34.2±0.71)%, while the secretion rates were (21.6±2.55)% and (9.97±3.44)% in CD4+ TAA-CTL. Compared with the CD8+ TAA-CTL group, the secretion rates of IFN-γ and TNF-α were (1.36±0.04)% and (5.58±0.03)% in CD8+ CTL, the rates of IFN-γ and TNF-α were (0.91±0.06)% and (1.60±0.07)% in CD4+ CTL. The secretions of IFN-γ and TNF-α in CTL were both significantly lower than those in TAA-CTL (P<0.01); the specific killing efficiency of the TAA-CTL against TAA-pulesd target cells were (77.00±1.00)%, (67.40±3.60)%, (60.55±2.45)% and (26.85±5.25)%, when the effecto-target ratios were 40:1, 20:1, 10:1 and 5:1, and there was negligible lysis of TAA-CTL for PHA-blast (P<0.01).
  
  
  CONCLUSIONS
  TAA-CTL can be successfully induced and generated ex vivo from the healthy volunteer peripheral blood, and the TAA-CTL possess a specific killing activity.
authors.affiliation: Derparterment of Hematology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Derparterment of Hematology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China. E-mail: lxinahf@sina.com.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Derparterment of Hematology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China. E-mail: wangxingbing91@hotmail.com.
authors.forename: Xing-Bing
authors.initials: XB
authors.lastname: Wang
authors.affiliation: Derparterment of Hematology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.affiliation: Derparterment of Hematology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Li
authors.affiliation: Derparterment of Hematology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China.
authors.forename: Lu-Lu
authors.initials: LL
authors.lastname: Yang
authors.affiliation: Derparterment of Hematology, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Guo
mesh_terms: D000951:Antigens, Neoplasm; D001402:B-Lymphocytes; D016207:Cytokines; D006801:Humans; D007694:Killer Cells, Natural; D007938:Leukemia; D013602:T-Lymphocytes, Cytotoxic; D014409:Tumor Necrosis Factor-alpha
chemical_list: D000951:Antigens, Neoplasm; D016207:Cytokines; D014409:Tumor Necrosis Factor-alpha