PMID- 16686063
OWN - NLM
STAT- MEDLINE
DCOM- 20080220
LR  - 20060511
IS  - 0376-2491 (Print)
IS  - 0376-2491 (Linking)
VI  - 85
IP  - 49
DP  - 2005 Dec 28
TI  - [Immunotherapy for leukemia cells by using cytotoxic T lymphocyte specifically 
      against WT1-derived peptide: an experimental study in vitro].
PG  - 3475-80
AB  - OBJECTIVE: To investigate the effect of targeting immunotherapy for leukemia 
      cells by using cytotoxic T lymphocyte (CTL) specifically against WT (Wilm's 
      tumor) 1-derived peptide. METHODS: A 9-mer WT1 peptide (CMTWNQMNL) containing 
      HLA-A*0201-binding anchor motifs was synthesized. Dendritic cells (DCs) generated 
      from the peripheral blood mononuclear cells of an HLA-A*0201-positive healthy 
      donor were cultured and divided into 2 groups: experimental group to be loaded 
      with Wilms' tumor 1 (WT1) peptide so as to elicit CTL's specifically for WT1 
      peptide, restricted by HLA-A*0201, and control group. Six days later rhTNF-alpha 
      was added for 3 days more to promote the maturation of DCs. Before loading of WT1 
      peptide and 2 days after the addition of rhTNF-alpha direct immunofluorescence 
      labeling method was used with PE or FITC labeled mono-antibodies to detect the 
      expression of the surface antigens: CD83, CD1alpha, CD80, CD86, CD14, and HLA-DR. 
      DCs suspended and attached to wall were collected and then divided into 2 groups: 
      pure T cell group (group D) to be cultured in culture medium without IL-2, and 
      IL-2 + T cell group (Group C) to be cultured in 1640 culture medium with IL-2. 
      Eight days later the T cells of Group C were co-cultured with the DCs of the 
      experimental group (WTI peptide + DC + IL-2 + T cells, Group A) or the DCs of the 
      control group (DC + IL-2 + T cells, Group B). Five days later the killing 
      activity was detected. The CTLs of Groups A, B, and C at logarithmic growth phase 
      were mixed with leukemic cells of the lines: NB4/WT1D, NB4WT1A, NB4 (all 
      HLA-A*0201 +, WT1 +), U937 (HLA-A*0201 +, WTl -), and K562 (HLA-A*0201 -, WTI +), 
      and mononuclear cells of the bone marrow of leukemic patients at different 
      effector cell-target cell of 20:1 and 10:1. MTT method was used to examine the 
      killing rate of CTL to the target cells. RESULTS: (1) The killing rates of Group 
      A cells to NB4/WT1 D, NB4WTA, and NB4, leukemic cells were 60.4% +/- 3.1%, 56.4% 
      +/- 5.7%, and 55.0% +/- 3.7%, all significantly higher than those of the Group B 
      cells (10.9% +/- 1.6%, 11.1% +/- 2.7%, and 11.9% +/- 2.5%), and those of Group C 
      cells (9.1% +/- 1.0%, 9.2% +/- 1.7%, and 9.4% +/- 1.8%) (all P < 0.01). There 
      were no significant differences in the killing rates to U937 and K562 leukemic 
      cells among the 3 groups. (2) When the effect-target ratio was 20: 2, the killing 
      activity of the CTLs of Group A to the HLA-A*0201 +, WT1 + NB4/WT1 D, NB4/WT1A 
      and NB4 leukemic cells was significantly higher than those to the HLA-A*0201 +, 
      WT1 - U937 cells and the HLA-A*0201 -, WT1 + K562 cells (both P < 0.001), 
      however, not significantly different from that to the U937 and K562 cells. (3) 
      There were no significant differences in the killing activity of Group A cells to 
      NB4/WT1D, NB4/WT1A, and NB4 cells (P = 0. 065, P = 0.621). (4) When the 
      effect-target ratio was 10:2, the killing rates of Group A cells to the NB4/ 
      WT1D, NB4/WT1A, and NB4 cells were 45.9% +/- 3.9%, 43.9% +/- 3.7%, and 44.1% +/- 
      3.2% respectively, all significantly lower than those when the effect-target 
      ratio was 20:1 (all P < 0.01). CONCLUSION: CTLs specific for WT1 and restricted 
      by HLA-A*0201 exert specific lysis upon leukemia cell lines and primary leukemia 
      cells, but not upon normal hematopoietic cells, which provides a rationale for 
      developing a strategy of WT1 peptide-based adoptive T-cell therapy and 
      vaccination for human leukemia and solid tumors.
FAU - Gu, Wei-ying
AU  - Gu WY
AD  - Department of Hematology, Third Hospital Affiliated to Soochow University, 
      Changzhou 213003, China.
FAU - Cao, Xiang-shan
AU  - Cao XS
FAU - Qiu, Guo-qiang
AU  - Qiu GQ
FAU - Chen, Zi-xing
AU  - Chen ZX
FAU - Sheng, Li-xia
AU  - Sheng LX
FAU - Xie, Xiao-bao
AU  - Xie XB
FAU - He, Jun
AU  - He J
FAU - Cen, Jian-nong
AU  - Cen JN
FAU - Shen, Hui-ling
AU  - Shen HL
LA  - chi
PT  - English Abstract
PT  - Journal Article
PL  - China
TA  - Zhonghua Yi Xue Za Zhi
JT  - Zhonghua yi xue za zhi
JID - 7511141
RN  - 0 (HLA-A Antigens)
RN  - 0 (Oligopeptides)
RN  - 0 (WT1 Proteins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Binding Sites/genetics
MH  - Cell Line, Tumor
MH  - Dendritic Cells/cytology/immunology/metabolism
MH  - Flow Cytometry
MH  - HLA-A Antigens/genetics/metabolism
MH  - Humans
MH  - Immunophenotyping
MH  - Immunotherapy
MH  - K562 Cells
MH  - Leukemia/genetics/immunology/therapy
MH  - Oligopeptides/genetics/*immunology/metabolism
MH  - T-Lymphocytes, Cytotoxic/cytology/*immunology
MH  - Transfection
MH  - U937 Cells
MH  - WT1 Proteins/genetics/*immunology/metabolism
EDAT- 2006/05/12 09:00
MHDA- 2008/02/21 09:00
CRDT- 2006/05/12 09:00
PHST- 2006/05/12 09:00 [pubmed]
PHST- 2008/02/21 09:00 [medline]
PHST- 2006/05/12 09:00 [entrez]
PST - ppublish
SO  - Zhonghua Yi Xue Za Zhi. 2005 Dec 28;85(49):3475-80.