Expansion in vitro and cytotoxicity of dendritic cells from patients with chronic myeloid leukemia.
- Author:
Lei JI
1
;
Pei-Ni XING
;
Xu-Cang WEI
;
Tong WANG
;
Mei-Sheng LI
;
Wang-Gang ZHANG
Author Information
1. Department of Hematology, The Second Hospital of Xi'an Jiaotong University, Xi'an 710004, China. jilei@163.com
- Publication Type:Journal Article
- MeSH:
Adolescent;
Adult;
Aged;
Antigens, CD34;
immunology;
Cell Proliferation;
Cells, Cultured;
Child;
Cytotoxicity, Immunologic;
Dendritic Cells;
immunology;
pathology;
Female;
Flow Cytometry;
Humans;
Immunophenotyping;
Leukemia, Myelogenous, Chronic, BCR-ABL Positive;
immunology;
pathology;
Male;
Middle Aged;
T-Lymphocytes, Cytotoxic;
immunology
- From:
Journal of Experimental Hematology
2005;13(2):198-204
- CountryChina
- Language:English
-
Abstract:
The study was aimed to investigate the extensive amplification and the cytotoxicity of dendritic cells (DC) derived from chronic myeloid leukemia cells. DC were cultured in two steps: firstly, extensive amplification in primary culture of CD34(+) or mononuclear cells isolated from CML patients' bone marrow and peripheral blood with rhFlt3-L and rhTPO for 7 days; secondly, inducing culture of DC with rhGM-CSF, rhTNF and rhIL-4 for 14 days. A system inducing DC directly were established for comparison. DC were identified by immunophenotype with flow cytometry, chromosome analysis by displaying G banding and electric microscopy analysis. The function of stimulating T cells proliferation and cytotoxicity of CML cells were confirmed through MTT assay. The results showed that after first extensive amplification in primary culture with rhFlt3-L and rhTPO for 7 days, CD34(+) cells had a total cell number with (77 +/- 5) fold expansion, and DC were (39 +/- 8)% of total cell respectively after induction culture of DC with rhGM-CSF, rhTNF and rhIL-4 for 14 days. Both the amplification of cell number and yield of DC were higher than the system without extensively culture (P < 0.01). Such DC could stimulate T cells to proliferate and kill leukemia cells finally. In conclusion, two-step culture method can obviously improve the cell number of DC required, that is better than inducing them directly. DC derived from CML cells induce the generation of anti-leukemia immunization.
