title: [Expansion and function of MHC restricted killer T cells derived from umbilical cord blood].
abstract: OBJECTIVE
  This study was to expand the cytotoxic T lymphocytes (CTL) through inducing the differentiation of umbilical blood monomuclear cells (UBMNC) by using various combination of cytokines, and to investigate the functions of expanded CTL.
  
  
  METHODS
  The MNC were isolated by ficoll density gradient centrifugation. Then, the PHA-P, IFN-γ combined with IL-2, IL-15 and other cytokines were used for induction and expansion of the cord blood-derived CTL. The biological function of CTL was examined by phenotype analysis, cytotoxic tests and real-time fluorescence quantitative PCR.
  
  
  RESULTS
  After expansion for 15 days, the cell number increased by 1522% ± 137%. The content of CD3(-)CD8(-) cells in uncultured cord blood MNC was 95%, and the CD3(+)CD8(+) CTL cells reached 82.77% in cultured cord blood MNC after expansion for 15 days. The expanded CTL cell showed the cytotoxic activity against K562 and HeLa cell line. The killing rate of MNC was 61.88 ± 1.08%. After expansion, the killing rate could reach to 90% with the average value of 90.33 ± 2.02%. The expanded CTL cells highly expressed some key cytokines, such as granzyme A, granzyme B, GM-CSF, granulysin, IFN-γ, TGF-β, TNF-α and perforin. Compared with the control group, the expression of IFN-γ and TGF-β significantly increased (P < 0.05), and the other factors dramatically increased (P < 0.01).
  
  
  CONCLUSIONS
  The cord blood-derived CTL can be expanded by different combinations of cytokines. These protocols may provide alternative choices for CTL cell expansion in tumor adoptive immunotherapy.
authors: Cord Blood Stem Cell Bank of Shandong Province, Jinan 250005, Shandong Province, China.
authors: Yi
authors: Y
authors: Ming
authors: Cord Blood Stem Cell Bank of Shandong Province, Jinan 250005, Shandong Province, China.
authors: Hai-Yan
authors: HY
authors: Zhang
authors: Department of Gynecology, Jinan Women and Children Health Care Hosital, Jinan 250001, Shandong Province, China.
authors: Zhe
authors: Z
authors: Wang
authors: Cord Blood Stem Cell Bank of Shandong Province, Jinan 250005, Shandong Province, China.
authors: Xiao-Dun
authors: XD
authors: Liu
authors: Cord Blood Stem Cell Bank of Shandong Province, Jinan 250005, Shandong Province, China.
authors: Chuan-Bao
authors: CB
authors: Zang
authors: Cord Blood Stem Cell Bank of Shandong Province, Jinan 250005, Shandong Province, China.
authors: Hang
authors: H
authors: Song
authors: Cord Blood Stem Cell Bank of Shandong Province, Jinan 250005, Shandong Province, China.
authors: Xiao-Yu
authors: XY
authors: Dai
authors: Cord Blood Stem Cell Bank of Shandong Province, Jinan 250005, Shandong Province, China; Cryomedicine Laboratory, Shandong University Qilu Hospital, Jinan 250001, Shandong Province, China. E-mail: lidong73@sdu.edu.cn.
authors: Dong
authors: D
authors: Li
mesh_terms: D016207:Cytokines; D005312:Fetal Blood; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D053804:Granzymes; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D006801:Humans; D016219:Immunotherapy, Adoptive; D054353:Perforin; D010835:Phytohemagglutinins; D013602:T-Lymphocytes, Cytotoxic
chemical_list: D016207:Cytokines; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D010835:Phytohemagglutinins; C028784:phytohemagglutinin-P; D054353:Perforin; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D053804:Granzymes