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Archive > Volume 16 Issue 1 > 2009,16(1):18-23. DOI:10.3872/j.issn.1007-385X.2009.1.005 Prev Next

Basic Research

CTL induced by NY-ESO-1 impulsed dendritic cells specifically kills NY-ESO-1 positive human hepatocellular caner cells

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    Abstract:
    Objective: To study the antigen specific antitumor effect of cytotoxic T lymphocyte (CTL), which was induced by cancer testis antigen NYESO1impulsed dendritic cells (DCs), against human hepatocellular carcinoma (HCC). Methods: GSTESO1 fusion protein was induced in recombinant pGEXESO1 vector transformed bacteria by IPTG, and the GSTESO1 fusion protein was purified. DCs were induced with granulocyte/macrophage colonystimulating factor (GMCSF) and interleukin4 (IL4) from human peripheral blood mononuclear cells. DCs impulsed with GSTESO protein peptide were cocultured with T lymphocytes, and the resultant CTLs were used as effector cells. NYESO1 positive hepatocellular carcinoma HepG2 cells and NYESO1 negative H2P cells were used as target cells to test the specific antitumor effect of CTL using MTT. Results: Escherichia coli BL21 expressed fusion protein peptide GSTESO1 (Mr 36 000) after transfection with recombinant pGEXESO1 vector. The concentration of GSTESO1 peptide was 50 μg/ml after purification. DCs were successfully induced with GMCSF and IL4 from human peripheral blood mononuclear cells. DCsimpulsed with NYESO1 had high expression of surface molecule such as HLADR(91.4%), CD86(70.5%), CD83(71.2%) and CD80(55.3%). DCsimpulsed with NYESO1 induced production and proliferation of CTL, and this CTL specifically killed NYESO1 positive HepG2 cells. CTL induced by NYESO1 had stronger cytotoxic effect against HepG2 cells compared with GSTimpulsed DCs, unimpulsed DCs (P<0.05). Highest antitumor activity was found when the ratio of effector∶target was 50∶1 (53.23±3.78, P<0.01). Conclusion: DCsimpulsed with NYESO1 can induce production and proliferation of allogenic CTLs, which show antigen specific antitumor effect against NYESO1 positive HCC cells. This result casts new lights on immunotherapy of HCC.

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    Project Supported:
    Supported by the Natural Science Foundation of Fujian Province（C0610023）；Science and Technology Planning Program of Guangzhou(No: 2003J1C0221)

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