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Archive > Volume 20 Issue 3 > 2013,20(3):330-335. DOI:10.3872/j.issn.1007-385X.2013.03.013 Prev Next
Clinical Research
Effect of in vitro expansion on expressions of surface receptors and anti-tumor activity of NK cells derived from patients with esophageal cancer
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Abstract:
Objective:To explore the expressions of receptors before and after NK cell amplification from patients with esophageal cancer and its cytotoxicity to tumor cells. Methods: Peripheral blood was collected from the Fujian Provincial Tumor Hospital, including 20 cases of esophageal cancer patients and 10 cases of healthy donors (control group). NK cells were amplified by combination of IL-2+IL-12+IL-15+IL-18. Cell immunophenotype and NK cell receptor expressions (CD56+, CD69+，NKG2D, NKp30, NKp44, NKp46, CD158b and CD159a) were determined by flow cytometry. The cytotoxicity of NK cells to various tumor cell lines (K562, Raji, Eca-109 and TE-1) were detected by lactate dehydrogenase (LDH) assay. Results: Compared with the control group, the ratio of CD3+, CD4+and CD4+/CD8+T cells were significantly lower in the peripheral blood of patients with esophageal cancer (P<0.05), and the ratios of NK cells (CD56+) and regulatory T cells (Treg) were significantly higher (P<0.05).The ratio of NK cells (CD3-CD56+) increased up to 90% after being cultured in combination of IL-2+IL-12+IL-15+IL-18 for 20 days. NK cell count expanded up to 1 000 times (P<0.01). The ratios of CD3+, CD4+T cells, B cells (CD19), monocyte-macrophage cells (CD14) and regulatory T cells (T-reg) were in a significant reduction 20 days after culture (P<0.01), with no significant difference between the patients with esophageal cancer and the control group (P>0.05). CD69+and NK cell activating receptors (NKG2D, NKp30, NKp44, NKp46) were significantly increased and the inhibitory receptors (CD158b, CD159a) were significantly decreased (P<0.05). NK cells derived from patients with esophageal cancer showed significant increase of cytotoxicity on tumor cells K562, Raji, Eca-109 and TE-1 after culture for 20 days compared to those before culture (\[69.2±51\]% vs \[42.3±30\]%, \[44.6±3.2\]% vs \[21.1±2.0\]%, \[69.7±3.9\]% vs \[50.3±3.5\]%, \[67.1±4.5\]% vs \[41.2±3.3\]%, P<0.01). Conclusion:The combined cytokines of IL-2+IL-12+IL-15+IL-18 can effectively expand peripheral blood NK cells, up-regulate the expressions of activated receptor and down-regulate the expression of inhibitory receptors. The numbers and functions of the cultured NK cells can both meet the clinical treatment needs.
Keywords:
Project Supported:
Project supported by the Key Science Foundation of Science and Technology Bureau of Fujian Province (No. 2010Y0017, No.2008I0012)
Clc Number:
