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Archive > Volume 20 Issue 6 > 2013,20(6):654-660. DOI:10.3872/j.issn.1007-385X.2013.06.004 Prev Next
Basic Research
Effect of combinatorial culture protocol (IL-2 and IL-5) on the proliferation of NK cells in the peripheral blood of breast cancer patients in vitro
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Abstract:
Objective:To observe the effect of culture with IL-2 and IL-15 in vitro on the proportion, the cell phenotype, the cytotoxic activity against tumor cells and the adhesion activity of NK cells, NKT cells and T cells subsets, and to discuss the possible mechanism. Methods:Five patients with breast cancer were obtained in the Department of Biotherapy, Tianjin Medical University Cancer Institute and Hospital from May 2012 to July 2012, and PBMCs of those patients were isolated by and cultured with the NK cells proliferation scheme contained IL-2+IL-5. The cell expansion fold and the proportion of different lymphocyte subsets were observed after cultured by IL-2 and IL-15 for 15 d. The changes of the cell immunophenotype and cell surface receptors were detected by flow cytometry. The anti-tumor cytotoxicity against HLA-match or HLA-mismatch cancer cells among three lymphocyte subsets were measured using LDH cytotoxicity assay and CD107a release assay. Adhesion between the total expansion products and HLA-match or HLA-mismatch tumor cells was detected by Live Cell Station. Results: After expansion by IL-2+IL-15 for 15 days, compared with preamplification, the proportions of NK cells (\[36.74±17.10\]% vs \[16.34±3.12\]%, P<0.05) and NKT cells (\[24.88±12.34\]% vs \[4.06±2.05\]%, P<0.05\] were significantly increased; the proportions of CD4+T cells and Treg cells were significantly decreased (P<0.05), the proportions of CD8+ T cells were significantly increased (P<0.05); the expressions of surface activation receptors NKp30 (\[92.38±7.19\]% vs (32.14±17.64)%, P<0.05\], NKp44 \[(74.26±1628\]% vs \[1.94±1.60\]%, P<0.05\] and NKG2D (\[98.58±1.28)% vs (66.50±24.84)%, P<0.05\] expressed on NK cells were increased significantly, while CD16 was obviously decreased (\[85.12±7.66\]% vs \[95.60±253\]%, P<0.05\]; the activation receptors DNAM-1 and NKG2D expressed on both NKT cells and T cells were significantly increased (P<0.05). The killing rates of total expansion cells, NK cells and NKT cells against HLA-mismatched tumor cells were significantly higher than those against HLA-matched tumor cells (P<0.05). In the 84th minute of co-incubation, the adherent number of total expansion cells combined to HLA-mismatched tumor cells were significantly higher than that of HLA-matched tumor cells (\[4.80±0.53\] vs \[2.25±0.35\], P<0.05\]. Conclusion: Not only NK cells but also NKT cells can be amplificated by cultured with IL-2 and IL-15, indicating that it can be used to expand CD56+ cells. Expansion products kill the tumor cells mainly by the NK cells killing activity without HLA-restricted.
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Project Supported:
Project supported by the National Key Basic Research Development Program（973 Program）of China （No.2012CB9333004） ，the National Natural Science Foundation of China（No.81072159）, and the Special Foundation for Clinical Trial of Cancer Hospital Affiliated to Tianjin Medical University（No.11L01）
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