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Archive > Volume 21 Issue 2 > 2014,21(2):147-152. DOI:10.3872/j.issn.1007-385X.2014.02.006 Prev Next
Basic Research
Effect of interleukin-27 on the proliferation and cytotoxicity of lymphadenoma cells of cytokine induced killer cells
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Abstract:
bjective : To investigate the effect of IL-27 on the proliferation and cytotoxicity of cytokine induced killer (CIK) cells derived from human peripheral blood mononuclear cells (PBMC). Methods: Monocytes were purified from PBMCs obtained from healthy adults. After being challenged with interferon-γ (IFN-γ) and anti-human CD3, the cells were radomly divided into six groups by different stimulating factors: A group (IL-2：1 000 U/ml), B group (IL-2：1 000 U/ml, IL-27：20 ng/ml), C group (IL-2：1 000 U/ml, IL-27：10 ng/ml), D group (IL-2：500 U/ml, IL-27：10 ng/ml), E group (IL-2：1 000 U/ml, IL-27：5 ng/ml), and F group (IL-2：1000 U/ml, IL-27：40 ng/ml). The morphology of the CIK cells in different groups were evaluated by inverted microscopy. The proportion of CIK cells respectively expressing CD3 +CD56 + and CD8 + were analyzed by fluorescence activating cell sorter (FACS) on days 7 , 9, 11 and 13 after treatments. The number of attached cells was counted by a computer-based cell counter. The cytotoxicity of CIK cells was determined by MTS test. Results: Compared with A, B and C groups, D group had significantly higher proportions of CD3 +CD56 + CIK cells (\[66.57±2.44\]% vs \[60.03±1.75\]%, \[5551±0.03\]%, and \[56.07±083\]%; P<0.05) and CD8 + CIK cells (\[81.67±1.97\]% vs \[70.30±2.67\]%, \[74.92±2.47\]%, and \[74.43±1.90\]%; P<0.05), and significantly higher index of cell expansion to culture median (4 811.87±23.07 vs 3 25773±91.97, 3 790.92±64.49, 4 009.85±43.08; P<0.05) on day 11 after treatments. The highest effector to target cell ratio on day 11 in D group was 40 ∶1 with a cytotoxicity rate of (76.71±221)% which was significantly higher than that in groups A, B, and C. Conclusion: In vitro, IL-27 is capable of significantly enhancing the proliferation and cytotoxicity of CIK cells in a time-dependent manner and the optimal time of incubation seems to be 11 days.
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Project Supported:
Projects supported by the National Natural Science Foundation of China(No.81201607), the Doctoral Foundation of New Teacher from Ministry of Education of China (No.20101323120004), and the National Natural Science Foundation of Hebei Province(No. H2012206135)
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