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Archive > Volume 23 Issue 2 > 2016,23(2):175-181. DOI:10.3872/j.issn.1007-385X.2016.02.004 Prev Next
Basic Research
Cytotoxic effect of CIK cells promoted by alpha-galactosylceramide pulsed dendritic cells against hepatoma carcinoma cells
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Abstract:
Objective:To investigate function and maturation of DCs pulsed by α-GalCer and to explore influence of co-culture of CIK cells with α-GalCer-DCs on phenotype, proliferation activity and efficiency of killing hepatic carcinoma of the CIK cells. Methods: DCs and CIK cells were induced from monocytes that separated from human peripheral blood by density gradient centrifugation. The suspension and adherent monocytes were cultured ex vivo to generate DC and CIK cells respectively. The phenotypes of DCs pulsed by α-GalCer were detected by flow cytometry (FCM), and mRNA expression of related genes in DCs were detected by qRT-PCR. As co-culture of CIK cells with α-GalCer-DCs, surface markers on the DC and the CIK cells were detected by FCM. Proliferation multiple of the CIK cells was examined with trypan blue staining; expression levels of genes related with function of the CIK cells were measured by Real-time PCR; and effect of α-GalCer-DCs on killing HepG2 cells by the CIK cells was tested with CCK-8 kit. Results: CIK cells and mature DCs could be obtained from induction of multiple cytokines; Excitation of α-GalCer could promote DCs maturation, increase the positive expressions of CD80, CD86, CD83 and CD11c (P<0.05, P<0.01) and mRNA levels of surface chemokine receptors CCR-7 , IL-12, and IL-10 in the DCs (P<0.05). Co-cuture of the CIK cells with α-GalCer could significantly increase expression of CD3+CD56+ and proliferation activity of the CIK cells (P<0.05, P<0.01), and mRNA expression levels of INF-γ, IL-12, perforin and particle enzyme B (P<0.05, P<0.01); Killing effects of the CIK, the DC-CIK and the α-GalCer-DC-CIK cells on HepG2 cells were enhanced with increase of effector-target ratio. At the same effector-target ratio, α-GalCer-DC-CIK cells had the highest cytotoxicity effect on HepG2 cells (P<0.05).Conclusion:Pulse of α-GalCer could promote the maturation of DCs. Co-culture of the α-GalCer-DC with CIK cells could enhance proliferation and maturation of CIK cells, and enhance its activity to kill hepatic carcinoma cells. Therefore, the works might provide experimental and theoretical basis for application of the DC-CIK in immunotherapy of carcinoma.
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Project Supported:
Project supported by the Science and Technology Development Plan of Shandong Province (No. 2012GGA01219), the Science and Technology Development Plan of Jinan City (No. 201302032), and the 2012 Innovation Fundation for Technology Based Firms of Jinan
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