Comparison of the proliferation, cytotoxic activity and cytokine secretion function of cascade primed immune cells and cytokine‑induced killer cells in vitro

Li,Gui‑Xin; Zhao,Shu‑Shu; Zhang,Xu‑Guang; Wang,Wen‑Hao; Liu,Jin; Xue,Ke‑Wei; Li,Xiao‑Yan; Guo,Ying‑Xue; Wang,Li‑Hua

doi:10.3892/mmr.2015.3765

August-2015 Volume 12 Issue 2

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August-2015 Volume 12 Issue 2

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Article Open Access

Comparison of the proliferation, cytotoxic activity and cytokine secretion function of cascade primed immune cells and cytokine‑induced killer cells in vitro

Authors:
- Gui‑Xin Li
- Shu‑Shu Zhao
- Xu‑Guang Zhang
- Wen‑Hao Wang
- Jin Liu
- Ke‑Wei Xue
- Xiao‑Yan Li
- Ying‑Xue Guo
- Li‑Hua Wang
View Affiliations / Copyright

Affiliations: Department of Oncology, Affiliated Hospital of Weifang Medical University, Weifang, Shandong 261031, P.R. China, Department of Clinical Laboratory, Affiliated Hospital of Weifang Medical University, Weifang, Shandong 261031, P.R. China

Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].
Pages: 2629-2635
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Published online on: May 8, 2015

https://doi.org/10.3892/mmr.2015.3765
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Abstract

The present study aimed to compare the antitumor effects of cascade primed immune (CAPRI) cells and cytokine‑induced killer (CIK) cells in vitro, through investigating cell morphology, proliferation, cytotoxic activity to tumor cells and the ability of these cells to secrete cytokines. Peripheral blood samples (50 ml) were obtained from three healthy volunteers and peripheral blood mononuclear cells (PBMCs) were obtained from each via Ficoll‑Conray density gradient centrifugation. Each suspension of PBMCs (1x106/ml) was divided into two parts; CAPRI cells were obtained from one part through a series of induction, amplification and cytokine cultures, while CIK cells were obtained from the other part through induction with different cytokines. During the culture process, the proliferation and morphological changes were observed for the two cell types using Trypan blue staining. At day 14, the cytotoxic activity of the two cell types was examined through determining lactate dehydrogenase release in the presence of K562 leukemia cells and MCF‑7 breast cancer cells. In addition, secretory levels of interferon (IFN)‑γ and interleukin (IL)‑2 were detected using enzyme-linked immunospot (ELISPOT) technology. The results revealed that at day 5 and 14 of culture, there were significantly fewer CAPRI cells compared with CIK cells (P<0.001), although the survival rate of each cell type was >95%. The cytotoxic activity of CAPRI cells towards the K562 cell line was effector‑target ratio‑dependent (40:1 and 20:1) with values of 55.1±3.25 and 35.0±2.65%, respectively, which were significantly reduced compared with the corresponding data in CIK cells, 60.0±3.03 and 39.7±3.42% (P=0.004 and 0.005, respectively). Furthermore, the cytotoxic activity of CAPRI cells towards MCF‑7 cells were 71.5±3.06, 56.0±3.76 and 40.2±2.90% at effector‑target ratios 40:1, 20:1 and 10:1, respectively. These data were significantly higher than the corresponding values in CIK cells, 65.4±3.86, 49.5±3.91 and 36.1±3.73% (P=0.002, 0.003 and 0.02, respectively). As determined using ELISPOT technology at different cell concentrations (1x106/ml and 5x105/ml), IFN‑γ secretion levels, determined by the number of spot‑forming cells, of CAPRI cells were 126.2±10.31 and 48.8±10.99, respectively, which were significantly reduced compared with those of CIK cells, 409.3±7.76 and 159.3±15.45, respectively (P<0.001). IL‑2 secretion levels in CAPRI cells were 325.1±16.24 and 113.8±11.29 at 1x106/ml and 5x105/ml, respectively, which were significantly increased compared with CIK cells, 212.0±16.58 and 70.7±10.57, respectively (P<0.001). In conclusion, the present study demonstrated that CAPRI cells had a reduced proliferation rate compared with CIK cells as well as a less potent cytotoxic effect on K562 cells; however, the two cell types had potent cytotoxic activity towards solid tumor MCF‑7 cells. In addition, CAPRI cells secreted lower levels of IFN‑γ and increased levels of IL‑2 compared with CIK cells. These results indicated that antitumor activities of CAPRI and CIK cells proceeded via different mechanisms.

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1	Dong Z, Qiao Y, Li L, et al: Report of Chinese cancer control strategy. Bull Chin Cancer. 11:250–260. 2002.
2	Tang ZY: Modern Oncology. 2nd. Shanghai Medical University Press; Shanghai: pp. 513–516. 2000
3	Han BH: Tumor Biological Immune Targeted Therapy. 1st. Shanghai Science and Technology Publishing House; Shanghai: pp. 47–52. 2006
4	Hao: Solid Tumor Cellular Immunotherapy. Beijing: People’s Medical Publishing House; Beijing: pp. 3–4. 2010
5	Sha W: Clinical Tumor Biological Immunotherapy. 1st. Tianjin Science and Technology Press; Tianjin: pp. 142–144. 2006
6	Laumbacher B, Gu S and Wank R: Activated monocytes prime naïve T cells against autologous cancer: vigorous cancer destruction in vitro and in vivo. Scand J Immunol. 75:314–328. 2012. View Article : Google Scholar
7	Ren H, Xing S, Li D, et al: The proliferation profile in vitro and anti-tumor effects of CIK cells in vivo and in vitro. Chin J Cancer Biother. 6:17–21. 1999.
8	Morse MA, Glay TM and Lyerly HK: Current status of adoptive immunotherapy of malignancies. Expert Opin Biol Ther. 2:237–247. 2002. View Article : Google Scholar : PubMed/NCBI
9	Tai G: Tumor immunotherapy: Progresses and trends. Chin J Cancer Biother. 16:427–430. 2009.
10	Anne C, Eaton Armstrong David, Ewing Joanne C, et al: Cellular immunotherapy for cancer. BMJ Chinese Edition. 3:331–335. 2003.
11	Yang JJ, Xu XX and Qian G: Research progress on tumor biochemotherapy. Int J Respir. 30:1137–1141. 2010.
12	Qian QJ and Wu MC: Adoptive cell therapy of cancer-An old story with a new twist. Chin J Cancer Biother. 18:1–6. 2011.
13	Pan CC, Huang ZL, Li W, Zhao M, Zhou QM, Xia JC and Wu PH: Serum alpha-fetoprotein measurement in predicting clinical outcome related to autologous cytokine-induced killer cells in patients with hepatocellular carcinoma undergone minimally invasive therapy. Chin J Cancer. 29:596–602. 2010. View Article : Google Scholar : PubMed/NCBI
14	Li H, Yu JP, Cao S, Wei F, Zhang P, An XM, Huang ZT and Ren XB: CD4+ CD25+ regulatory T cells decreased the antitumor activity of cytokine-induced killer (CIK) cells of lung cancer patients. J Clin Immunol. 27:317–326. 2007. View Article : Google Scholar : PubMed/NCBI
15	Mehta BA, Schmidt-Wolf IG, Weissman IL and Negrin RS: Two pathways of exocytosis of cytoplasmic granule contents and target cell killing by cytokine-induced CD3+ CD56+ killer cells. Blood. 86:3493–3499. 1995.PubMed/NCBI
16	Ren H, Xing SX, Xu GW, et al: In vitro proliferation of CIK and kill tumor in vivo and in vitro activity of experimental study. Chinese journal of tumor biological treatment. 6:17–21. 1999.
17	Linn YC, Wang SM and Hui KM: Comparative gene expression profiling of cytokine-induced killer cells in response to acute myloid leukemic and acute lymphoblastic leukemic stimulators using oligonucleotide arrays. Exp Hematol. 33:671–681. 2005. View Article : Google Scholar : PubMed/NCBI
18	Sui CG, Meng F, Wang XH, et al: Preparation and cytotokic effects of cytokine-induced killer cells on different tumor cell lines in vitro. J Chin Med Univ. 34:210–211. 2005.
19	Shirai A, Homes K and Klinman D: Detection and quantitation of cells secreting IL-6 under physiologic conditions in BALB/c mice. J Immunol. 150:793–799. 1993.PubMed/NCBI
20	Yang J, Lemas VM, Flinn IW, Krone C and Ambinder RF: Application of the ELISPOT assay to the characterization of CD8(+) responses to Epstein-Barr virus antigens. Blood. 95:241–248. 2000.
21	Garrido F and Ruiz-Cabello F: MHC expression on human tumors - its relevance for local tumor growth and metastasis. Semin Cancer Biol. 2:3–10. 1991.PubMed/NCBI
22	van der Merwe PA and Davis SJ: Molecular interactions mediating T cell antigen recognition. Annu Rev Immunol. 21:659–684. 2003. View Article : Google Scholar : PubMed/NCBI
23	Verneris MR, Kornacker M, Mailänder V and Negrin RS: Resistance of ex vivo expanded CD3+CD56+ T cells to Fas-mediated apoptosis. Cancer Immunol Immunothr. 49:335–345. 2000. View Article : Google Scholar
24	Schmidt-Wolf IG, Finke S, Trojaneck B, et al: Phase I clinical study applying autologous immunological effector cells transfected with the interleukin-2 gene in patients with metastatic renal cancer, colorectal cancer and lymphoma. Br J Cancer. 81:1009–1016. 1999. View Article : Google Scholar : PubMed/NCBI
25	Jiang HN, Qi YC and Wang YD: Character and advancement of cytokine-induced kill cells. Tum J World. 4:240–242. 2005.
26	Lei J, Qian M, Wu ZL, Zhang SQ and Yan YC: Study on cytokine expression profiles of CIK cells in vitro. Chin J Immunol. 21:589–593. 2005.
27	Laport GG, Sheehan K, Baker J, et al: Adoptive immunotherapy with cytokine-induced killer cells for patients with relapsed hematologic malignancies after allogeneic hematopoietic cell transplantation. Biol Blood Marrow Transplant. 17:1679–1687. 2011. View Article : Google Scholar : PubMed/NCBI