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Journal of Peking University (Health Sciences) ›› 2021, Vol. 53 ›› Issue (6): 1094-1098. doi: 10.19723/j.issn.1671-167X.2021.06.015

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Assessment of lymphocytic function in vitro stimulated by specific tumor polypeptide combined with dendritic cells

YANG Duo1,ZHOU Xin-na1,WANG Shuo1,WANG Xiao-li1,YUAN Yan-hua1,YANG Hua-bin1,GENG Hui-zhen2,PENG Bing2,LI Zi-bo2,LI Bin2,REN Jun1,△()   

  1. 1. Department of Oncology, Beijing Shijitan Hospital, Capital Medical University Beijing Key Laboratory of Therapeutic Vaccines, Beijing 100038, China
    2. Hebei Bio-High Technology Company Limited, Shijiazhuang 050035, China
  • Received:2021-02-04 Online:2021-12-18 Published:2021-12-13
  • Contact: Jun REN E-mail:jun.ren@duke.edu;renjun@bjsjth.edu.cn
  • Supported by:
    Special Project for the Construction of International Science and Technology Cooperation Base of Hebei Innovation Capability Improvement Plan Project(19392411D)

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Abstract:

Objective: To assess the activation function of specific tumor polypeptide for dendritic cell vaccine on lymphocytes proliferation, production of cytokines and killing activity in vitro by using dendritic cells as antigen presenting vector. Methods: Peripheral blood dendritic cells (DC) and cytokine-induced killer (CIK) were isolated and cultured by adherent culture method; CCK-8 method was used to assess the proliferation function of lymphocytes and the killing function of lymphocytes to tumor cells; enzyme-linked immunospot assay method was used to evaluate the secretion function of cytokines. The experiment was divided into tumor polypeptide group (peptide with DC-CIK), DC-CIK group and CIK group. Results: With presence of interleukin-2 (IL-2) in the culture system, the lymphocyte proliferation of the three groups was obvious. The absorbance at 450 nm of tumor polypeptide group was significantly higher than that of CIK group at the time points day 4 and day 6 (day 4: Z=-3.79, P<0.001; day 6: Z =-2.95, P< 0.01). The absorbance at 450 nm of group tumor polypeptide was significantly higher than that of DC-CIK group on day 4 (Z=-2.02, P<0.05). Without IL-2 in the culture system, lymphocytes proliferated slowly in all the three groups, and there was no significant difference in 450 nm absorbance at each time point. The levels of IL-4 (Z=-2.61, P< 0.01), granulocyte-macrophage colony-stimulation factor (GM-CSF, Z=-3.85, P< 0.001), interferon- γ (IFN- γ, Z=-3.56, P< 0.001) and tumor necrosis factor-α (TNF-ɑ, Z=-3.40, P< 0.001) of tumor polypeptide group were higher than those of CIK group. There was no significant difference in the production of cytokines except IL-4 (Z=-2.15, P< 0.05) when tumor polypeptide group was compared with DC-CIK group. The production of IFN-γ (Z=-2.44, P<0.05), TNF-ɑ (Z=-2.26, P< 0.05) and GM-CSF (Z=-3.73, P< 0.001) in DC-CIK group were higher than those of CIK group. Although there was no significant difference in killing activity between tumor polypeptide group, DC-CIK group and CIK group at hour 18 and hour 24, and the killing activity of tumor polypeptide group was higher than that of the other two groups. Conclusion: Tumor peptide combined with dendritic cells can improve the proliferation activity of CIK cells in vitro, and increase the secretion of several cytokines.

Key words: Dendritic cells, Tumor polypeptides, CIK cells, Proliferation, Cytokines, Cytotoxicity

CLC Number: 

  • R73-36+2

Table 1

Patient characters and specific tumor polypeptide information"

Serial number Gender Age/years Clinical diagnosis and stage Number of tumor specific
polypeptides*		 Polypeptide target
organ#		 Target cell△
1 Female 58 NSCLC stage Ⅳ 8 Lung A549
2 Male 59 Gastric cancer stage Ⅳ 4 Stomach SGC-7901
BGC-823
3 Female 57 Gastric cancer
stage Ⅳ		 6 Stomach SGC-7901
BGC-823
4 Male 64 Bladder cancer
stage Ⅳ		 3 Bladder TCCSUP
5 Female 73 Mammary cancer stage Ⅳ 5 Breast MCF-7
6 Female 56 NSCLC stage Ⅳ 6 Lung A549
7 Male 60 NSCLC stage Ⅳ 6 Lung A549
8 Female 46 NSCLC stage Ⅳ 8 Lung A549
9 Female 73 Pancreatic cancer stage Ⅳ 5 Pancreas CFPAC-1
10 Female 53 TNBC stage Ⅳ 6 Breast MDA-MB-453
11 Male 46 Colonic metastasis of gastric cancer stage Ⅳ 6 Colon HCT-116

Figure 1

Lymphocyte proliferation when stimulated by specific tumor polypeptides * P<0.05, # P<0.001. A, in the presence of IL-2, the absorbance at 450 nm of different groups at different time point; B, the absorbance at 450 nm of different groups at different time point without IL-2."

Figure 2

Cytokine secretion of CIK cells in different groups *P<0.05, △P<0.01, #P<0.001."

Figure 3

The killing efficiency of CIK cells to tumor cells at 18 h (A) and 24 h (B) time point with different effector/ target cells ratio"

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