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Artificial antigen presenting cells in vitro activate CD8+ T cells of hepatocellular carcinoma
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Abstract:
Objective:To explore the activation of CD8+T cells of hepatocellular carcinoma (HCC) stimulated by the artificial antigen presenting cells (aAPC) (K32/4-IBBL/CD86). Methods: Peripheral CD8+T cells of HLA-A*0201 positive HCC patients were enriched by MACS separation system. CD8+T cells were cultured with aAPC at different ratios (3∶1, 2∶1 and 1∶1), and CTLs were induced. Growth curve of CTLs was examined by trypan blue exclusion assay; MTS/PMS assay was used to detect the proliferation of CTLs; the IFN-γ secreting CTLs were assayed by FACS; the cytotoxicity of CTLs on a human liver cancer cell line BEL7402 and self-HCC cells was tested using MTT method. Results: The proliferation of peripheral CD8+ T cells stimulated with aAPC was significantly increased. At days 8, the CTL cell numbers of aAPC to CD8+ T cells at 1∶1, 1∶2, 1∶3 groups was significantly higher than those in the control group (\[212±2.5\]×105, \[17.6±3.2\]×105, \[15.3±2.8\]×105 vs \[8.5±0.15\]×105, P<0.01); and the proportion of IFN-γ secreting CTLs in 1∶1, 1∶2, 1∶3 groups was higher than those in the control group (\[26.2±1.91\]%, \[21.3±1.38\]%, \[18.6±1.20\]% vs \[0.1±0.02\]%, P<0.01). The cytotoxicity of aAPC-activated CTLs against BEL7402 cells and self-HCC cells in 1∶3 group was stronger than that in the control group (\[21.8±4.3\]% vs \[3.8±18\]%, P<0.01; \[25.6±3.6\]% vs \[3.8±2.0\]%, P<0.01). A higher cytotoxic rate on BEL7402 cells and self-HCC cells in the 1∶1 group at a ratio of E (effect): T (target) as 50∶1 was achieved compared with that in the 1∶2 and 1∶3 groups (\[568±3.0\]% vs \[44.3±2.6\]%, \[38.9±4.7\]%, P<0.05; \[64.8±4.2\]% vs \[56.1±3.4\]%, \[46.2±47\]%, P<0.05). Conclusion: aAPC can effectively activate CD8+T cells of HCC patients, and the resultant CTLs not only secrete IFN-γ but also efficiently kill human liver cancer BEL7402 cells and self-HCC cells.
Keywords:
Project Supported:
Project supported by the Scientific Research Foundation of Preventive Medicine of Health Bureau of Jiangsu Province（No.Y201032）
Clc Number:
