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Archive > Volume 20 Issue 6 > 2013,20(6):696-699. DOI:10.3872/j.issn.1007-385X.2013.06.011 Prev Next
Basic Research
Preparation of oligoclonal hepatocellular carcinoma-infiltrating lymphocytes and their cytotoxicity to autologous cancer cells
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Abstract:
Objective : To develop a novel method for isolation and culture of the oligoclonal tumor-infiltrating lymphocytes (TILs) with a stronger cytotoxic activity to autologous hepatoma carcinoma cells. Methods: Fresh human hepatocellular carcinoma (HCC) tissues were dissected to prepare conventional TILs and oligoclonal TILs by the way of enzymatic digestion with physical disaggregation and the new way of oligoclonal TILs culture from a single tumor fragment in microcultures. After 2-week culture, the oligoclonal TILs with a stronger cytotoxic activity were mergered for further expending. Different TILs-mediated lysis of HCC cells in vitro was determined by colorimetric tetrazolium (MTT) assay to compare the specific cytotoxicity between different oligoclonal TILs and between bulk TILs and oligoclonal TILs. Results: Lymphocytes could gradually migrate out of each tumor fragment and expand in the culture with high levels of IL-2. Oligoclonal TILs generated from different fragments showed different cytotoxic activities against autologous HCC cells, and the difference in cytotoxic activity between different oligoclonal TILs was significant (P<0.01). Moreover, the cytotoxic activity of oligoclonal TILs was significantly stronger than that of bulk TILs (\[72.56±6.69\]% vs \[46.24±403\]%, P<0.01). Conclusion: The cytotoxic activity against autologous HCC cells of oligoclonal TILs generated from HCC tissues is much stronger than that of conventional TILs.
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Project Supported:
Project supported by the National Natural Science Foundation of China （No. 81101716，No. 81272669），and the National Key Projects for Infectious Disease of China
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