Bioactivity of in vitro cultured tumor necrosis factor-alfa transduced tumor-infiltrating lymphocytes and therapeutic effects on human brain glioblastoma infused in different ways / 中国组织工程研究

ABSTRACT

BACKGROUND: Tumor-adopted immunity and gene transduction technique are used to introduce tumor necrosis factor-α vector into carrier cells, which are then re-infused into the body so that cancer cells can be killed by tumor necrosis factor-α more directly and effectively with fewer side effects on the other tissues due to high local expression.OBJECTIVE: To study the bioactivity of in vitro cultured tumor necrosis factor-α transduced tumor-infiltrating lymphocytes as well as the inhibitory effects on cancer cells of cancer-loaded rats infused in different ways.DESIGN: A randomized controlled study based on experimental animals.SEETING: Cancer Research Institute of China Medical University.MATERIALS: This study was carried out at the Cancer Research Institute and the Experimental Animal Department, China Medical University,between January 2000 and December 2001. TJ8510 cell line (human brain glioblastoma cell line) was provided by the Neurological Research Institute of Tianjin Medical University Affiliated Hospital. The experimental animals were 36 BALB/C nude mice congenitally having no thymius.METHODS: Based on the establishment of tumor necrosis factor-α retroviral transduction system and the preparation of cartier cells tumor-infil-trating lymphocytes, the monoclonal virus cell line PLC-2 and PLJC-5available were used to introduce marked gene NeoR and targeted gene tumor necrosis factor-α into tumor-infiltrating lymphocytes, respectively.Then cell proliferation, tumor necrosis factor expression and in vitro antitumor activity were examined. After cancer cell inoculation, the 36 nude mice were randomly divided into 6 groups: local infusion control group, local tumor-infiltrating lymphocytes infusion group, local tumor necrosis factor-tumor-infiltrating lymphocytes infusion group, venous infusion control group, venous tumor-infiltrating lymphocytes infusion group and venous tumor necrosis factor-tumor-infiltrating lymphocytes infusion group, and the therapeutic effects on the cancer-loaded mice were observed.proliferation and tumor necrosis factor-α expression in tumor-infiltrating oR-tumor-infiltrating lymphocytes and tumor necrosis factor-tumor-infiltrating lymphocytes was not significantly different from each other (P ＞ 0.05).NeoR-tumor-infiltrating lymphocytes, though not significantly different (P ＞0.05), significantly differ from that of tumor necrosis factor-tumor-infiltrating lymphocytes (P ＜ 0.01); moreover, tumor necrosis factor-tumor-infiltrating lymphocytes were found to express higher tumor necrosis factor-α conactivity did not significantly differ between tumor-infiltrating lymphocytes and NeoR-tumor-infiltrating lymphocytes (P ＞ 0.05), but obviously increased come of the animal experiment: 40 days after tumor necrosis factor-tumorinfiltrating lymphocytes infusion, cancer size in local tumor necrosis factortumor-infiltrating lymphocytes infusion group was found smaller than that in local infusion control group [(307±42) and (2 048±278) mm3, P ＜ 0.01],and it was also smaller in venous tumor necrosis factor-tumor-infiltrating lymphocytes infusion group than that in venous control group [(954±195)and (1 989±305) mm3 , P ＜ 0.05].CONCLUSION: Tumor necrosis factor-α gene transduced tumor-infiltrating lymphocytes could effectively express tumor necrosis factor, exerting higher and in vivo anti-tumor effects than tumor-infiltrating lymphocytes in cancer-loaded nude mice. No obvious inhibitory effects on the growth of subcutaneous solid carcinoma could be observed in nude mice after venous infusion of human brain glioblastoma tumor-infiltrating lymphocytes, but the inhibitory effects became obvious due to venous infusion of tumor necrosis factor-tumor-infiltrating lymphocytes and significant due to local tumor necrosis factor-tumor-infiltrating lymphocytes infusion, indicating that local infusion is the preferable way in the treatment of glioblastoma by immuno-gene therapy.