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应用了小鼠B16F10黑色素瘤系统,观察到经基因转导后分泌IL-2的肿瘤细胞的排斥反应,发现此排斥反应主要巾巨噬细胞而非T、NK细胞介导。用逆转录病毒载体DC/TKIL2转染小鼠黑色素瘤细胞系B16F10,挑选出分泌相对高水平IL-2的克隆F10/IL2 Hi及低水平IL-2的克隆F10/IL2 Lo,其细胞增殖能力未发生变化。分别注射同系C57BL/6小鼠皮下,10至15天后接种亲本及转导对照载体DCA的F10DC细胞的对照组小鼠大多山现肿瘤且迅速生长;接种F10/IL2 Lo细胞的小鼠肿瘤的生长较对照组小鼠明显缓慢;接种F10/IL2 Hi细胞的小鼠肿瘤出现延迟,其中1/4小鼠在40至50天后仍未出现肿瘤,此组小鼠中排斥肿瘤者再以亲本B16F10细胞攻击,亲本肿瘤细胞生长未受明显抑制,表明小鼠不具有抗亲本细胞的保护力。用逆转录病毒载体将IFN-γ基因再转入F10/IL2Hi细胞,此双重转导细胞能表达MHC Ⅰ类Ⅱ类抗原且IL-2分泌能力未改变,接种小鼠后,其肿瘤生长情况与F10/IL2 Hi肿瘤生长情况无差异,提示在体内肿瘤细胞表达这些MHC抗原不能进一步增强肿瘤排斥,也提示有T细胞之外的细胞参与肿瘤的排斥。进一步用单抗去除小鼠体内CD8~+、NK1.1+细胞后接种分泌IL-2的肿瘤细胞,发现其较未去除组小鼠肿瘤生长无显著差异;在SCID和nu/nu小鼠也见到F10/IL2Hi细胞的排斥;?
Using mouse B16F10 melanoma system, the rejection of IL-2 secreting tumor cells after gene transduction was observed. It was found that this rejection was mainly mediated by macrophage rather than T and NK cells. The mouse melanoma cell line B16F10 was transfected with the retroviral vector DC/TKIL2 and cloned F10/IL2 Lo, a clone that secretes a relatively high level of IL-2, and F10/IL2 Lo, a low level of IL-2, and cell proliferation ability. No change has occurred. The mice in the control group were inoculated subcutaneously with syngeneic C57BL/6 mice subcutaneously and inoculated with parental and F10DC cells transduced with the control vector DCA for 10 to 15 days. Most of the mice in the control group grew rapidly and grew; tumors of mice inoculated with F10/IL2 Lo cells were grown. The mice were significantly slower than those in the control group; tumors in mice inoculated with F10/IL2 Hi cells were delayed, and 1/4 of the mice did not develop tumors after 40 to 50 days. In this group of mice, tumor rejection was attributed to the parental B16F10 cells. Upon challenge, the growth of the parental tumor cells was not significantly inhibited, indicating that the mice do not have protective power against the parental cells. The IFN-γ gene was retransfected into F10/IL2Hi cells with a retrovirus vector. The double transduced cells can express MHC class I antigens and the IL-2 secretion ability does not change. After inoculation of mice, the tumor growth and There was no difference in the growth of F10/IL2 Hi tumors, suggesting that the expression of these MHC antigens in tumor cells in vivo does not further enhance tumor rejection, suggesting that cells other than T cells participate in tumor rejection. After further removing the CD8~+ and NK1.1+ cells in mice with a monoclonal antibody and inoculating IL-2 secreting tumor cells, it was found that there was no significant difference in tumor growth between the mice in the untreated group and the mice in SCID and nu/nu mice. See the rejection of F10/IL2Hi cells;