IL-15基因轉(zhuǎn)染人CIK細(xì)胞過繼轉(zhuǎn)移治療胃癌的實(shí)驗(yàn)研究
發(fā)布時(shí)間：2018-11-24 11:30
【摘要】： 本實(shí)驗(yàn)構(gòu)建了hIL-15-IRES-TK逆轉(zhuǎn)錄病毒載體,并證實(shí)可有效轉(zhuǎn)染CIK細(xì)胞。其轉(zhuǎn)染可使CIK細(xì)胞自分泌白介素-15(IL-15)。鑒于逆轉(zhuǎn)錄病毒轉(zhuǎn)染具有潛在的導(dǎo)致自身免疫疾病的危險(xiǎn),同時(shí)表達(dá)的胸苷激酶基因(HSV-TK)可使被轉(zhuǎn)染的CIK細(xì)胞在丙氧烏苷(GCV)的處理下能被有效清除。這為hIL-15-TK病毒載體應(yīng)用于臨床CIK細(xì)胞過繼轉(zhuǎn)移治療腫瘤的安全性提供了重要的基礎(chǔ)。 為了觀察hIL15-IRES-TK質(zhì)粒轉(zhuǎn)染的人源T細(xì)胞在體內(nèi)治療人源胃腸道腫瘤的療效,我們利用T細(xì)胞免疫缺陷小鼠(Balb/c-nu/nu鼠)以人的低分化胃腺癌細(xì)胞株BGC-823成功制備了人源胃癌細(xì)胞荷瘤小鼠模型,分別以hIL-15-IRES-TK逆轉(zhuǎn)錄病毒轉(zhuǎn)染和未轉(zhuǎn)染的CIK細(xì)胞在瘤體局部注射,1次/周,共2周,治療后觀察2周。以生理鹽水局部注射作為空白對(duì)照組。每周測(cè)量裸鼠腫瘤體積1次,4周后處死裸鼠,取出腫瘤組織稱重。 經(jīng)過2周的治療,分別注射hIL-15-IRES-TK質(zhì)粒轉(zhuǎn)染與非轉(zhuǎn)染的CIK細(xì)胞的兩組荷瘤小鼠的腫瘤體積出現(xiàn)了顯著性差異,顯示出hIL-15-IRES-TK質(zhì)粒轉(zhuǎn)染的CIK細(xì)胞比非轉(zhuǎn)染的CIK細(xì)胞有更強(qiáng)的抑制腫瘤增長的活性。在第3、4周,停止CIK治療后3組小鼠的腫瘤體積均有所增加,但hIL-15-IRES-TK質(zhì)粒轉(zhuǎn)染與非轉(zhuǎn)染的CIK細(xì)胞組均小于生理鹽水組,說明CIK細(xì)胞過繼轉(zhuǎn)移治療具備一定的抑制腫瘤細(xì)胞的作用。而hIL-15-IRES-TK質(zhì)粒轉(zhuǎn)染與非轉(zhuǎn)染的CIK細(xì)胞兩組小鼠的腫瘤體積差異依然顯著,顯示出hIL-15-IRES-TK質(zhì)粒轉(zhuǎn)染的CIK細(xì)胞比非轉(zhuǎn)染的CIK細(xì)胞抑制腫瘤增長的時(shí)效更長。 在實(shí)驗(yàn)過程中IL-15轉(zhuǎn)染CIK組開始治療后有兩只裸鼠腫瘤潰爛,第二周開始潰爛結(jié)疤,第四周潰爛愈合,腫瘤消失。對(duì)照組與CIK組則無此現(xiàn)象出現(xiàn)。這一現(xiàn)象也側(cè)面證實(shí)了IL-15-IRES-TK質(zhì)粒轉(zhuǎn)染的CIK細(xì)胞比非轉(zhuǎn)染的CIK細(xì)胞具備更強(qiáng)和更持續(xù)的殺瘤效應(yīng)。 結(jié)論:實(shí)驗(yàn)表明CIK細(xì)胞可以有效抑制胃腺癌細(xì)胞株BGC-823在體內(nèi)的生長,而hIL-15-IRES-TK質(zhì)粒轉(zhuǎn)染的CIK細(xì)胞不僅可以抑瘤,還可以致腫瘤消退。
[Abstract]:In this study, hIL-15-IRES-TK retrovirus vector was constructed, and it was confirmed that it could be transfected into CIK cells effectively. The transfection can make CIK cells secrete interleukin-15 (IL-15). In view of the potential risk of autoimmune disease caused by retrovirus transfection, the expressed thymidine kinase gene (HSV-TK) can effectively remove the transfected CIK cells under the treatment of (GCV). This provides an important basis for the safety of hIL-15-TK virus vector in clinical CIK cell adoptive metastasis. To observe the therapeutic effect of human T cells transfected with hIL15-IRES-TK plasmid on human gastrointestinal carcinoma in vivo. We successfully established a tumor-bearing mouse model of human gastric cancer cells by using T cell immunodeficient mice (Balb/c-nu/nu mice) with human poorly differentiated gastric adenocarcinoma cell line BGC-823. HIL-15-IRES-TK retrovirus-transfected and untransfected CIK cells were injected locally into the tumor once a week for 2 weeks. Saline injection was used as the blank control group. The tumor volume of nude mice was measured once a week. After 4 weeks, the nude mice were killed and the tumor tissue was weighed. After two weeks of treatment, there was a significant difference in tumor volume between the two groups injected with hIL-15-IRES-TK plasmid and non-transfected CIK cells, respectively. The results showed that CIK cells transfected with hIL-15-IRES-TK plasmid had stronger inhibitory activity against tumor growth than non-transfected CIK cells. At the 3rd week, the tumor volume of the three groups increased after CIK treatment, but the hIL-15-IRES-TK plasmid transfection and non-transfection CIK cells were smaller than the normal saline group. These results suggest that adoptive transfer therapy of CIK cells has a certain inhibitory effect on tumor cells. However, the difference of tumor volume between hIL-15-IRES-TK plasmid transfected and non-transfected CIK cells was still significant, indicating that hIL-15-IRES-TK plasmid transfected CIK cells had a longer time to inhibit tumor growth than non-transfected CIK cells. During the course of the experiment, two nude mice had tumor ulceration after the IL-15 transfection CIK group began treatment, the second week began to ulcerate scarring, the fourth week ulcerated and healed, the tumor disappeared. There was no such phenomenon in the control group and CIK group. This phenomenon also confirmed that CIK cells transfected with IL-15-IRES-TK plasmid had stronger and more sustained tumor killing effect than non-transfected CIK cells. Conclusion: CIK cells can effectively inhibit the growth of gastric adenocarcinoma cell line BGC-823 in vivo, and hIL-15-IRES-TK plasmid transfected CIK cells can not only inhibit tumor, but also cause tumor regression.
【學(xué)位授予單位】：中國人民解放軍軍醫(yī)進(jìn)修學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2009
【分類號(hào)】：R392
本文編號(hào)：2353540
[Abstract]:In this study, hIL-15-IRES-TK retrovirus vector was constructed, and it was confirmed that it could be transfected into CIK cells effectively. The transfection can make CIK cells secrete interleukin-15 (IL-15). In view of the potential risk of autoimmune disease caused by retrovirus transfection, the expressed thymidine kinase gene (HSV-TK) can effectively remove the transfected CIK cells under the treatment of (GCV). This provides an important basis for the safety of hIL-15-TK virus vector in clinical CIK cell adoptive metastasis. To observe the therapeutic effect of human T cells transfected with hIL15-IRES-TK plasmid on human gastrointestinal carcinoma in vivo. We successfully established a tumor-bearing mouse model of human gastric cancer cells by using T cell immunodeficient mice (Balb/c-nu/nu mice) with human poorly differentiated gastric adenocarcinoma cell line BGC-823. HIL-15-IRES-TK retrovirus-transfected and untransfected CIK cells were injected locally into the tumor once a week for 2 weeks. Saline injection was used as the blank control group. The tumor volume of nude mice was measured once a week. After 4 weeks, the nude mice were killed and the tumor tissue was weighed. After two weeks of treatment, there was a significant difference in tumor volume between the two groups injected with hIL-15-IRES-TK plasmid and non-transfected CIK cells, respectively. The results showed that CIK cells transfected with hIL-15-IRES-TK plasmid had stronger inhibitory activity against tumor growth than non-transfected CIK cells. At the 3rd week, the tumor volume of the three groups increased after CIK treatment, but the hIL-15-IRES-TK plasmid transfection and non-transfection CIK cells were smaller than the normal saline group. These results suggest that adoptive transfer therapy of CIK cells has a certain inhibitory effect on tumor cells. However, the difference of tumor volume between hIL-15-IRES-TK plasmid transfected and non-transfected CIK cells was still significant, indicating that hIL-15-IRES-TK plasmid transfected CIK cells had a longer time to inhibit tumor growth than non-transfected CIK cells. During the course of the experiment, two nude mice had tumor ulceration after the IL-15 transfection CIK group began treatment, the second week began to ulcerate scarring, the fourth week ulcerated and healed, the tumor disappeared. There was no such phenomenon in the control group and CIK group. This phenomenon also confirmed that CIK cells transfected with IL-15-IRES-TK plasmid had stronger and more sustained tumor killing effect than non-transfected CIK cells. Conclusion: CIK cells can effectively inhibit the growth of gastric adenocarcinoma cell line BGC-823 in vivo, and hIL-15-IRES-TK plasmid transfected CIK cells can not only inhibit tumor, but also cause tumor regression.
【學(xué)位授予單位】：中國人民解放軍軍醫(yī)進(jìn)修學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2009
【分類號(hào)】：R392
【引證文獻(xiàn)】
相關(guān)碩士學(xué)位論文 前1條
1 甘正藝;烏司他丁對(duì)大鼠光氣所致急性肺損傷的抑制及對(duì)IL-15和ICAM-1的影響[D];復(fù)旦大學(xué);2012年
，本文編號(hào)：2353540
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