Chu Xiao-Yuan, Chen Long-Bang, Zang Jing, Wang Jing-Hua, Zhang Qun, Geng Huai-Cheng
Department of Oncology, Nanjing General Hospital of PLA, Nanjing 210002, Jiangsu Province, China.
World J Gastroenterol. 2005 Feb 7;11(5):760-3. doi: 10.3748/wjg.v11.i5.760.
To investigate the effect of bone marrow-derived monocytes transfected with RNA of CT-26 (a cell line of mouse colon carcinoma) on antitumor immunity.
Mouse bone marrow-derived monocytes were incubated with mouse granulocyte macrophage colony stimulating factor (mGM-CSF) in vitro, and the purity of monocytes was detected by flow cytometry. Total RNA of CT-26 was obtained by TRIzol's process, and monocytes were transfected by TransMessenger in vitro. The activity of cytotoxic T lymphocytes (CTL) in vivo was estimated by the modified lactate dehydrogenase (LDH) release assay. Changes of tumor size in mice and animal's survival time were observed in different groups.
Monocytes from mouse bone marrow were successfully incubated, and the positive rate of CD11b was over 95%. Vaccination of the monocytes transfected with total RNA induced a high level of specific CTL activity in vivo, and made mice resistant to the subsequent challenge of parental tumor cells. In vivo effects induced by monocytes transfected with total RNA were stronger than those induced by monocytes pulsed with tumor cell lysates.
Antigen presenting cells transfected with total RNA of CT-26 can present endogenous? tumor antigens, activate CTL, and effectively induce specific antitumor immunity.
研究用CT-26（小鼠结肠癌细胞系）RNA转染的骨髓来源单核细胞对抗肿瘤免疫的影响。
将小鼠骨髓来源单核细胞与小鼠粒细胞巨噬细胞集落刺激因子（mGM-CSF）在体外孵育，通过流式细胞术检测单核细胞纯度。采用TRIzol法获取CT-26的总RNA，体外利用TransMessenger转染单核细胞。通过改良乳酸脱氢酶（LDH）释放试验评估体内细胞毒性T淋巴细胞（CTL）活性。观察不同组小鼠肿瘤大小变化及动物存活时间。
成功孵育小鼠骨髓来源单核细胞，CD11b阳性率超过95%。用总RNA转染的单核细胞进行疫苗接种可在体内诱导高水平的特异性CTL活性，并使小鼠对后续亲本肿瘤细胞的攻击产生抗性。用总RNA转染的单核细胞诱导的体内效应强于用肿瘤细胞裂解物脉冲处理的单核细胞诱导的效应。
用CT-26总RNA转染的抗原呈递细胞可呈递内源性肿瘤抗原，激活CTL，并有效诱导特异性抗肿瘤免疫。
