RNA干扰供体大鼠库普弗细胞B7分子表达对受体大鼠淋巴细胞激活的影响  

Effect of RNA interference B7 expression of donor rat Kupffer cells on activation of recipient rat lymphocytes

作　　者：李涛[1] 朱继业[1] 柳枫[1] 冷希圣[1] 

机构地区：[1]北京大学人民医院肝胆外科,北京市100044

出　　处：《世界华人消化杂志》2008年第1期39-44,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金;No.30471637~~

摘　　要：目的：观察RNA干扰供体Lewis大鼠库普弗细胞（KC）B7分子表达对受体BN大鼠淋巴细胞增殖和生成IL-2的影响．方法：分离培养供体Lewis大鼠KC，设计大鼠B7分子的干扰片段，构建并鉴定含B7干扰片段的RNA干扰载体Psilencer 3．1H1-Neo-B7，将RNA干扰载体转染供体大鼠的KC，转染后采用RT-PCR方法检测KC上B7分子表达的变化．将转染后的KC分为3组，对照组（A）；空载体组（B）；RNA干扰B7表达组（C）．分离培养受体BN大鼠的淋巴细胞，将以上各组细胞分别与BN大鼠的淋巴细胞进行共培养，采用MTT法检测各组淋巴细胞的增殖情况．采用ELISA方法检测各组培养上清中IL-2的含量．结果：分离培养的供体Lewis大鼠KC得率为5×10^7，活率大于98％．构建的RNA干扰载体经酶切和测序鉴定正确．RNA干扰KC后其B7的表达降低了22％（P〈0．01）．将干扰B7表达的KC与BN大鼠的淋巴细胞进行共培养，与对照组相比，受体BN大鼠的淋巴细胞增殖降低了49％（P〈0．01），细胞培养上清中IL-2的分泌量下降了67％（P〈0．01）．结论：RNA干扰供体Lewis大鼠KC B7分子的表达可明显抑制受体BN大鼠淋巴细胞的增殖和IL-2的产生．AIM： To investigate the effect of RNA interference B7 expression of donor Lewis rat Kupffer cells （KC） on proliferation and interleukin （IL）-2 production of recipient BN rat lymphocytes. METHODS： Lewis rat KC were isolated and identified. Rat B7 RNA interference vector Psilencer 3.1 H1-Neo-B7 was constructed and identiffed. KC were transfected by B7 RNA interference vector. RT-PCR was used to measure the expression of B7 in KC. After transfection, KC were divided into three groups： group A served as a control group; group B was KC transfected with vector without B7 interference; group C was RNA interference B7 expression of KC. Recipient BN rat lymphocytes were isolated and cocultured with the three groups of KC. MTT assay was used to evaluate lymphocyte proliferation.ELISA was used to detect the production of IL-2. RESULTS： The yield of KC was 5×10^7 and cell viability was 〉 98%： After B7 RNA interference vector transfection, B7 expression of KC decreased by 22% （P 〈 0.01）. After co-culture with recipient BN rat lymphocytes, the proliferation and IL-2 production of lymphocytes were inhibited. As compared with the controls, lymphocyte proliferation decreased by 49% （P 〈 0.01）. IL-2 production decreased by 67% （P 〈 0.01）. CONCLUSION： RNA interference B7 expression of donor Lewis rat KC significantly decreases the proliferation and production of recipient BN rat lymphocytes.

关 键 词：库普弗细胞 B7 共刺激通路 RNA干扰 免疫耐受 

分 类 号：R392[医药卫生—免疫学]