摘要
目的 通过体外培养的Th17细胞过继免疫治疗弥漫大B细胞淋巴瘤（DLBCL）小鼠,分析白细胞介素17（IL-17）水平与主要组织相容性复合体Ⅱ（MHCⅡ）表达的相关性及其与肿瘤发生发展的关系.方法 免疫磁珠法分离纯化BALB/c小鼠脾来源的CD4＋CD62L＋T细胞,加入抗CD3抗体、抗CD28抗体、转化生长因子β（TGF-β）、IL-6体外培养Th17细胞,人生发中心B细胞样（GCB）DLBCL细胞株SUDHL-4传代培养后接种到严重联合免疫缺陷（SCID）小鼠,建立DLBCL小鼠模型.将荷瘤小鼠分为Th17细胞实验组（30只）和对照组（20只）,实验组荷瘤小鼠接种Th17细胞,对照组注射0.9％NaCl溶液.分别于预实验得到的小鼠肿瘤中位发病时间和小鼠中位生存时间处死半数小鼠.酶联免疫吸附法（ELISA）检测小鼠肿瘤组织IL-17表达,免疫组织化学法检测MHCⅡ的表达.结果 DLBCL小鼠肿瘤中位发病时间为8d,中位生存期为28d.Th17细胞接种后IL-17表达水平[（11.93±0.56）pg/ml]较对照组[（9.82±0.26）pg/ml]升高（P＜ 0.000 1）,随着肿瘤病程进展,IL-17表达水平降低[（9.53±0.18）pg/ml]（P＜ 0.000 1）;Th17细胞接种后MHCⅡ阳性细胞比例[（69.13±0.36）％]较对照组[（42.59±0.12）％]升高（P＜ 0.000 1）,随着肿瘤病程进展,MHCⅡ阳性细胞比例降低[（54.63±0.45）％]（P＜ 0.000 1）.IL-17与MHCⅡ之间存在正相关性（r=0.89,P=0.000）.结论 IL-17可上调DLBCL小鼠肿瘤组织MHCⅡ表达,且随着肿瘤病程进展,MHCⅡ表达下降.MHCⅡ表达水平可作为DLBCL疾病状态的判断指标,而其正向调控因子IL-17表达水平的上调会影响DLBCL的疾病进程,联合检测IL-17和MHCⅡ表达水平对判断DLBCL疾病状态及进程可能更有参考价值.
Objective To establish a mice model of diffuse large B-cell lymphoma （DLBCL） that was treated with adoptive immunity of Th17 cells cultured in vitro,and to analyze the relationship between IL-17 and MHC Ⅱ expression and their relation with tumor growth.Methods The CD4＋CD62L＋ T cells purified by MACS were stimulated under cytokine conditions including anti-CD3,anti-CD28,TGF-β and IL-6 in vitro,and SUDHL-4 cells were cultured and inoculated the SCID mice to establish DLBCL mice models.The mice were divided into Th17 cells immunity group （30 mice） and control group （20 mice）.Th17 cells were injected to mice to get the adoptive immunity in immunity group,and 0.9 % NaCl in control group.The half mice were terminated at median disease onset time and median survival time,respectively.ELISA was used to detect IL-17 expression,and immunohistochemistry was applied to detect MHC Ⅱ expression in the tumor tissues.Results The median disease onset time of DLBCL mice model was 8 d,and median survival time was 28 d.The IL-17 and MHC Ⅱ expression levels in Th17 cells immunity group [（11.93±0.56） pg/ml,（69.13t0.36） %] were higher than those in control group [（9.82±0.26） pg/ml,（42.59±0.12） %] （both P〈 0.000 1）.Along with the progress of DLBCL,IL-17 and MHC Ⅱ expression levels were decreased [（9.53±0.18） pg/ml,（54.63±0.45） %,both P 〈 0.000 1].There was a significantly positive correlation between IL-17 and MHC Ⅱ （r=0.89,P=0.000）.Conclusions The expression level of MHC Ⅱ can be used as a factor to judge the disease situation of DLBCL,and combination detection of the expression of both IL-17 and MHC Ⅱ will provide more reference values for judgment of the disease situation and the progress of DLBCL.
出处
《白血病．淋巴瘤》
CAS
2016年第8期461-464,470,共5页
Journal of Leukemia & Lymphoma
基金
广东省科技计划（20138021800040）
