SCID鼠人肝癌皮下移植及免疫重建复合模型的建立

Establishment of SCID murine model of subcutaneous transplantation of human hepatocellular carcinoma and reconstruction of human immune system

目的 探索建立稳定的SCID鼠人肝癌皮下移植及免疫重建 (人HCC PBL SCID)复合模型的方法。方法 经SCID鼠腹腔内注射人PBL、皮下接种人肝癌细胞 ,定期检测SCID鼠体内人淋巴细胞及其功能、观察皮下成瘤潜伏期及成瘤率。结果 人HCC PBL SCID复合模型鼠和单纯人HCC SCID模型鼠成瘤率均为 10 0 % ,但前者成瘤潜伏期显著延长、肿瘤明显缩小 (P分别 <0 0 1、<0 0 5 )。人HCC PBL SCID复合模型鼠第 2、4、6周时小鼠血中人IgG分别为 (6 9 8± 11 6 ) μg/ml、(12 5 9± 13 7) μg/ml和(183 1± 9 0 ) μg/ml,人CD3+ 淋巴细胞占单核细胞分别为 (10 5± 5 2 ) %、(9 7± 2 9) %和 (8 4± 2 4 ) %。复合模型鼠第 4、6周的小鼠脾细胞对HepG2细胞的杀伤力分别为 (2 0 3± 8 7) %和 (2 2 0± 2 3) %。免疫组化检测显示小鼠脾脏、胸腺、肿瘤组织中均有较多的人CD3+ 淋巴细胞分布。结论 采用腹腔注射人PBL、皮下接种人肝癌细胞的方法可以有效地建立人HCC PBL SCID嵌合模型。此模型较好地模拟了肝癌的人体内情况 ,是肝癌免疫基因治疗的较理想模型。

Objective To investigate the method for establishing stable SCID murine model of subcutaneous transplantation of human hepatocellular carcinoma and reconstruction of human immune system (hu HCC-PBL-SCID). Methods The survival and function of human lymphocytes and latency period and rate of tumor formation in SCID mice were monitored after intra-peritoneal injection of human peripheral blood lymphocytes (PBL) and subcutaneous implantation of human HCC cells. Results Subcutaneous tumors developed in all the mice given human PBL and HCC cells. Compared with mice subject to subcutaneous implantation of HCC cells alone, the latency period in hu HCC-PBL-SCID murine model was significantly prolonged, and the tumor size was markedly depressed (P<0.01, 0.05, respectively). In the 2nd, 4th and 6th week, human IgG in the murine peripheral blood of the hu HCC-PBL-SCID model was 69.8±11.6 μg/ml, 125.9±13.7 μgml and 183.1±9.0 μg/ml and the human CD3+ lymphocytes 10.5±5.2%, 9.7±2.9% and 8.4±2.4%, respectively. The killing efficacy of the murine splenocytes of the hu HCC-PBL-SCID model on the 4th and 6th week to HepG2 cells was 20.3±8.7% and 22.0±2.3%, respectively. Immunohistochemical staining revealed presence of remarkable human CD3+ lymphocytes in the murine spleen, thymus and tumor. Conclusions The hu HCC-PBL-SCID murien model could be successfully established by intra-peritoneal injection of human PBL and subcutaneous implantation of human HCC cells. The model well simulates the real situation of HCC in human being and represents an ideal in vivo model for immune gene therapy of HCC.