肝癌小鼠脾脏免疫细胞的变化及其意义

The dynamic alteration of splenic immune cells and its significance in hepatoma mice

目的:观察小鼠H22细胞原位种植性肝癌模型中,脾脏正性免疫细胞和负性免疫细胞比例的变化,并且观察脾脏切除后外周血和肿瘤组织免疫细胞比例的变化以及对肿瘤生长的影响。方法:建立小鼠H22细胞原位种植性肝癌模型,在荷瘤的不同时期,用流式细胞术检测脾脏中负性免疫细胞(MDSC、Treg)和正性免疫细胞(总CD3^+T细胞、CD4^+T细胞、CD8^+T细胞、NK细胞、NKT细胞)比例的变化。荷瘤1 w后切除脾脏,用流式细胞术检测外周血及肿瘤组织中免疫细胞比例的变化,并比较切脾前后肿瘤重量、腹水量、荷瘤小鼠生存期的变化。结果:荷瘤小鼠脾脏MDSC细胞的比例一直高于正常组;Treg细胞在荷瘤2 w时显著升高;总CD3^+T细胞和CD4^+T细胞在荷瘤1 w时显著升高,2 w时显著下降;CD8^+T细胞在荷瘤2 w时明显下降;NK细胞在荷瘤3 w时明显下降;NKT细胞无显著变化。脾脏切除后外周血和肿瘤组织MDSC的比例下降,CD8^+T细胞的比例升高;肿瘤重量和荷瘤小鼠生存期无显著变化,腹水量在荷瘤2 w时显著减少,腹水发生率明显降低。结论:小鼠肝脏种植H22细胞株后,脾脏中正性免疫细胞的比例下降,负性免疫细胞的比例升高,脾脏的负性免疫状态逐渐占主导地位,从而促进了肝癌的发展。

Objective：To observe the percentage changes of positive and negative inmmne cells in spleen, and to detect the alteration of the immune status in peripheral blood and tumor tissue after splenectomy. Meanwhile, the tumor growth was compared in splenectomy group and control group in a murine H22 orthotopic hepatoma model. Methods ： The murine H22 orthotopic hepatoma model was established. During the different stage of tumor growth, the percenta- ges of total CD3 ＋ T cells, CD4 ＋ T cells, CD8 ＋ T cells, NK cells and NKT cells in spleen were examined by flow cytom- etry. Splenectomy was performed 1 w after tumor inoculation, then the percentage alterations of immunocytes in periph- eral blood and tumor tissue were detected by flow cytometry. The tumor weight,the amount of ascites and the survival of tumor - bearing mice were also compared after splenectomy. Results ： Compared to normal mice, splenic MDSCs were progressively higher in TB mice. Treg significantly increased on week 2. The total CD3 ＋ T cells and CIM ＋ T cells were elevated on week 1, then declined on week 2. CD8 ＋T cells reduced on week 2,and NK cells decreased on week 3. There was no any- change in the percentage of NKT cells. After splenectomy,the percentage of MDSCs reduced and that of CD8 ＋ T cells increased in peripheral blood and tumor tissue. The tumor weight and the survival time of TB mice had no changes ,whereas, the amount and the incidence of ascites significantly reduced after splenectomy. Con- clusion ： With the development of routine H22 orthotopic hepatoma, the percentages of positive immune cells in spleen reduced and that of negative immune cells in spleen increased, the negative immune status progressively dominated in the spleen, thus facilitated hepatoma growth.