转化生长因子β不敏感的树突状细胞疫苗对肾癌的免疫治疗作用

Immunotherapeutic efficacy of transforming growth factor-beta insensitive dendritic cell vaccine in mice with renal carcinoma

目的观察转化生长因子β（TGF-β）不敏感、肿瘤细胞裂解物负载的树突状细胞（DC）疫苗对小鼠肾癌的治疗作用。方法利用修饰的TβRII粒构建逆转录病毒载体，转染肾癌Renca细胞裂解物负载的DC，获得表达显性负相TpRII（TpRIIDN）的DC，流式细胞仪测定细胞表型变化，观察TGF-β体外增殖抑制效果，Western blot检测SMAD-2的磷酸化水平。Renea细胞皮下荷瘤BALB／C鼠40只，随机分TβRIIN—DC组、DC-抗原组、DC组和对照组4组，每组10只，分别给予不同的皮下接种，观察肿瘤体积改变和小鼠生存期。结果经方差分析，TβRIIDN—DC组与DC-抗原组，DC、组的DC细胞CD86、CD80，CD40、CD11c及MHC-Ⅱ表型差异无统计学意义（P＞0．05），TGF-β对TβRIIDN—DC细胞的增殖无抑制作用。在TβRIIDN—DC组未检测到磷酸化SMAD-2。TβRIIDN—DC组、DC抗原组、DC组和对照组的小鼠肿瘤体积分别为（36．27±13．09）、（115．51±20．75）、（218．10±18．93）和（239．21±21．82）mm^3，4组荷瘤小鼠生存期分别为（75．6±10．2）、（44．3±8．5）、（19．0±5．4）和（21．2±6．7）d，其中TβRIIDN—DC组2只小鼠肿瘤完全消失，TβRIIDN-DC组肿瘤体积及荷瘤小鼠生存期与其他各组比较差异均有统计学意义（P＜0．01）。结论TGF-β不敏感的DC疫苗对肾癌荷瘤BALB／C小鼠有明显的免疫治疗作用。

Objective To investigate the therapeutic effects of dendritic cells （DCs） with transforming growth factor-beta （TGF-β） insensitiveness and acquiring tumor lysate antigen on renal cell carcinoma. Methods DCs were pulsed with freeze-thawed tumor lysate. Then the DCs were rendered TGF-β insensitive by infected with a retrovirus containing dominant-negative TGF-β type II receptor （TβRIIDN）. The immunophenotype of TGF-β-insensitive DCs were analyzed by flow cytometry. The DCs were incubated with TGF-β1 to observe the antiproliferative effects of TGF-and detect phosplaolated SMAD-2. Forty BALB/C tumor-bearing mice were divided into four groups randomly, which were TβRIIDN-DC treatment,DC-antigen treatment,DC treatment and control. Different subcutaneous inoculation was given to those groups. Gross tumor volume and survival time of tumor-bearing mice viere observed. Results By analysis of variance, difference of the phenotype CD86 、CD80 ,CD40, CD11c and MHC-Ⅱ was not statistically significant among TβRIIDN-DC group,DC-antigen group and DC group（P〉0.05）. TGF-β-insensitive DCs were resistant to the antiproliferative effects of TGF-β. Phosphorylated SMAD-2 was undetectable in TGF-β-insensitive DCs. Gross tumor volume of tumor bearing mice in TβRIIDN-DC treatment. DC-antigen treatment. DC treatment and control was（36.27±13.09）mm^3 .（115.51±20.75）mm^3 .（218.10±18.93 ）mm^3 and （239.21±21.82）mm^3 , respectively. Survival time of tumor-bearing mice was （75.6±10.2）d. （44.3±8.5）d. （19.0±5.4）d and（21.2±6.7） d, respectively. Furthermore, complete tumor regression occurred in 2 vaccinated mice of TβRIIDN-DC group. Difference of gross tumor volume and survival time of tumor-bearing mice was statistically significant between TβRIIDN-DC group and other groups（P〈0.01） o Conclusions The TGF-β-insensitive DC vaccine can induce the specific and effective immune response against renal carcinoma in BALB/C mice.