摘要
目的 研究WT1多肽疫苗对荷人单核细胞白血病重度联合免疫缺陷(SCID)小鼠的抗肿瘤免疫效应。方法 对SCID小鼠腹腔注射人外周血淋巴细胞(PBL)后24h,皮下接种THP1细胞,建立SCID小鼠荷人单核细胞白血病模型,随机分组,每组8只。空白对照组的疫苗成分为不完全弗氏佐剂,辅助T细胞表位组的疫苗成分为辅助T细胞表位和不完全弗氏佐剂,WT1多肽组的疫苗成分为WT1多肽、辅助T细胞表位和不完全弗氏佐剂。当肿瘤体积约为100mm3时,开始腹腔注射疫苗成分,疫苗接种14d后处死SCID小鼠。采用乳酸脱氢酶法检测小鼠脾细胞特异性CTL杀伤活性,肿瘤组织HE染色后镜下观察组织学特点,利用流式细胞仪检测外周血CD3^+/CD4^+T细胞、CD3^+/CD8^+T细胞及CD4^+CD25^+T细胞水平,应用ELISA方法检测血清中免疫球蛋白、IL-2、IFN-γ、TGF-β及IL-10水平。结果 实验成功建立荷人单核细胞白血病SCID小鼠模型,各组中所有动物均成瘤;WT1组小鼠脾细胞对THP1细胞的CTL活性高于辅助T细胞表位组和空白对照组(P<0.05);WT1组小鼠的肿瘤平均体质量及体积均明显低于辅助T细胞表位组和空白对照组(P<0.05);WT1组肿瘤组织HE染色显示肿瘤细胞变性坏死,存活的肿瘤细胞减少;WT1组小鼠外周血CD3^+/CD4^+T细胞、CD3^+/CD8^+T细胞、IgG、IFN-γ和IL-2水平均明显高于辅助T细胞表位组和空白对照组(P<0.05);外周血CD4^+/CD25^+Treg细胞、TGF-β和IL-10水平均明显低于辅助T细胞表位组和空白对照组(P<0.05)。结论 WT1多肽疫苗可在荷人单核细胞白血病SCID小鼠体内产生抗肿瘤免疫效应,有效杀伤白血病细胞。
Objective To study the anti-tumor immune effects of WT1 peptide vaccine in SCID mice with xenografted human monocytic leukemia.Methods Twenty-four hours after intraperitoneal injection of human peripheral blood lymphocytes, the xenograft human monocytic leukemia model in SCID mice was established by subcutaneous inoculation of THP1 cells.The mice were randomly divided into three groups with eight mice in each.Blank control group was vaccinated with incomplete Freund s adjuvant (IAF).Helper T cell epitope group was vaccinated with helper T cell epitopes and IAF.WT1 group was vaccinated with WT1 peptide, helper T cell epitopes and IAF.When the tumor volume grew to 100 mm 3, intraperitoneal injection of vaccine components started.The SCID mice were killed 14 days after vaccination.LDH release method was adopted to detect the specific CTL killing activity of spleen cells.Histological characteristics of tumor tissue were observed under microscope after HE staining.Flow cytometry was used to test the levels of peripheral blood CD3^+/CD4^+T cells, CD3^+/CD8^+T cells and CD4 +CD25 + Treg cells.ELISA method was applied to detect the levels of serum immunoglobulin, IL-2,γ-interferon, TGF-β and IL-10.Results The xenograft human monocytic leukemia model was successfully established in SCID mice and tumor developed in all the SCID mice.In WT1 group, the activity of mouse spleen cells on THP1 cells was significantly higher than that in helper T cell epitope group and control group ( P< 0.05).The mean weight and volume of tumor were significantly lower in WT1 group than in helper T cell epitope group and control group ( P< 0.05).A large amount of tumor cell degeneration and necrosis was observed under the microscope in WT1 group mice and few tumor cells survived.Peripheral blood levels of CD3^+/CD4^+T cells, CD3^+/CD8^+T cells, IgG, IFN-γ and IL-2 were all higher in WT1 group than in helper T cell epitope group and control group ( P <0.05).However, peripheral blood levels of CD4^+/CD25^+Treg cells, TGF-β and IL-10 were all
作者
李静
张王刚
钟波
白菊
刘海燕
王慧渊
耿妍
LI Jing;ZHANG Wang-gang;ZHONG Bo;BAI Ju;LIU Hai-yan;WANG Hui-yuan;GENG Yan(Department of Pediatrics, The Second Affiliated Hospital of Xi an Jiaotong University, Xi an 710004, China;Department of Hematology, The Second Affiliated Hospital of Xi an Jiaotong University, Xi an 710004, China)
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2019年第5期690-695,共6页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.82170596)
陕西省重点研发计划项目(2018SF-076)~~
