摘要
目的将鼠源肾母细胞瘤WT1基因一段序列(WT1y)构建于真核表达质粒pCDNA3.1(+),通过小鼠和体外细胞模型初步研究所激发的针对肾母细胞瘤CTL效应。方法人工合成WT1基因一段序列,含有HLA-A*2402锚定残基的9个氨基酸。构建重组真核表达质粒pCDNA3.1(+)/WT1y。免疫Balb/c小鼠,通过ELISA方法检测体液免疫反应;分离脾淋巴细胞,FCM检测脾淋巴细胞中CD4/CD8;将分离的脾淋巴细胞与小鼠肾母细胞瘤细胞共培养,检测其体外溶解组织相容性抗原型别一致的外源性靶细胞能力。结果构建的重组质粒经测序鉴定,与GenBank中登录的序列完全一致;免疫组小鼠T淋巴细胞增殖及CTL活性均明显优于对照组(P<0.01);体外具有较强溶解靶细胞的功能。结论WT1基因的DNA疫苗初步研究具有很强的CTL效应,为小儿肾母细胞瘤的治疗提供了新的思路。
Objective To construct the eukaryotie expression vector pCDNA3.1 （ ＋ ） containing WT1 y gene an assay the specific CTL responses to DNA vaccine of Wilms＇ tmnor gene （WT1y） product. Methods The WT1y gene of 321 bp, including HLA-A ＊ 2402 anchoring residue; was synthetized, and then transfected into eukaryotic vector calledpCDNA3.1 （ ＋ ）. The constructed eukaryotic vector calledpCDNA3.1（ ＋ ）/WT1y was used to immunize Balb/c mice. WT1-specific antibody was detected in serum of mice by ELISA method. T lymphocyte proliferation was detected by FCM. The CTLs response was detected by co-culture the murine spleen cells and the tumor cells. Results Recombinant gene was confimaed using DNA sequencing. WT1-specific antibodies were detected in serum of immunized mice by ELISA. T lymphocyte proliferation of immunized mice was superior to that of control group （ P 〈 0.01）.The CTLs exerted specific lysis against WT1-expressingcells. Conclusion The DNA vaccine of WT1y can induce specific CTLs , which offers a new way for treatment of Wilms＇ tumor.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第3期252-254,259,共4页
Immunological Journal
基金
国家自然科学基金资助项目(30170853)
