摘要
目的:研究ΔNp73α基因转染树突状细胞(DC)诱导的特异性抗乳腺癌免疫效应。方法:人脐带血细胞经GM-CSF、IL-4、TNF-α等细胞因子诱导培养DC,流式细胞仪(FCM)检测DC成熟前后CD1a、CD83的表达变化情况。脂质体法将pc DNA-HA/ΔNp73α转染至DC,经Western blot检测转染情况。转染DC与自体T细胞共培养诱导特异性CTL。MTT法测定T细胞增殖能力;ELISA法检测IFN-γ的分泌水平;LDH释放法检测T细胞对乳腺癌细胞MDA-MB-231的杀伤作用。结果:DC诱导成熟后,CD1a表达约占56%,CD83约占74%,与未成熟DC(CD1a 19%,CD83 13%)比较,差异有显著统计学意义(P<0.01)。Western blot检测到DC-ΔNp73α组有一特异条带表达。DC-ΔNp73α组诱导的特异性CTL对MDA-MB-231杀伤作用高于DC组(P<0.05),而且刺激T细胞增殖能力增强,分泌IFN-γ的水平升高,与空载体DC-pc DNA组及DC组比较有显著统计学意义(P<0.01)。结论:以ΔNp73α转染DC制备的DC疫苗,具有显著诱导CTL杀伤乳腺癌细胞的作用。
Objective：To research the imumunological effect of dendritic cell transfected by recombinant plasmid of ANp73a on breast cancer. Methods： Cultivate dendritic cells by cytokines（ GM-CSF, IL-4, TNF-a） from human umbilical cord blood in vitro, CD1 a,CD83 were detected by flow eytometry. Recombinant plasmid pcDNA-HA/ANp73a was used to transfected DCs,the transfection was detected by Western blot. Transfected DCs cocuhivate with T cell induce specific cytotoxie lymphocytes（CTL）. Proliferation ability of T cell was detected by MTr methods. IFN-＇y in the culture supernatant of the transfected DCs were detected by ELISA. The killing effect on breast cancer MDA-MB-231 was detected by LDH method. Results： The expression of CD1 （56% ）, CD83 （74%）in mature DC was higher than immature DC （ CDla 19% , CD83 13% ） （ P〈0. 01 ）. △Np73a transfected group had a special band. The killing effect of specific CTL induced by △Np73a-DC and T cells cocuhivation on breast cancer cell MDA-MB-231 was statistic stronger than DC group（P〈0.05 ） , the proliferation of T cell and secretion level of IFN-7 compared with DC and pcDNA group were statistically significant （ P〈0. 01 ）. Conclusion： The CTL induced by △Np73a transfected DC vaccine has significant effect on killing breast cancer.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2016年第2期169-173,共5页
Chinese Journal of Immunology
基金
辽宁省自然科学基金资助项目(201102127)
