摘要
目的 研究肿瘤抗原p185蛋白作为肿瘤疫苗所诱导的抗肿瘤效应。方法 亲和层析纯化的p185蛋白皮下注射正常BALB c小鼠 ,取脾细胞分别与p185蛋白、3T3 neu细胞体外共培养 ,以单独佐剂注射组的小鼠脾细胞为对照 ,观察其培养上清中IFN γ和TNF α活性。将免疫鼠脾淋巴细胞经p185抗原和低剂量的IL 2体外培养 1周后 ,再用CD3抗体刺激扩增 ,用流式细胞仪分析其细胞表型。 3T3 neu和 3T3细胞分别接种于裸鼠腋窝皮下验证其致瘤性。选择具有致瘤性的 3T3 neu细胞与经抗CD3抗体扩增的免疫鼠脾细胞体外混合后 (1 9)接种于裸鼠腋窝皮下 ,以 3T3 neu细胞单独或与CD3抗体活化的正常鼠脾细胞 (CD3 AK)混合后 ,同法接种于裸鼠腋窝皮下观察其成瘤时间和带瘤宿主的存活期。结果 免疫鼠脾细胞培养上清中IFN γ活性为 40 .2 7± 9.6U ml,也具有产生较高TNF α能力。对照组未能检测出有IFN γ活性 ,其TNF α活性也较低。裸鼠体内实验证实 ,2 .5× 10 5的 3T3 neu细胞在裸鼠体内即可成瘤 ,而对照组的 3T3细胞无致瘤性。 3T3 neu细胞与体外扩增的免疫鼠脾细胞混合后接种裸鼠 ,瘤结节形成时间平均为 5 2 .5d ,存活期平均为 5 5 .3d(其中 5只动物至6 0d尚未成瘤 ,按 6 0d计算 ) ,而对照组瘤结节形成时间平均分别为 12 .5和 9.
Objective To study the anti tumor effect of p185 oncoprotein as a tumor vaccine. Methods The anti tumor effect in vitro/vivo was studied by using the p185 antigen as a tumor protein vaccine to immunize mice. The splenic lymphocytes from the immunized mice were incubated with p185 tumor antigen for 4 days in vitro, then the supernatant was harvested for detecting the IFN γ activity. The immune splenic lymphocytes in presence of a low dose(10μg/ml)IL 2 containing medium were cultured with p185 antigen for one week and expanded with anti CD3 antibody. The phenotypes of expanded lymphocytes were detected by FACS. The TNF activity secreted by the expanded lymphocyte cells after co incubated with 3T3 neu cells for 4 hours was also tested. 3T3 neu cells with tumorgenicity in nude mice admixed with the expanded lymphocytes (19) in vitro and was then inoculated (s.c.) into nude mice, 3T3 neu cells were inoculated alone or admixed with anti CD3 activated spleen cells (CD3 AK) as control group. Results IFN γ activity in the supernatant was 40.27±9.6U/ml in experimental group(splenocytes from immunized mice), but was undetected in control group (immunized with adjuvant alone). 86.2% expanded lymphocytes were CD3 positive T cells, in which 23% were CD8 positive T cells. The TNF activity was significantly higher in the supernatant from the experimental spleen cells culture than that from control (P<0.001). The mean occurrence time of the tumor nodes was 52.5 days in experimental group (among no tumor occurs in 5/6 animals within more than 60 days after inoculation) and 12.5 days or 9.2 days in both control group respectively. The mean survival time of the tumor bearing nude mice was significantly extended in experimented group (55.3 days) than in control group (27.8 days and 25.6 days) respectively. Conclusion These findings indicated that the p185 specific T cells may have an ability to inhibit the 3T3 neu tumor growth in vivo.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2000年第3期258-261,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目!(39640009)
