摘要
目的 观察影响CTL体外杀伤效应的特异性和敏感性的主要因素。方法 抗原OVA免疫小鼠C5 7BL 6 ,取免疫后小鼠脾细胞 ,先用ELISPOT法检测CTL的活化 ,然后抗原肽OVA2 57- 2 6 4体外刺激免疫后的脾细胞 5d ,51 Cr释放法观察不同因素对CTL的体外杀伤效果的影响。结果 ELISPOT检测结果显示抗原免疫组产生了强CTL应答 ,而PBS免疫组没有应答。游离多肽的存在显著降低杀伤的效率 ,IL 2和Ficoll Hypaque离心去除死细胞可提高体外杀伤的效率。ConA的存在可以使CTL体外非特异的杀伤靶细胞。结论 CTL体外杀伤实验中必需尽量去除游离多肽 。
Objective To explore the factors that affect the specificity and sensitivity of CTL killing in vitro. Methods The activation of mouse spleen cells immunized by antigen OVA was determined by ELISPOT assay. To further explore the effect of various factors on the CTL killing efficiency, immunized spleen cells were stimulated with antigen peptide OVA 257-264 in vitro for 5 d. CTL killing ability and specificity was detemined by standard 51Cr release assay. Results Strong CTL response was observed in immunized mouse whereas no response was detected in PBS control mouse. The CTL killing efficiency decreased dramatically in the presence of free pepti des, but the addition of exogenous rIL-2 and removal of dead cells from culture by Ficoll-Hypaque density gradient centrifugation could increase the CTL killing ability. Spleen cells could kill target cells nonspecifically in vitro in the presence of mitogen ConA. Conclusion To get high specificity and sensitivity in the experiment of CTL killing in vitro, free peptides should be removed as complete as possible.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2004年第3期169-171,176,共4页
Immunological Journal
基金
国家自然科学基金 (30 2 0 0 2 51 )
海外杰出青年基金资助项目 (30 1 2 80 1 9)
