摘要
目的 建立稳定表达核蛋白(NP)的P815细胞株,并以此作为CTL的靶细胞,研究汉滩病毒(HTHV)诱导产生的NP抗原特异性CTL。方法 脂质体介导pEFBOS/HTHVNP和pDORneo质粒DNA共转染P815细胞株,用免疫组化染色法证实NP在胞浆中的表达。分别取HTHV感染小鼠的脾细胞和pEFBOS/HTHVNP质粒DNA免疫小鼠的脾细胞,经体外抗原和IL2再刺激诱导产生CTL效应细胞,进行4h51Cr释放实验。结果 杀伤试验结果表明,无论通过病毒感染还是基因免疫获得的CTL效应细胞,均可以特异性杀伤NP基因转染的靶细胞。结论 HTHVNP可以有效诱导CTL的产生,该研究结果为进一步设计HTHV合成肽疫苗或基因疫苗提供了重要实验依据。
Objective To investigate whether HTHV NP could induce specific CTL recognize NP. Methods A cell line P815 which was co transfected with pEF BOS/HTHV NP and pDOR neo by lipofectin mediation was established. Expression of NP in the cytoplasm of transfected cells was confirmed by immunohistochemistry staining. After restimulation in vitro with HTHV plus rhuIL 2, the splenocytes either from HTHV infected mice or from pEF BOS/HTHV NP plasmid DNA immunized mice were used as effectors in 4?h 51 Cr release assay. Results CTL induced not only by viral infection but also by gene immunization could specifically lyse NP expressed P815 cells. Conclusion Our results suggest that NP of HTHV can efficiently induce CTL which provide an experimental basis for the future design of efficient synthetic peptide vaccine or gene vaccine for HFRS.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2000年第1期23-25,共3页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金!3 970 0 12 8
