目的比较DC-LMP2与rAd—LMP2在BALB/c小鼠中的免疫效果。方法从BALB/c小鼠中分离骨髓细胞,在细胞因子作用下体外诱导培养获得小鼠树突状细胞(mDC),再以100MOI的含EB病潜伏膜蛋白2的重组腺病毒(rAd—LNP2)感染,获得DE—LNP2,免疫酶法确定LMP2在DC中表达。于0,2,4周,分别以PBS、DC-LMP2和rAd-LMP2肌肉免疫BALB/c小鼠,第5和8周检测小鼠体内LMP2特异性细胞应答水平。结果骨髓细胞体外成功诱导获得DC,LMP2在DC中表达。DC—LMP2和rAd—LMP2均可诱导LMP2特异性细胞免疫应答,随时间推移而减弱。rAd—LMP2诱导特异性细胞免疫应答水平高于DC—LMP2。结论肌肉免疫rAd—LMP2能够在BALB/c小鼠中诱导较强的LMP2特异性细胞免疫应答。
Objective To compare immunological effect induced with DC-LMP2 and rAd-LMP2 in BALB/c mice. Mehtods Bone marrow cells were separated from BALB/c mice, and induced into murine dendritic ceils (mDCs) cultured by RPMI 1640 medium supplemented with 10% fetal calf serum and recombinant murine cytokines in vitro, mDCs were infected by recombinant adenovirus including latent membrane protein l] (LMP2) gent (rAd-LMP2) of Epstein-Barr virus(EBV) at a dose of 100 multiplicity of infection (MOI) , and matured mDC-LMP2 was harvested at 48 hours later. The LMP2 expression in mDCs was detected by immunoenzyme. BALB/c mice were immunized with PBS DC-LMP2 and rAd-LMP2 by intramuscular injection at 0,2,4 weeks. LMP2 specific cytotoxic lymphocytes (CTLs) in BALB/c mice were detected at 5th week and 8th week with enzyme-linked immunosorbent spot (ELISPOT). Results mDCs from bone marrow cells were harvested. LMP2 was detected in DC-LMP2. LMP2 specific CTLs were stimulated by both DC-LMP2 and rAd-LMP2 and decreased with time elapsed. LMP2 specific CTLs stimulated by rAd-LMP2 were higher than that by DC-LMP2 in BALB/c mice. Conclusion Immunization with rAd-LMP2 by intramuscular injection was more fitful than that with DC-LMP2 in BALB/c mice.