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目的研究 GM-CSF 作为免疫佐剂对胰腺癌 MUC1-VNTR 核酸疫苗免疫效果的增强作用。方法 45只 C57BL/6小鼠随机分3组,在胫前肌注射100μg/100μl质粒 DNA 生理盐水溶液(VG 组接种 pcDNA3.1-VNTR/Myc-his(+)A 质粒联用 rmGM-CSF,V 组单纯接种 pcDNA3.1-VNTR/Myc-his(+)A 质粒,G 组单纯接种 rmGM-CSF)进行免疫接种。4周后加强免疫1次,6周后取血清 ELISA 法测抗体,取脾细胞悬液体外以 VNTR 合成肽特异刺激,6d 后进行杀伤试验。结果VG 组小鼠脾细胞 CTL 的特异性杀伤率显著高于 V 组(P<0.01)和 G 组(P<0.01),表明 GM-CSF增强了疫苗所诱发的 CTL 应答。而 CTL 对未经 VNTR 合成肽孵育的靶细胞 EL4-VNTR~-杀伤较低(P<0.01),VU 3C6可抑制 CTL 对经 VNTR 合成肽孵育的靶细胞 EL4-VNTR~+的杀伤活性(P<0.01),表明具有 VG 组诱生的 CTL 应答依然保持 VNTR 特异性。VG 组小鼠血清抗 VNTR 抗体等效浓度(3119.0±110.6)μg/ml 高于Ⅴ组(2323.5±238.3)μg/ml 和 G 组(2023.9±169.0)μg/ml,差别极为显著(P<0.01)。结论 GM-CSF 可增强自行构建的胰腺癌 MUCl-VNTR 核酸疫苗所诱生的 VNTR 特异的 CTL 应答和抗体应答。
Objective To study the enhancing effect of GM-CSF as immune adjuvant on the immune effect of MUC1-VNTR DNA vaccine against pancreatic cancer. Methods Forty five C57BL / 6 mice were randomly divided into three groups: 100μg / 100μl of plasmid DNA solution was injected into the anterior tibial muscle (VG group was inoculated with pcDNA3.1-VNTR / Myc-his (+) A plasmid in combination with rmGM- Group V was inoculated with pcDNA3.1-VNTR / Myc-his (+) A plasmid and Group G was inoculated with rmGM-CSF alone). After 4 weeks, the immune system was boosted once. Serum ELISA was used to measure the antibody after 6 weeks. The spleen cell suspension was stimulated with VNTR synthetic peptide in vitro and killed after 6 days. Results The specific cytotoxicity of CTL in splenocytes in VG group was significantly higher than that in V group (P <0.01) and G group (P <0.01), indicating that GM-CSF enhanced the CTL response induced by the vaccine. However, the cytotoxicity of CTL on EL4-VNTR ~ + cells incubated with VNTR synthetic peptide was lower (P <0.01) and VU 3C6 inhibited the cytotoxic activity of CTL on EL4-VNTR + <0.01), indicating that the CTL response induced by the VG group still retains the VNTR specificity. The mean concentration of anti-VNTR antibody (3119.0 ± 110.6) μg / ml in VG group was significantly higher than that in group V (2323.5 ± 238.3) μg / ml and G group (2023.9 ± 169.0) μg / ml ). Conclusion GM-CSF can enhance the VNTR-specific CTL response and antibody response induced by self-constructed pancreatic cancer MUC1-VNTR DNA vaccine.