Wang Xue-feng, Zhang Rong-bo, Hu You-ying, Du Jiu-wei, Chen Xiao-jun, Xu Zhi-peng, Su Chuan
Department of Pathogen Biology and Immunology, School of Medicine, Anhui University of Science and Technology, Huainan 232001, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2011 Aug;29(4):319-21.
A C-T-B PDDV mixture of the three constructed epitope-based peptide-DNA dual vaccines (PDDV) containing the CTL (C), Th (T) and B-cell (B) epitopes from Sj22.6 tegument (C-PDDV, T-PDDV and B-PDDV) with a 1:1:1 ratio was prepared. Thirty-six mice were randomly divided into six groups averagely named as 18K group, PBS group, C-PDDV group, T-PDDV group, B-PDDV group, and C-T-B PDDV group. All the mice received three immunizations at 2-week intervals with the same dose of antigen (10 microg DNA+28 microg peptide). One week after the last immunization, the mice were sacrificed, the spleens were removed and splenocytes were collected. Splenocyte proliferation was assayed by[3H] TdR incorporation after stimulation with soluble worm antigen (SWA). Levels of IFN-gamma and IL-4 in the splenocyte culture supernatants were determined by ELISA. The results showed that IFN-gamma content in T-PDDV group [(76.0 +/- 11.2) pg/ml] was higher than that of PBS [(13.0 +/- 2.1) pg/ml] and 18K control groups [(14.0 +/- 3.2) pg/ml] (P<0.01). IL-4 level in T-PDDV [(152.0 +/- 21.1) pg/ml] and C-T-B mixture groups [(86.0 +/- 12.2) pg/ml] was higher than others (P<0.01 and P<0.05). The splenocytes from T-PDDV group showed a significant increase in proliferation compared with PBS and 18K control groups after stimulation by SWA (P<0.01). However, there was no significant difference in splenocyte proliferation among C-T-B, PBS and 18K control groups (P>0.05). These findings indicate that T-PDDV and C-T-B PDDV mixture induces stronger immune response than that of C-PDDV or B-PDDV.
制备了一种由三种构建的基于表位的肽 - DNA 双疫苗(PDDV)组成的 C - T - B PDDV 混合物,其包含来自日本血吸虫 22.6 皮层的细胞毒性 T 淋巴细胞(CTL,C)、辅助性 T 细胞(Th,T)和 B 细胞(B)表位,比例为 1:1:1(C - PDDV、T - PDDV 和 B - PDDV)。36 只小鼠被随机平均分为六组,分别命名为 18K 组、PBS 组、C - PDDV 组、T - PDDV 组、B - PDDV 组和 C - T - B PDDV 组。所有小鼠每隔 2 周接受三次相同剂量抗原(10μg DNA + 28μg 肽)的免疫接种。最后一次免疫接种后 1 周,处死小鼠,取出脾脏并收集脾细胞。用可溶性虫抗原(SWA)刺激后,通过[³H]TdR 掺入法检测脾细胞增殖。通过酶联免疫吸附测定法(ELISA)测定脾细胞培养上清液中 IFN -γ 和 IL -4 的水平。结果显示,T - PDDV 组的 IFN -γ 含量[(76.0±11.2)pg/ml]高于 PBS 组[(13.0±2.1)pg/ml]和 18K 对照组[(14.0±3.2)pg/ml](P<0.01)。T - PDDV 组[(152.0±21.1)pg/ml]和 C - T - B 混合物组[(86.0±12.2)pg/ml]的 IL -4 水平高于其他组(P<0.01 和 P<0.05)。与 PBS 组和 18K 对照组相比,SWA 刺激后 T - PDDV 组的脾细胞增殖显著增加(P<0.01)。然而,C - T - B 组、PBS 组和 18K 对照组之间的脾细胞增殖无显著差异(P>0.05)。这些发现表明,T - PDDV 和 C - T - B PDDV 混合物诱导的免疫反应比 C - PDDV 或 B - PDDV 更强。