多房棘球绦虫重组BCG—Em14—3—3疫苗对小鼠脾细胞凋亡的影响

Effects of recombinant BCG-Eml4-3-3 vaccine on apoptosis of spleen cells in mice infected with Echinococcus multilocularis

目的 检测多房棘球绦虫（Em）重组BCG—Em14—3—3疫苗对原头蚴攻击小鼠脾细胞凋亡的影响。方法 用重组BCG—Em14—3—3疫苗免疫Balb／c小鼠8周后，用50个多房棘球绦虫原头蚴腹腔注射进行攻击，攻击感染后18周剖杀小鼠．分离脾细胞，用刀豆素A（ConA）刺激培养16～18h，然后用Annexin V—FITC染色法检测脾细胞凋亡。结果 小鼠脾细胞原液组和ConA刺激组细胞凋亡发生率均显著低于磷酸缓冲液（PBS）对照组（P〈0．05或〈0．01）：鼻腔黏膜接种组脾细胞凋亡率显著低于皮下注射组（P〈0．01）；重组BCG疫苗组脾细胞凋亡率显著低于pCD—Em10 DNA疫苗肌肉注射组（P〈0．01）。结论 多房棘球绦虫重组BCG—Em14—3—3疫苗可抑制感染鼠脾细胞发生凋亡．增加CD4^＋细胞亚群的数量，加强宿主抗多房棘球绦虫感染的保护性免疫反应。

Objective To detect the effects of recombinant BCG-Eml4-3-3 vaccine on apoptosis of spleen cells in mice challenged with protoscoleces of Echinococcus multilocularis. Methods rBCG-Em14-3-3 vaccine was used to immunize Balb/c mice. 8 weeks after, all the mice were challenged with 50 protoscoleces of Echinococcns multilocularis. On the 18th week of infection, mice were killed for isolation of spleen cells. Cultured in ConA for 16 - 18 hours, spleen cells were detected for apoptosis using Annexin V-FITC staining methods. Results Apoptosis of spleen cells of groups with or without ConA stimulation decreased obviously compared with PBS control group （P 〈 0.05 or 〈 0.01 ）. It was also lower in nasal mucous inoculation group than in subcutaneous inoculation group and in recombinant BCG vaccine group than in pCD-Eml0 DNA muscular injection group（P 〈 0.01）. Conclusions rBCG-Eml4-3-3 vaccine may inhibit apoptosis of spleen cells in rats infected with Echinococcus multilocularis and increase the number of CD4＋ T cells, enhance the host immunity against the challenge of protoscoleces of Echinacoccas multilocularis.