Han Kaikai, Xu Lixin, Yan Ruofeng, Song Xiaokai, Li Xiangrui
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China.
Vet Immunol Immunopathol. 2012 Oct 15;149(3-4):177-85. doi: 10.1016/j.vetimm.2012.06.016. Epub 2012 Jun 19.
Owing to its critical functions in worm physiology, glyceraldehyde-3-phosphate dehydrogenase in Haemonchus contortus (HcGAPDH) is potential candidates for vaccine to control haemonchosis. In this study, DNA vaccine expressing HcGAPDH antigen was tested for protection against experimental H. contortus infections in goats. Fifteen goats (9-10 months of age) were allocated into three trial groups. Group 1 was vaccinated with HcGAPDH DNA vaccine twice on days 0 and 14, and then challenged with 5000 infective H. contortus L3 (third stage larvae) on days 28. Group 2 was an unvaccinated positive control group challenged with H. contortus L3 on days 28. Group 3 was an unvaccinated negative control group that was not challenged with L3. By the method of RT-PCR and Western-blot, transcription and expression of HcGAPDH DNA vaccine were identified at local injection sites post immunizations, respectively. After immunization with the DNA vaccine, significantly high levels of serum IgG, serum IgA, mucosal IgA, CD4(+) T lymphocytes and B lymphocytes were generated. Also, increased numbers of blood eosinophils and decreased haemoglobin level after challenge were observed in the vaccinated group. Meanwhile, cumulative mean faecal worm egg counts and worm burdens in vaccinated group were reduced by 34.9% and 37.73%, respectively. In brief, recombinant HcGAPDH DNA vaccine induced partial immune response against H. contortus infection in goats.
由于磷酸甘油醛脱氢酶在捻转血矛线虫生理过程中发挥关键作用,捻转血矛线虫磷酸甘油醛脱氢酶(HcGAPDH)是控制血矛线虫病疫苗的潜在候选对象。在本研究中,对表达HcGAPDH抗原的DNA疫苗预防山羊实验性捻转血矛线虫感染的效果进行了检测。15只山羊(9 - 10月龄)被分为三个试验组。第1组在第0天和第14天用HcGAPDH DNA疫苗免疫两次,然后在第28天用5000条感染性捻转血矛线虫L3(第三期幼虫)进行攻虫。第2组是未免疫的阳性对照组,在第28天用捻转血矛线虫L3进行攻虫。第3组是未免疫的阴性对照组,未用L3进行攻虫。通过RT-PCR和Western-blot方法,分别在免疫后于局部注射部位鉴定了HcGAPDH DNA疫苗的转录和表达情况。用该DNA疫苗免疫后,产生了显著高水平的血清IgG、血清IgA、黏膜IgA、CD4(+) T淋巴细胞和B淋巴细胞。此外,在攻虫后,接种疫苗组观察到血嗜酸性粒细胞数量增加和血红蛋白水平降低。同时,接种疫苗组的粪便虫卵累计平均计数和虫体负荷分别降低了34.9%和37.73%。简而言之,重组HcGAPDH DNA疫苗诱导了山羊对捻转血矛线虫感染的部分免疫反应。