Yang Heng, Cao Sanjie, Huang Xiaobo, Liu Jiawen, Tang Ying, Wen Xintian
Sichuan Agricultural University, Ya'an, China.
Vaccine. 2009 Aug 13;27(37):5035-40. doi: 10.1016/j.vaccine.2009.06.050. Epub 2009 Jun 30.
Attenuated Salmonella typhimurium was selected as a transgenic vehicle for the development of live mucosal vaccines against transmissible gastroenteritis virus (TGEV). A 2.2kb DNA fragment, encoding for N-terminal domain glycoprotein S of TGEV, was amplified by RT-PCR and cloned into eukaryotic expression vector pVAX1. The recombinant plasmid pVAX-S was transformed by electroporation into attenuated S. typhimurium SL7207, the expression and translation of the pVAX-S delivered by recombinant S. typhimurium SL7207 (pVAX-S) was detected in vitro and in vivo respectively. BALB/c mice were inoculated orally with SL7207 (pVAX-S) at different dosages, the bacterium was safe to mice at dosage of 2x10(9)CFU and eventually eliminated from the spleen and liver at week 4 post-immunization. Mice immunized with different dosages of SL7207 (pVAX-S) elicited specific anti-TGEV local mucosal and humoral responses as measured by indirect ELISA assay. Moreover, the immunogenicity of the DNA vaccine was highly dependent on the dosage of the attenuated bacteria used for oral administration, 10(9)CFU dosage group showed higher antibody response than 10(8)CFU and 10(7)CFU dosages groups during week 4-8 post-immunization. The results indicated that attenuated S. typhimurium could be used as a delivery vector for oral immunization of TGEV DNA vaccine.
减毒鼠伤寒沙门氏菌被选为开发针对传染性胃肠炎病毒(TGEV)的活黏膜疫苗的转基因载体。通过RT-PCR扩增出编码TGEV N端结构域糖蛋白S的2.2kb DNA片段,并将其克隆到真核表达载体pVAX1中。将重组质粒pVAX-S通过电穿孔转化到减毒鼠伤寒沙门氏菌SL7207中,分别在体外和体内检测重组鼠伤寒沙门氏菌SL7207(pVAX-S)递送的pVAX-S的表达和翻译。用不同剂量的SL7207(pVAX-S)口服接种BALB/c小鼠,该菌在2x10(9)CFU剂量下对小鼠安全,并且在免疫后第4周最终从脾脏和肝脏中清除。通过间接ELISA测定法,用不同剂量的SL7207(pVAX-S)免疫的小鼠引发了特异性的抗TGEV局部黏膜和体液反应。此外,DNA疫苗的免疫原性高度依赖于用于口服给药的减毒细菌的剂量,在免疫后第4 - 8周,10(9)CFU剂量组比10(8)CFU和10(7)CFU剂量组显示出更高的抗体反应。结果表明,减毒鼠伤寒沙门氏菌可作为TGEV DNA疫苗口服免疫的递送载体。