Abstract:Objective To analyze whether different intracellular location of expressed proteins may affect humoral immunity response upon DNA immunization.
Methods EGFP-HPV16 L1 and EGFP-HPV16L1△NLS fusion genes were amplified from pFB-EGFP-HPV16L1 and pFB-EGFP-HPV16L1△NLS plasmid, respectively, then were recombined into pcDNA3.1(+) vector to generate recombinant pcDNA-EGFP-HPV16L1 and pcDNA -EGFP-HPV16L1△NLS plasmid, respectively. The recombinant plasmids were transfected into CHO cells and fluorescences of EGFP tagged fusion proteins were observed under fluorescent microscope. The recombinant plasmids were subsequently injected into the tibial muscles to immunize BALB/c mices and titers of the antisera were determined using ELISA.
Results The EGFP-HPV16 L1 and EGFP-HPV16L1△NLS were correctly inserted into pcDNA3.1 (+) vectors. EGFP-HPV16 L1 fusion protein could be detected in the nucleus of CHO cells, while the EGFP-HPV16L1△NLS fusion protein was located in the cytoplasm of CHO cells. The two recombinant plasmids could induce humoral immunity response in BALB/c mices. After three times of immunization, the A450 values of antisera of pcDNA -EGFP-HPV16L1△NLS plasmid immunized mices group were higher than that of recombinant pcDNA-EGFP-HPV16L1immunized mices group (P < 0.001).
Conclusion Intracellular localization of the proteins expressed by DNA vaccines could affect humoral immunity response upon DNA immunization. Proteins localized in the cytoplasm could induce stronger humoral immunity response compared with proteins localized in the nucleus upon DNA immunization. The conclusion is very useful to design DNA vaccine to induce stronger humoral immunity response or to produce antibody upon DNA immunization.
杨军, 陈晓黎, 王一理, 司履生. 蛋白表达的亚细胞定位影响基因免疫诱导的体液免疫应答水平的研究[J]. 《中国医药生物技术》杂志, 2012, 7(4): 270-275.
YANG Jun, CHEN Xiao-Li, WANG Yi-Li, SI 履Sheng. Effects of intracellular localization of proteins expressed by DNA vaccines on humoral immunity response upon DNA immunization. , 2012, 7(4): 270-275.