Expression of HPV6b L1/EBV LMP2 multiepitope and immunogenicity in mice  

作　　者：Shanli Zhu Xiangyang Xue Jianxiao Liu Lijun Lu Pengyun Zhao Jia Wang WenShu Li Lifang Zhang 

机构地区：[1]Department of Microbiology and Immunology, Wenzhou Medical College, Wenzhou 325000, China

出　　处：《Acta Biochimica et Biophysica Sinica》2010年第8期515-521,共7页生物化学与生物物理学报（英文版）

基　　金：This work was supported by a grant from the National Natural Science Foundation of China （30671882）.

摘　　要：Human papillomavirus （HPV） major capsid protein L1 is an important vehicle for the delivery of epitopes. To investigate the expression and immunogenicity of hybridized HPV6b L1 containing multiepitope of EpsteinBarr virus （EBV） latency membrane protein 2 （LMP2）, a recombinant plasmid pcDNA3.1（＋） containing mammalian codon-optimization HPV6b L1 gene and EBV LMP2 multiepitope was constructed. The EBV LMP2 multiepitope containing T- and B-cell epitope-rich peptides was inserted into C-terminal of HPV6b Ll-coding sequence. The constructed plasmid after verified by enzyme restriction assay and DNA sequencing was transfected into COS-7 cells. Expression of the chimeric gene in COS-7 cells was confirmed by RT-PCR, western blot analysis and immunofluorescence assay. Results revealed successful expression of the chimeric HPV6b L1/EBV LMP2 multiepitope gene both at the mRNA and protein levels in transfected COS-7 cells. Intramuscular administration in mice was able to elicit not only antibodies against HPV6b L1 virus-like particle and EBV LMP2, but also cytotoxic T lymphocyte activity against the EBV LMP2 epitopes. The present results confirmed that HPV L1 protein is potential to deliver multiepitope of EBV LMP2 as immunogen to the MHC class I and class II pathways, extend- ing the use of HPV L1 as delivery vehicles.Human papillomavirus （HPV） major capsid protein L1 is an important vehicle for the delivery of epitopes. To investigate the expression and immunogenicity of hybridized HPV6b L1 containing multiepitope of EpsteinBarr virus （EBV） latency membrane protein 2 （LMP2）, a recombinant plasmid pcDNA3.1（＋） containing mammalian codon-optimization HPV6b L1 gene and EBV LMP2 multiepitope was constructed. The EBV LMP2 multiepitope containing T- and B-cell epitope-rich peptides was inserted into C-terminal of HPV6b Ll-coding sequence. The constructed plasmid after verified by enzyme restriction assay and DNA sequencing was transfected into COS-7 cells. Expression of the chimeric gene in COS-7 cells was confirmed by RT-PCR, western blot analysis and immunofluorescence assay. Results revealed successful expression of the chimeric HPV6b L1/EBV LMP2 multiepitope gene both at the mRNA and protein levels in transfected COS-7 cells. Intramuscular administration in mice was able to elicit not only antibodies against HPV6b L1 virus-like particle and EBV LMP2, but also cytotoxic T lymphocyte activity against the EBV LMP2 epitopes. The present results confirmed that HPV L1 protein is potential to deliver multiepitope of EBV LMP2 as immunogen to the MHC class I and class II pathways, extend- ing the use of HPV L1 as delivery vehicles.

关 键 词：human papillomavirus Epstein-Barr virus latency membrane protein EPITOPE 

分 类 号：Q556.3[生物学—生物化学] S858.315.3[农业科学—临床兽医学]