title: Expression of HPV6b L1/EBV LMP2 multiepitope and immunogenicity in mice.
abstract: 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 Epstein-Barr 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 L1-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, extending the use of HPV L1 as delivery vehicles.
authors: Department of Microbiology and Immunology, Wenzhou Medical College, China.
authors: Shanli
authors: S
authors: Zhu
authors: Xiangyang
authors: X
authors: Xue
authors: Jianxiao
authors: J
authors: Liu
authors: Lijun
authors: L
authors: Lu
authors: Pengyun
authors: P
authors: Zhao
authors: Jia
authors: J
authors: Wang
authors: WenShu
authors: W
authors: Li
authors: Lifang
authors: L
authors: Zhang
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D015153:Blotting, Western; D019556:COS Cells; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002470:Cell Survival / Q000276:immunology; D002522:Chlorocebus aethiops; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D052163:Human papillomavirus 6 / Q000235:genetics / Q000378:metabolism; D007114:Immunization / Q000379:methods; D007273:Injections, Intramuscular; D051379:Mice; D008807:Mice, Inbred BALB C; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D000939:Epitopes; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; C114580:HPV L1 protein, Human papillomavirus; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins