Expression of HPV6b L1/EBV LMP2 multiepitope and immunogenicity in mice.
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.
Zhu
Shanli
S
Department of Microbiology and Immunology, Wenzhou Medical College, China.
Xue
Xiangyang
X
Liu
Jianxiao
J
Lu
Lijun
L
Zhao
Pengyun
P
Wang
Jia
J
Li
WenShu
W
Zhang
Lifang
L
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
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