A Dual-Type L2 11-88 Peptide from HPV Types 16/18 Formulated in Montanide ISA 720 Induced Strong and Balanced Th1/Th2 Immune Responses, Associated with High Titers of Broad Spectrum Cross-Reactive Antibodies in Vaccinated Mice.

Farhad Motavalli Khiavi,Majid Golkar,Farzin Roohvand,Arash Arashkia,Kayhan Azadmanesh,Maryam Nasimi

doi:10.1155/2018/9464186

Farhad Motavalli Khiavi, Majid Golkar + Show 4 more

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https://doi.org/10.1155/2018/9464186

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Abstract

E. coli-derived concatenated, multitype L2-conserved epitopes of human papillomavirus (HPV) L2 protein might represent a less expensive and pan-type vaccine alternative (compared to type-specific HPV L1 virus-like particles), if stable protein expression and strong immunogenicity features could be met. Herein, three dual-type- (DT-) HPV L2 fusion peptides comprising the three head-to-tail tandem repeats (multimers) of either HPV 16 epitope “17-36” or “69-81” or one copy (monomer) of 11-88 fused to the same residues of HPV 18 were constructed and expressed in E. coli. SDS-PAGE and Western blot analyses indicated the proper expression and stability of the E. coli-derived DT peptides. Mice immunized by formulation of the purified DT peptides and Freund's adjuvant (CFA/IFA) raised neutralizing antibodies (NAbs; the highest for DT: 11-88 peptide) which showed proper cross-reactivity to HPV types: 18, 16, 31, and 45 and efficiently neutralized HPV 18/16 pseudoviruses in vitro. Immunization studies in mice by formulation of the DT: 11-88 × 1 peptide with various adjuvants (alum, MF59, and Montanides ISA 720 and 50) indicated that Montanide adjuvants elicited the highest cross-reactive titers of NAbs and similar levels of IgG1 and IgG2a (switching towards balanced Th1/Th2 responses). The results implied development of low-cost E. coli-derived DT: 11-88 peptide formulated in human compatible ISA 720 adjuvant as a HPV vaccine.

Highlights

Human papillomavirus (HPV), a heterogeneous group of nonenveloped double-stranded DNA tumor viruses with around 8 kb genome encoding for early and late (L1 and L2 capsids) proteins, infects the epithelia of the skin and mucosa in humans
Restriction analyses of the three constructs harboring the genes corresponding to the dual-type fusion L2 peptide by NcoI/XhoI restriction enzymes followed by agarose gel electrophoresis resulted in two bands of 5350 bp corresponding to the body of digested pET-28a vector and 385 bp, 300 bp, or 500 bp for the fragment inserted into pET-17, pET-69, and pET-88, respectively
Digestion and sequence analyses of all four pET28-HPV L2 (11-200) vectors corresponding to types 16, 18, 31, and 45 resulted in two fragments of 5360 bp corresponding to the body of digested pET-28 vector and 600 bp corresponding to the fragment inserted into pETHPV 16, pET-HPV 18, pET-HPV 31, and pET-HPV 45 (Supplementary Fig. 1)

Summary

Introduction

Human papillomavirus (HPV), a heterogeneous group of nonenveloped double-stranded DNA tumor viruses with around 8 kb genome encoding for early and late (L1 and L2 capsids) proteins, infects the epithelia of the skin and mucosa in humans. More than 200 genotypes of HPV have been identified, among which at least 15 types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, and 82) are responsible for 5% of global cancer cases (especially cervical cancer) [3,4,5]. Persistent infection with four HPV types—16, 18, 31, and 45—is suggested to be the cause of more than 90% of global cervical carcinomas [6, 7]. Worldwide meta-analyses have indicated that two-thirds of women with cervical cancer were infected by either HPV 16 (51%) or HPV 18 (16.2%) genotypes [8]. A study on HPV prevalence and genotypes in various histological subtypes of cervical adenocarcinoma indicated that HPV 16 was the most common of Journal of Immunology Research

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