Kalnin Kirill, Chivukula Sudha, Tibbitts Timothy, Yan Yanhua, Stegalkina Svetlana, Shen Lihua, Cieszynski Jacqueline, Costa Victor, Sabharwal Robert, Anderson Stephen F, Christensen Neil, Jagu Subhashini, Roden Richard B S, Kleanthous Harry
Research, Sanofi Pasteur, 38 Sidney Street, Cambridge, MA, USA.
Research, Sanofi Pasteur, Shantha, Hyderabad, AP, India.
Vaccine. 2017 Sep 5;35(37):4942-4951. doi: 10.1016/j.vaccine.2017.07.086. Epub 2017 Aug 1.
To achieve durable and broad protection against human papillomaviruses by vaccination with multimers of minor capsid antigen L2 using self-adjuvanting fusions with the toll-like receptor-5 (TLR5) ligand bacterial flagellin (Fla) instead of co-formulation with alum.
Fla fusions with L2 protective epitopes comprising residues 11-200, 11-88 and/or 17-38 of a single or multiple HPV types were produced in E. coli and their capacity to activate TLR5 signaling was assessed. Immunogenicity was evaluated serially following administration of 3 intramuscular doses of Fla-L2 multimer without exogenous adjuvant, followed by challenge 1, 3, 6 or 12months later, and efficacy compared to vaccination with human doses of L1 VLP vaccines (Gardasil and Cervarix) or L2 multimer formulated in alum. Serum antibody responses were assessed by peptide ELISA, in vitro neutralization assays and passive transfer to naïve rabbits in which End-Point Protection Titers (EPPT) were determined using serial dilutions of pooled immune sera collected 1, 3, 6 or 12months after completing active immunization. Efficacy was assessed by determining wart volume following concurrent challenge at different sites with HPV6/16/18/31/45/58 'quasivirions' containing cottontail rabbit papillomavirus (CRPV) genomes.
Vaccination in the absence of exogenous adjuvant with Fla-HPV16 L2 11-200 fusion protein elicited durable protection against HPV16, but limited cross-protection against other HPV types. Peptide mapping data suggested the importance of the 17-38 aa region in conferring immunity. Indeed, addition of L2 residues 17-38 of HPV6/18/31/39/52 to a Fla-HPV16 L2 11-200 or 11-88 elicited broader protection via active or passive immunization, similar to that seen with vaccination with an alum-adjuvanted L2 multimer comprising the aa 11-88 peptides of five or eight genital HPV types.
Vaccination with flagellin fused L2 multimers provided lasting (>1year) immunity without the need for an exogenous adjuvant. Inclusion of the L2 amino acid 17-38 region in such multi-HPV type fusions expanded the spectrum of protection.
通过使用与Toll样受体5（TLR5）配体细菌鞭毛蛋白（Fla）的自佐剂融合物而非与明矾共同配制，用次要衣壳抗原L2的多聚体进行疫苗接种，以实现对人乳头瘤病毒的持久和广泛保护。
在大肠杆菌中制备与包含一种或多种HPV类型的11 - 200、11 - 88和/或17 - 38位残基的L2保护性表位的Fla融合物，并评估其激活TLR5信号传导的能力。在不使用外源性佐剂的情况下，连续3次肌肉注射Fla - L2多聚体后，分别于1、3、6或12个月后进行攻击，评估免疫原性，并与接种人剂量的L1病毒样颗粒疫苗（加德西和希瑞适）或在明矾中配制的L2多聚体疫苗的效果进行比较。通过肽ELISA、体外中和试验以及将血清被动转移至未免疫的兔子中评估血清抗体反应，在完成主动免疫后1、3、6或12个月收集混合免疫血清，通过系列稀释测定终点保护滴度（EPPT）。通过用含有棉尾兔乳头瘤病毒（CRPV）基因组的HPV6/16/18/31/45/58“准病毒体”在不同部位同时攻击后测定疣体积来评估疫苗效力。
在不使用外源性佐剂的情况下，用Fla - HPV16 L2 11 - 200融合蛋白进行疫苗接种可引发对HPV16的持久保护，但对其他HPV类型的交叉保护有限。肽图谱数据表明17 - 38氨基酸区域在赋予免疫力方面的重要性。实际上，将HPV6/18/31/39/52的L2残基17 - 38添加到Fla - HPV16 L2 11 - 200或11 - 88中，通过主动或被动免疫可引发更广泛的保护，类似于用包含五种或八种生殖器HPV类型的11 - 88氨基酸肽的明矾佐剂L2多聚体进行疫苗接种所观察到的情况。
用鞭毛蛋白融合的L2多聚体进行疫苗接种无需外源性佐剂即可提供持久（>1年）免疫力。在这种多HPV类型融合物中包含L2氨基酸17 - 38区域可扩大保护范围。
