Development of a Pan-Oncogenic HPV Preventive Vaccine

两种HPV衣壳蛋白L1和L2是不依赖于肉毒杆菌素的保护性抗原。HPV L1病毒样颗粒（VLP）疫苗接种可诱导中和抗体，并在强型限制患者中产生保护作用。对>15种已知致癌HPV的广泛保护对于最终停止细胞学筛查和根除宫颈癌是必要的。我们建议L2作为一个单一的保守的保护 抗原的我们已经在兔和小鼠中显示，用细菌中产生的L2 11-200接种疫苗保护抵抗同源病毒类型以及进化上远距离的异源类型（即，用HPV 16 L2 11-200接种疫苗显著地保护抵抗兔乳头瘤病毒感染），支持基于L2的泛致癌HPV疫苗的可能性。此外，我们已经表明，接种L2诱导 患者体内的交叉中和抗体与目前的多价L1 VLP疫苗不同，在E.大肠杆菌的生产成本低廉。因此，生产成本较低的泛致癌HPV类型将在美国和发展中国家服务不足的地区产生最大的影响。快速获得预防干预发展（RAPID，NCI）计划正在生产GMP级 包含串联的HPV 6、16和18的L2 11-200（L2 11- 200 × 3）与明矾佐剂的融合蛋白用于该提出的临床试验。假设1：用明矾佐剂中的HPV L2 11- 200 x3多肽对患者进行疫苗接种是安全的。具体目标1：进行一项双盲、安慰剂对照、剂量递增的I期试验，以评估健康女性中HPV L2 11- 200 x3多肽疫苗接种的安全性。假设2： 用明矾佐剂中的HPV L2 11- 200 x3多肽对患者进行疫苗接种诱导高滴度的广泛中和抗体。具体目标#2：确定明矾中HPV L2 11- 200 x3的剂量范围，与Gardasil相比，中和抗体的滴度以及中和的HPV类型谱。HPV感染仅限于上皮细胞，不会进入血流，但中和IgG被动转移到血流中具有保护作用。抗体在哪里相遇并中和 人乳头瘤病毒？假设3：L2特异性HPV中和抗体渗出到生殖道是保护的相关因素。具体目标#3：为了确定从接种HPV L2 11- 200 x3、HPV 16 L1衣壳或Gardasil的患者被动转移具有IgG或IgM的小鼠是否会保护小鼠免受HPV假病毒体的阴道攻击，并比较保护所需的最小中和抗体滴度。

The two HPV capsid proteins, L1 and L2, are botin independent protective antigens. Vaccination with) HPV L1 virus-like particles (VLP) Induces neutralizing antibodies and protection in patients with strong type restriction. Broad protection against the >15 known oncogenic HPVs is necessary for the eventual cessation of cytologic screening and eradication of cervical cancer. We propose L2 as a single conserved protective antigen. We have shown in rabbits and mice that vaccination with L2 11-200 produced in bacteria protects against both the homologous virus type as well as evolutionarily distant heterologous types (i.e. remarkably vaccination with HPVl 6 L2 11-200 protects against rabbit papillomavirus infection) supporting the possibility of an L2-based pan-oncogenic HPV vaccine. Furthermore, we have shown that vaccination with L2 induces cross-neutralizing antibodies in patients. Unlike current multivalent LI VLP vaccines, a single L2-based antigen produced in E. coli is inexpensive to produce. As such, a pan-oncogenic HPV type that is less costly to produce would have its greatest impact in underserved areas in the US and in developing nations. The Rapid Access to Preventive Intervention Development (RAPID, NCI) program is producing GMP-grade fusion protein comprising L2 11-200 of HPV6, 16 and 18 in tandem (L2 11-200x3) with alum adjuvant for this proposed clinical trial. HYPOTHESIS 1: Vaccination of patients with HPV L2 11-200x3 polypeptide in alum adjuvant is safe. Specific Aim #1: To perform a double-blinded, placebo-controlled, dose escalating Phase I trial to evaluate the safety HPV L2 11-200x3 polypeptide vaccination in healthy women. HYPOTHESIS 2: Vaccination of patients with HPV L2 11-200x3 polvpeptide in alum adjuvant induces high titers of broadly neutralizing antibodies. Specific Aim #2: To determine the dose ranging for HPV L2 11-200x3 in alum with respect to titers of neutralizing antibody and also the spectrum of HPV types neutralized in comparison to Gardasil¿. HPV infection is restricted to the epithelium and does not enter the bloodstream, yet passive transfer of neutralizing IgG into the bloodstream is protective. Where does the antibody meet and neutralize HPV? HYPOTHESIS 3: Transudation of L2-specific HPV neutralizing antibody into the genital tract is the relevant correlate of protection. Specific Aim #3: To determine whether passive transfer of mice with IgG or IgM from patients vaccinated with HPV L2 11-200x3, HPV16 LI capsomeres or Gardasil will confer protection from vaginal challenge of mice with HPV pseudovirion and compare the minimal neutralizing antibody titers required for protection.