评估在口腔冲洗液和 Merocel®海绵中收集的唾液中 HPV-16 和 HPV-18 特异性抗体的测量。
Evaluation of HPV-16 and HPV-18 specific antibody measurements in saliva collected in oral rinses and merocel® sponges.
机构信息
HPV Immunology Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research sponsored by the National Cancer Institute, Frederick, MD, USA.
Center for Infection Research in Cancer, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
出版信息
Vaccine. 2018 May 3;36(19):2705-2711. doi: 10.1016/j.vaccine.2018.03.034. Epub 2018 Apr 6.
BACKGROUND
Current Human papillomavirus (HPV) L1 VLP vaccines protect against HPV-16 and HPV-18-associated cancers, in females and males. Although correlates of protection have not been identified, HPV-specific antibodies at sites of infection are thought to be the main mechanism of protection afforded by vaccination. Oral sampling has gained increased attention as a potential alternative to serum in monitoring immunity to vaccination and understanding local immunity in oral cancers.
METHODS
Serum was collected via venipuncture, and saliva was collected via oral rinses and Merocel® sponges from healthy volunteers: 16 unvaccinated females, 6 females (ages 24-41) and 6 mid-adult aged males (ages 27-45) recipients of three doses of the HPV-16/18/6/11 vaccine (Gardasil®). Mid-adult male vaccine trial participants were compared to female participants. Samples were tested for anti-HPV-16 and anti-HPV-18 immunoglobulin G levels by an L1 virus-like particle-based enzyme-linked immunosorbent assay (ELISA).
RESULTS
All vaccinated participants had detectable serum anti-HPV-16 and anti-HPV-18 antibodies. Optimal standard concentration range and sample serial dilutions for oral rinses were determined. The standard curve was not affected by the type of solution examined. Reproducibility of HPV-16 and HPV-18 antibody titers in mouthwash (overall CV < 10%) or in Merocel® extraction buffer was robust (CV < 13%). Excellent assay linearity (R > 0.9) was observed for sera spiked controls in both solutions. HPV-16 and HPV-18 specific antibodies were detectable in saliva from vaccine recipients, both in mouthwash and in Merocel® sponges but levels were several logs lower than those in serum.
CONCLUSIONS
This study confirms the application of HPV-16 and HPV-18 ELISAs currently used in sero-epidemiological studies of immunogenicity of HPV vaccines for use with oral samples. Oral samples may be a useful resource for the detection of HPV-16 and HPV-18-specific antibodies in saliva following vaccination.
背景
目前的人乳头瘤病毒（HPV）L1 VLP 疫苗可预防女性和男性的 HPV-16 和 HPV-18 相关癌症。尽管尚未确定保护相关因素，但感染部位的 HPV 特异性抗体被认为是疫苗提供保护的主要机制。口腔采样作为监测疫苗免疫和了解口腔癌局部免疫的替代血清方法，已得到越来越多的关注。
方法
通过静脉穿刺采集血清，通过口腔冲洗和 Merocel®海绵采集唾液，健康志愿者包括 16 名未接种疫苗的女性、6 名（年龄 24-41 岁）和 6 名中年男性（年龄 27-45 岁），他们接种了三剂 HPV-16/18/6/11 疫苗（Gardasil®）。将中年男性疫苗试验参与者与女性参与者进行比较。通过基于 L1 病毒样颗粒的酶联免疫吸附试验（ELISA）检测血清中抗 HPV-16 和抗 HPV-18 免疫球蛋白 G 水平。
结果
所有接种疫苗的参与者均检测到血清抗 HPV-16 和抗 HPV-18 抗体。确定了口腔冲洗液的最佳标准浓度范围和样品系列稀释度。标准曲线不受所检查溶液类型的影响。口漱剂（总体 CV<10%）或 Merocel®提取缓冲液中 HPV-16 和 HPV-18 抗体滴度的重现性良好（CV<13%）。两种溶液中添加的血清对照品均具有良好的检测线性（R>0.9）。在疫苗接种者的唾液中可检测到 HPV-16 和 HPV-18 特异性抗体，无论是在口漱剂还是在 Merocel®海绵中，但水平比血清低几个对数级。
结论
本研究证实，目前用于 HPV 疫苗免疫原性血清流行病学研究的 HPV-16 和 HPV-18 ELISA 可用于口腔样本。口腔样本可能是一种有用的资源，可用于检测接种疫苗后唾液中的 HPV-16 和 HPV-18 特异性抗体。
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