Davidson Emma J, Boswell Christopher M, Sehr Peter, Pawlita Michael, Tomlinson Anne E, McVey Rhona J, Dobson Jennifer, Roberts John St C, Hickling Julian, Kitchener Henry C, Stern Peter L
Immunology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, United Kingdom.
Cancer Res. 2003 Sep 15;63(18):6032-41.
This study assessed the immunological and clinical responses of women with human papillomavirus (HPV) 16-associated high-grade vulval intraepithelial neoplasia (VIN) vaccinated with TA-HPV, a recombinant vaccinia virus encoding modified HPV 16 and 18 E6 and E7. Eighteen women with HPV 16-positive high-grade VIN were vaccinated with TA-HPV. The extent of their baseline disease was compared after 24 weeks by lesion measurements and histological analysis. Viral load was assessed pre- and postvaccination by real time PCR. Cell-mediated immunity to HPV 16 E6 and/or E7 peptides (HLA-A2 epitopes) or vaccinia-infected cell lysates was determined by IFN-gamma enzyme-linked immunospot (ELISPOT) and T cell proliferation using an HPV 16 L2E6E7 fusion protein. Antibodies were measured by ELISA using vaccinia-infected cell lysates or HPV 16 and 18 E6 and E7 glutathione S-transferase-fusion proteins. Lesion-infiltrating CD4(+), CD8(+), CD1a(+), and CD68(+) immune cells were assessed by immunohistochemistry. The single vaccination with TA-HPV was well tolerated, and all patients showed an increased ELISPOT and/or antibody response to vaccinia. There were significant differences in HPV-16 E7-specific ELISPOT and L2E6E7 proliferative responses in the patients at one or more time points postvaccination as compared with the prevaccination status; two patients showed transient increased antibody responses. Overall, 13 women showed an increased HPV 16-specific immune response by one or more methodologies after immunization. Eight patients demonstrated a reduction in lesion diameter of at least 50% and a further four patients showed significant symptom relief. Viral load was reduced or cleared in six of eight lesion responders but also in six of ten nonresponders. Before vaccination, clinical responders had significantly higher levels of lesion-associated CD4(+), CD8(+), and CD1a(+)-immune cells than nonresponders. There were no differences in CD68 (macrophages) between responders and nonresponders before or after vaccination. Nonresponders did show a significant increase in CD4(+)- and CD8(+)- but not CD1a(+)-immune cells postvaccination but at lower levels overall than responder patients. Local immune infiltration may be a critical factor in potential responsiveness to vaccine therapy in HPV-associated neoplasia and should be carefully monitored in future placebo-controlled trials of immunotherapy for VIN.
本研究评估了接种TA-HPV（一种编码修饰型人乳头瘤病毒16型和18型E6和E7的重组痘苗病毒）的人乳头瘤病毒（HPV）16型相关的高级别外阴上皮内瘤变（VIN）女性的免疫和临床反应。18名HPV 16阳性高级别VIN女性接种了TA-HPV。在24周后，通过病变测量和组织学分析比较她们的基线疾病程度。通过实时PCR评估接种前后的病毒载量。通过干扰素-γ酶联免疫斑点法（ELISPOT）以及使用HPV 16 L2E6E7融合蛋白的T细胞增殖来测定对HPV 16 E6和/或E7肽（HLA-A2表位）或痘苗感染细胞裂解物的细胞介导免疫。使用痘苗感染细胞裂解物或HPV 16和18 E6和E7谷胱甘肽S-转移酶融合蛋白通过酶联免疫吸附测定（ELISA）来测量抗体。通过免疫组织化学评估病变浸润的CD4(+)、CD8(+)、CD1a(+)和CD68(+)免疫细胞。单次接种TA-HPV耐受性良好，所有患者对痘苗的ELISPOT和/或抗体反应均增强。与接种前状态相比，接种后一个或多个时间点患者的HPV-16 E7特异性ELISPOT和L₂E6E₇增殖反应存在显著差异；两名患者显示出短暂的抗体反应增强。总体而言，13名女性在免疫后通过一种或多种方法显示出HPV 16特异性免疫反应增强。8名患者的病变直径减小了至少50%，另外4名患者症状明显缓解。8名病变反应者中有6名病毒载量降低或清除，10名无反应者中也有6名病毒载量降低或清除。接种前，临床反应者病变相关的CD4(+)、CD8(+)和CD1a(+)免疫细胞水平显著高于无反应者。接种前后，反应者和无反应者之间的CD68（巨噬细胞）无差异。无反应者接种后CD4(+)和CD8(+)免疫细胞显著增加，但CD1a(+)免疫细胞未增加，且总体水平低于反应者患者。局部免疫浸润可能是HPV相关肿瘤对疫苗治疗潜在反应性的关键因素，在未来VIN免疫治疗的安慰剂对照试验中应仔细监测。
