目的:通过原位杂交检测技术探索高危型人乳头瘤病毒(HR-HPV)分型检测阴性的子宫颈高级别鳞状上皮内病变(HSIL)是否由人乳头瘤病毒(HPV)感染引起,进而为子宫颈HSIL的精确诊断及治疗提供新思路。方法:收集2020年1月至2022年10月就诊于山...目的:通过原位杂交检测技术探索高危型人乳头瘤病毒(HR-HPV)分型检测阴性的子宫颈高级别鳞状上皮内病变(HSIL)是否由人乳头瘤病毒(HPV)感染引起,进而为子宫颈HSIL的精确诊断及治疗提供新思路。方法:收集2020年1月至2022年10月就诊于山东第二医科大学附属医院妇科行子宫颈活检或手术治疗的305例子宫颈HSIL患者的病历资料,其中HR-HPV分型检测阴性患者为研究组(15例),阳性患者为对照组(290例)。采用RNAscope原位杂交技术检测两组患者样本中HPV E6/E7 m RNA的表达,并使用染色强度半定量评分分析HPV E6/E7 m RNA的表达情况,同时采用免疫组织化学(IHC)方法检测p16、Ki-67的表达情况。结果:(1)305例HSIL患者中,15例(4.9%)HR-HPV分型检测为阴性。(2)研究组中73.3%(11/15)HPV E6/E7 m RNA呈阳性表达,与对照组100.0%(290/290)比较,差异有统计学意义(P<0.001)。(3)研究组p16的阳性表达率与对照组比较,差异有统计学意义(93.3%vs.100.0%,P=0.049),两组Ki-67阳性表达率均为100.0%。(4)组织病理学类型为子宫颈上皮内瘤变(CIN)2的样本中,研究组HPV E6/E7 m RNA总体评分明显低于对照组,差异有统计学意义(P=0.004);CIN3的样本中,研究组HPV E6/E7 m RNA总体评分虽低于对照组,但差异无统计学意义(P=0.158)。结论:子宫颈癌筛查中,单纯采用HR-HPV分型检测可能导致4.9%的HSIL患者因结果阴性而出现漏诊或误诊。HPV E6/E7 m RNA原位杂交检测技术可能有助于降低HR-HPV分型检测在HSIL中的假阴性率,并辅助预测子宫颈病变的发展方向,进而为实现个体化精准治疗提供新的思路。展开更多
Objective:Cervical cancer caused by persistent high-risk human papillomavirus(hrHPV)infection remains a leading cause of cancer-related mortality in women.As prophylactic HPV vaccines cannot eliminate existing infecti...Objective:Cervical cancer caused by persistent high-risk human papillomavirus(hrHPV)infection remains a leading cause of cancer-related mortality in women.As prophylactic HPV vaccines cannot eliminate existing infections,developing therapeutic vaccines targeting HPV E6/E7 oncoproteins is critical for reversing precancerous lesions.This study aimed to design a novel multiepitope vaccine against HPV16,incorporating newly identified immunodominant epitopes and evaluating the therapeutic efficacy.Methods:The multi-epitope vaccine HSP70-12P was bioinformatically designed to include cytotoxic T lymphocyte(CTL)and helper T lymphocyte(HTL)epitopes from HPV16 E6/E7,which were fused to the C-terminal domain(residues 359±610)of Mycobacterium tuberculosis HSP70 as an adjuvant.Two formulations were used,as follows:(1)protein-based Pro-HSP70-12P;and(2)DNA-based DNA-HSP70-12P.Therapeutic efficacy was evaluated in TC-1 tumor-bearing mouse models.Tumor regression,survival rates,and immune correlates(T cell responses and cytokine profiles)were assessed.Immunodominant epitopes were identified using ELISpot.Results:The Pro-HSP70-12P protein vaccine induced strong immune responses and provided lasting antitumor protection.The vaccine activated cell-mediated immunity and stimulated effector memory T cells in the HPV-16-related tumor mouse model,resulting in strong tumor clearance effects.Pro-HSP70-12P demonstrated superior performance compared to the DNA-HSP70-12P vaccine,achieving complete regression of small tumors(diameter<2 mm)with a single dose and conferring long-lasting protection in TC-1 rechallenge experiments.Three novel immunodominant epitopes were identified(E6-38-45,E6-124-132,and E7-50-57).The E6 epitopes address a critical gap in E6-targeted vaccine design.Conclusions:The multi-epitope protein vaccine,Pro-HSP70-12P,represents a potent therapeutic candidate against HPV-driven malignancies,which has the capacity to induce tumor regression and long-term immunity.These findings support further clinical development.展开更多
文摘目的:通过原位杂交检测技术探索高危型人乳头瘤病毒(HR-HPV)分型检测阴性的子宫颈高级别鳞状上皮内病变(HSIL)是否由人乳头瘤病毒(HPV)感染引起,进而为子宫颈HSIL的精确诊断及治疗提供新思路。方法:收集2020年1月至2022年10月就诊于山东第二医科大学附属医院妇科行子宫颈活检或手术治疗的305例子宫颈HSIL患者的病历资料,其中HR-HPV分型检测阴性患者为研究组(15例),阳性患者为对照组(290例)。采用RNAscope原位杂交技术检测两组患者样本中HPV E6/E7 m RNA的表达,并使用染色强度半定量评分分析HPV E6/E7 m RNA的表达情况,同时采用免疫组织化学(IHC)方法检测p16、Ki-67的表达情况。结果:(1)305例HSIL患者中,15例(4.9%)HR-HPV分型检测为阴性。(2)研究组中73.3%(11/15)HPV E6/E7 m RNA呈阳性表达,与对照组100.0%(290/290)比较,差异有统计学意义(P<0.001)。(3)研究组p16的阳性表达率与对照组比较,差异有统计学意义(93.3%vs.100.0%,P=0.049),两组Ki-67阳性表达率均为100.0%。(4)组织病理学类型为子宫颈上皮内瘤变(CIN)2的样本中,研究组HPV E6/E7 m RNA总体评分明显低于对照组,差异有统计学意义(P=0.004);CIN3的样本中,研究组HPV E6/E7 m RNA总体评分虽低于对照组,但差异无统计学意义(P=0.158)。结论:子宫颈癌筛查中,单纯采用HR-HPV分型检测可能导致4.9%的HSIL患者因结果阴性而出现漏诊或误诊。HPV E6/E7 m RNA原位杂交检测技术可能有助于降低HR-HPV分型检测在HSIL中的假阴性率,并辅助预测子宫颈病变的发展方向,进而为实现个体化精准治疗提供新的思路。
基金supported by Chinese Academy Medical Sciences Innovation Fund for Medical Sciences(Grant No.2021-I2M-1-004)The Yunnan Provincial Science and Technology Department(Grant No.202002AA100009)+3 种基金The Yunnan Province Xingdian Talent Support Program(Grant No.XDYC-CYCX-2023-0074)The Yunnan Provincial Innovation Team of Therapeutic Neutralizing Antibody(Grant No.202405AS350026)Yunnan Fundamental Research Projects(Grant No.202201AS070059)NMPA Key Laboratory for Quality Control and Evaluation of Vaccines and Biological Products。
文摘Objective:Cervical cancer caused by persistent high-risk human papillomavirus(hrHPV)infection remains a leading cause of cancer-related mortality in women.As prophylactic HPV vaccines cannot eliminate existing infections,developing therapeutic vaccines targeting HPV E6/E7 oncoproteins is critical for reversing precancerous lesions.This study aimed to design a novel multiepitope vaccine against HPV16,incorporating newly identified immunodominant epitopes and evaluating the therapeutic efficacy.Methods:The multi-epitope vaccine HSP70-12P was bioinformatically designed to include cytotoxic T lymphocyte(CTL)and helper T lymphocyte(HTL)epitopes from HPV16 E6/E7,which were fused to the C-terminal domain(residues 359±610)of Mycobacterium tuberculosis HSP70 as an adjuvant.Two formulations were used,as follows:(1)protein-based Pro-HSP70-12P;and(2)DNA-based DNA-HSP70-12P.Therapeutic efficacy was evaluated in TC-1 tumor-bearing mouse models.Tumor regression,survival rates,and immune correlates(T cell responses and cytokine profiles)were assessed.Immunodominant epitopes were identified using ELISpot.Results:The Pro-HSP70-12P protein vaccine induced strong immune responses and provided lasting antitumor protection.The vaccine activated cell-mediated immunity and stimulated effector memory T cells in the HPV-16-related tumor mouse model,resulting in strong tumor clearance effects.Pro-HSP70-12P demonstrated superior performance compared to the DNA-HSP70-12P vaccine,achieving complete regression of small tumors(diameter<2 mm)with a single dose and conferring long-lasting protection in TC-1 rechallenge experiments.Three novel immunodominant epitopes were identified(E6-38-45,E6-124-132,and E7-50-57).The E6 epitopes address a critical gap in E6-targeted vaccine design.Conclusions:The multi-epitope protein vaccine,Pro-HSP70-12P,represents a potent therapeutic candidate against HPV-driven malignancies,which has the capacity to induce tumor regression and long-term immunity.These findings support further clinical development.
