Ewaisha Radwa, Ruffin Mack T, Williams Stacy, Chung Yunro, DeGraffinreid Cecilia R, Paskett Electra D, Reiter Paul L, Qiu Ji, Brenner Dean E, Anderson Karen S
Department of Microbiology and Immunology, Faculty of Pharmacy, Alexandria University, Alexandria, Egypt.
Department of Family and Community Medicine, Penn State College of Medicine, Hershey, PA, United States.
Front Immunol. 2025 Jul 31;16:1612761. doi: 10.3389/fimmu.2025.1612761. eCollection 2025.
Understanding the humoral immune response to HPV is important for understanding the natural history of infection and developing biomarkers for early detection of cervical cancer. This has been technically limited by HPV type diversity and challenges of high-throughput protein expression and display. This study aimed to profile the humoral immune response to the proteomes of 12 HPV types in women with or without abnormal cervical cytology undergoing cervical cancer screening.
To detect serum antibodies (Abs) against HPV, we developed custom HPV high-density diffusion-free nucleic acid programmable protein arrays (HD-NAPPA) displaying the proteomes of 2 low-risk (HPV6 and 11) and 10 high-risk (HR) HPV types (HPV16, 18, 31, 33, 35, 39, 45, 51, 52 and 58). Arrays were probed with sera from women undergoing screening for cervical cancer, with normal (n=82) or abnormal (n=54) cervical cytology. HPV DNA testing and typing were done on cytology samples from all participants using an assay that detects 37 HPV types.
Abs to any HPV protein were detected in 47.6% (95% C.I.: 36.5-58.8%) and 40.7% (95% C.I.: 27.9-54.9%) of women with normal and abnormal cytology, respectively and in 44.9% (95% C.I.: 36.4-53.6%) of all women. HPV16 DNA was the most frequently detected type (36.8%, 95% C.I.: 27.4-47.4%), however, Abs against HPV16 were remarkably the least frequently detected (7.4%, 95% C.I.: 3.8-13.5%). The most frequently detected Abs were against L1, in 30.1% (95% C.I.: 22.7-38.7%) of all women (31.7% and 27.8% of women with normal and abnormal Pap, respectively). Abs against E1 and E4 were the most (in 24.3%, 95% C.I.: 17.5-32.5%) and least (13.2%, 95% C.I.: 8.2-20.4%) frequently detected E-Abs in all women, respectively. Among all subjects with antibodies to either L1 or L2, 39.0% (95% C.I.: 24.6-55.5%) of those with L1 antibodies and 51.9% (95% C.I.: 32.4-70.8%) of those with L2 antibodies were positive for the antigen from only one HPV type.
Our findings shed light on the kinetics of HPV-specific humoral immunity in women with normal or abnormal cervical cytology and highlight the need for comprehensive immune profiling in different health and disease stages.
了解针对人乳头瘤病毒（HPV）的体液免疫反应对于理解感染的自然史以及开发宫颈癌早期检测的生物标志物至关重要。这在技术上受到HPV类型多样性以及高通量蛋白质表达和展示挑战的限制。本研究旨在剖析接受宫颈癌筛查的宫颈细胞学正常或异常女性对12种HPV类型蛋白质组的体液免疫反应。
为检测针对HPV的血清抗体（Abs），我们开发了定制的HPV高密度无扩散核酸可编程蛋白质阵列（HD-NAPPA），展示2种低风险（HPV6和11）和10种高风险（HR）HPV类型（HPV16、18、31、33、35、39、45、51、52和58）的蛋白质组。用接受宫颈癌筛查、宫颈细胞学正常（n = 82）或异常（n = 54）女性的血清检测阵列。使用可检测37种HPV类型的检测方法对所有参与者的细胞学样本进行HPV DNA检测和分型。
宫颈细胞学正常和异常的女性中，分别有47.6%（95%置信区间：36.5 - 58.8%）和40.7%（95%置信区间：27.9 - 54.9%）检测到针对任何HPV蛋白的抗体，所有女性中这一比例为44.9%（95%置信区间：36.4 - 53.6%）。HPV16 DNA是最常检测到的类型（36.8%，95%置信区间：27.4 - 47.4%），然而，针对HPV16的抗体却是最不常检测到的（7.4%，95%置信区间：3.8 - 13.5%）。最常检测到的抗体是针对L1的，在所有女性中占30.1%（95%置信区间：22.7 - 38.7%）（宫颈涂片正常和异常的女性分别占31.7%和27.8%）。针对E1和E4的抗体分别是所有女性中最常（24.3%，95%置信区间：17.5 - 32.5%）和最不常（13.2%，95%置信区间：8.2 - 20.4%）检测到的E抗体。在所有对L1或L2有抗体的受试者中，有L1抗体的受试者中39.0%（95%置信区间：24.6 - 55.5%）以及有L2抗体的受试者中51.9%（95%置信区间：32.4 - 70.8%）仅对一种HPV类型的抗原呈阳性。
我们的研究结果揭示了宫颈细胞学正常或异常女性中HPV特异性体液免疫的动力学，并强调了在不同健康和疾病阶段进行全面免疫剖析的必要性。
