This study systematically evaluated the staining performance and reproducibility of three commonly used p16 antibody clones (E6H4, JC8, 6H12) in gynecologic tumors. All clones demonstrated excellent performance on standardized automated staining platforms, achieving 100% positivity/negativity concordance, providing critical reference data for clinical laboratories.
Literature Overview
This article, 'Comparative Evaluation of Three Primary Antibody Clones for p16 Immunohistochemistry in Gynecologic Tumors' published in Antibodies, reviews and summarizes the staining consistency and reproducibility of three p16 antibody clones in gynecologic tumor immunohistochemistry. Through whole-slide and tissue microarray validation, the study demonstrates highly consistent staining results between clones, offering practical evidence for clinical diagnosis.
Background Knowledge
p16 immunohistochemistry (IHC) serves as a surrogate marker for high-risk human papillomavirus (hrHPV) infection, and is widely used in gynecologic tumor diagnostics. p16 protein is overexpressed during HPV infection due to E7 protein-mediated inactivation of pRb, enabling clear differentiation between HPV-associated and non-HPV-associated tumors. However, differences in staining performance between antibody clones may affect result interpretation. This study evaluates the consistency of E6H4, JC8, and 6H12 clones across three fully automated IHC platforms (Ventana/Roche, Agilent/Dako, Leica) to provide standardized data for clinical laboratories. Current research directions include optimization of molecular markers for gynecologic tumors, investigation of p16 expression mechanisms in HPV-negative tumors, and technical stability between clones. While existing studies have assessed p16 performance in head and neck tumors, data in gynecologic tumors remain limited, which this study addresses.
Research Methods and Experiments
The study retrospectively analyzed 176 gynecologic tumor samples, including 42 whole-slide sections and 134 tissue microarray (TMA) cores. Three p16 antibody clones (E6H4, JC8, 6H12) were tested on vendor-recommended automated staining platforms. All samples underwent standardized IHC processing, with staining consistency and reproducibility evaluated by two independent pathologists through blinded assessments. Staining results were classified as block-type positive, mosaic positive, or negative based on ≥70% tumor cell staining intensity.
Key Conclusions and Perspectives
Research Significance and Prospects
This study provides direct evidence for clone interchangeability in gynecologic tumor p16 testing, supporting cross-laboratory result comparisons under standardized conditions. Future research should further evaluate performance of novel clones across platforms and investigate non-HPV-driven roles of p16 in HPV-negative tumors.
Conclusion
This study demonstrates that the three p16 antibody clones (E6H4, JC8, 6H12) exhibit complete consistency and high reproducibility in automated immunohistochemical staining for gynecologic tumors. The findings provide practical guidance for clinical laboratories regarding antibody substitution, standardized implementation, and cost management. Although non-specific staining occurs in some tissues, it does not compromise primary interpretation criteria. We recommend laboratories perform internal validation and integrate external quality control systems when adopting new clones to ensure result reliability. This study contributes critical data for standardizing p16 testing protocols in gynecologic tumor diagnostics.
