Using p16 immunohistochemistry to classify morphologic cervical intraepithelial neoplasia 2: correlation of ambiguous staining patterns with HPV subtypes and clinical outcome

Hum Pathol. 2017 Aug:66:144-151. doi: 10.1016/j.humpath.2017.06.014. Epub 2017 Jul 11.

Abstract

p16INK4a immunohistochemistry (IHC) is widely used to facilitate the diagnosis of human papillomavirus (HPV)-associated cervical precancerous lesions. Although most p16 results are distinctly positive or negative, certain ones are ambiguous: they meet some but not all requirements for the "block-positive" pattern. It is unclear whether ambiguous p16 immunoreactivity indicates oncogenic HPV infection or risk of progression. Herein, we compared HPV genotypes and subsequent high-grade squamous intraepithelial lesion (HSIL) outcomes among 220 cervical biopsies with a differential diagnosis of cervical intraepithelial neoplasia 2 based on hematoxylin and eosin morphology and varying degrees of p16 immunoreactivity. p16 results were classified as block positive (n=40, 18%), negative (n=130, 59%), or ambiguous (n=50, 23%), a category we further grouped into 3 patterns: strong/basal (n=18), strong/focal (n=15), and weak/diffuse (n=17). Seventy percent of ambiguous p16 lesions were negative for the most common low- and high-risk HPV types; the remaining 30% were positive for HPV 16, 18, 45, 58, 59, or 66. Three patterns revealed comparably low HPV detection rates (28%, 27%, and 35%). During 12-month surveillance, HSILs were detected in 35% of the p16 block-positive group, 1.5% of negative group, and 16% of the ambiguous group. The accuracy of ambiguous p16 immunoreactivity in predicting oncogenic HPV and HSIL outcome is significantly lower than that of the block-positive pattern but greater than negative staining. Specific guidelines for this intermediate category should prevent diagnostic errors and help implement p16 IHC in general practice.

Keywords: Ambiguous patterns; CIN 2; HSIL; Human papillomavirus; p16 immunohistochemistry.

MeSH terms

  • Biomarkers, Tumor / analysis*
  • Biopsy
  • Cyclin-Dependent Kinase Inhibitor p16 / analysis*
  • DNA, Viral / genetics
  • Disease Progression
  • Female
  • Human Papillomavirus DNA Tests
  • Humans
  • Immunohistochemistry*
  • Neoplasm Grading
  • Papillomaviridae / classification*
  • Papillomaviridae / genetics
  • Papillomavirus Infections / metabolism*
  • Papillomavirus Infections / pathology
  • Papillomavirus Infections / virology
  • Predictive Value of Tests
  • Reproducibility of Results
  • Retrospective Studies
  • Risk Factors
  • Squamous Intraepithelial Lesions of the Cervix / metabolism*
  • Squamous Intraepithelial Lesions of the Cervix / pathology
  • Squamous Intraepithelial Lesions of the Cervix / virology
  • Time Factors
  • Uterine Cervical Dysplasia / chemistry*
  • Uterine Cervical Dysplasia / pathology
  • Uterine Cervical Dysplasia / virology
  • Uterine Cervical Neoplasms / chemistry*
  • Uterine Cervical Neoplasms / pathology
  • Uterine Cervical Neoplasms / virology

Substances

  • Biomarkers, Tumor
  • CDKN2A protein, human
  • Cyclin-Dependent Kinase Inhibitor p16
  • DNA, Viral