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p16 Immunohistochemistry is useful in confirming high-grade squamous intraepithelial lesions (HSIL) in women with negative HPV testing.

SUMMARY: It is believed that almost all squamous cell carcinomas of the cervix are associated with HR-HPV infection. However, a subset of high-grade squamous intraepithelial lesion (HSIL) (CIN2 and CIN3) lesions is found in those women with negative HPV testing. Knowledge of HPV status can influence pathologists' decision in rendering the diagnosis of cervical squamous intraepithelial lesions (SIL). p16, a surrogate marker for HSIL, has been widely applied to facilitate accurate diagnosis of HPV-related cervical dysplasia, especially CIN2 and CIN3. To assess whether p16 immunostaining is useful in diagnosing HSIL in women with negative HPV testing, we studied the utility of p16 immunohistochemistry in 46 women of HSIL and HPV-negative status. A total of 46 cases of initial biopsies with histopathologically diagnosed HSIL (CIN2 and CIN3) were identified from our hospital archives. All women were HPV negative with at least 1 HPV testing using HC-II (Qiagen) within 6 mo of initial biopsy. LEEP procedures within 6 mo of initial biopsies were reviewed and documented. Immunohistochemical staining of p16 was performed on recuts of all original biopsies. Some LEEP specimens without evidence of HSIL (CIN2 and CIN3) on hematoxylin and eosin had recuts with deeper levels and p16 immunostaining to confirm the negative diagnosis. p16 immunostaining were evaluated as negative, focal/patchy, or diffuse staining pattern. Patients' HPV testing status and related clinicopathologic information were reviewed, tabulated, and correlated with p16 immunostaining patterns. Forty-six women between the age of 17 and 58 yr, with a median of 35 yr, were all HPV-negative. All women, except 2, had an abnormal cytologic interpretation at the time of HPV testing ranging from ASC-US to HSIL. Forty-two women (91.3%) had LEEP procedures done within 6 mo of the initial biopsies. LEEP specimens showed that 76.2% (32 cases) women had HSIL, including 22 cases of CIN2 and 10 cases of CIN3, 14.3% (6 cases) had low-grade squamous intraepithelial lesion (CIN1), and 9.5% (4 cases) had benign cervix. p16 immunostaining, performed on initial biopsies with histopathologic diagnoses of CIN2 or CIN3, showed that 66.7% (28 cases) had diffuse staining pattern, 16.7% (7 cases) had focal/patchy pattern, and 16.7% (7 cases) had negative p16 staining. On LEEP follow-up, all 28 cases with diffuse p16 staining pattern had HSIL (CIN2 and CIN3), and all 7 cases with negative p16 staining had no detectable high-grade dysplasia. For those 7 cases with focal/patch p16 staining pattern, 4 had HSIL (CIN2) and 3 had low-grade squamous intraepithelial lesion (CIN1) on LEEP follow-up. Approximately 76% of women with negative HPV and diagnosis of HSIL (CIN2 and CIN3) on initial biopsy had confirmed HSIL (CIN2 and CIN3) in subsequent LEEP follow-up. Diffuse p16 immunostaining pattern is the hallmark of HSIL because it correlates 100% with CIN2 and CIN3 lesions between initial biopsy and LEEP specimens, regardless of the HPV status. The negative predictive value for p16 immunoreactivity to predict cervical lesions less than high grade is almost 100% in our study. Our study suggests that when a woman is negative for HPV and also negative for p16, diagnosis of HSIL should be very cautious in void of unnecessary LEEP procedures.

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