Reevaluation of negative cervical conizations: Frequency, diagnostic errors, risk factors and management.

OBJECTIVE: Cervical conization is an effective treatment for precancerous lesions. However, in cases where no high-grade lesion is identified in the surgical specimen, managing these patients may be challenging due to the absence of established follow-up protocols for negative conizations. This study aimed to assess the negative conization rates at our institution by histopathological review, identify diagnostic errors, possible risk and recurrence factors and propose follow-up strategies for this group of patients. METHODS: A retrospective study from January-2010 to December-2020 analyzed patients with negative conization including all surgical techniques and procedure indications. Biopsy and cervical conizations slides were reviewed and patients who kept a negative result underwent deeper levels sectioning of the paraffin blocks with immunohistochemical stains application: p16, Ki-67 and geminin. Data were compared with a control group composed by 29 women with CIN3. RESULTS: Out of 1022 conizations, 186 were negative (18.1%), with 151 cases selected for the study after excluding 35 patients. Following pathology review, 4 patients were excluded due to false-positive cervical biopsy results, 16 for false-negative conization results and 9 for hidden dysplasia identified after deeper sectioning. The remaining 122 patients were considered truly negative cones (11.9%) and exhibited IHC staining with p16 positive in 20.4% of cases, low Ki-67 expression, and low geminin score in most cases. Specimens with CIN 1 had higher prevalence of p16 staining, Ki-67 expression and geminin score when compared to absence of neoplasia, nevertheless geminin had no statistical difference. Older age, higher parity and IHC pattern with negative p16, low Ki-67 and geminin expressions were identified as risk factors for negative cones (p<0.05). Only 10 patients recurred for high-grade lesions, with no statistically significant risk factors identified. CONCLUSIONS: The negative conization rate was 11.9%, with diagnostic errors identified across pre-surgical biopsy, cone specimen, and deeper levels. Risk factors included older age, higher parity, low expression of p16, Ki-67 and geminin (p<0.05). Recurrence represented 8.1% of the negative cones, without identification of statistically significant risk factors. Pathological review with deeper level sections and 2-year follow-up are recommended for patients with negative conizations.

Reliability of negative cone specimens of the cervix: A review.

OBJECTIVES: Cervical conization specimens with a negative result for high-grade lesion are not infrequent in clinical practice and there are no protocols to address this issue. The purpose of this manuscript is to analyze factors that affect the reliability on these situations and provide recommendations to guide the gynecologists on their practice. METHODS: We searched original articles on Pubmed/Medline database that analyzed negative cones using different combinations of descriptors. There were no restrictions regarding the language or the year of publication. RESULTS: Nineteen articles were selected and a total of 7310 cones analyzed. The negative excision rate ranged from 10 to 35%. Among the reasons to explain absence of lesion, the most frequent were errors in colposcopy, spontaneous lesion regression, complete removal of small lesions during biopsy, errors in the pre-conization material, false-negative results, and excisional error. Pathological specimen review and application of immunohistochemical biomarkers p16 and Ki-67 seemed to improve accuracy and help in challenging differential diagnosis. Recurrence was detected in up to 30%, as seen in positive cones with compromised margins. Importantly, testing for HPV demonstrated benefits in reducing the number of negative cones. CONCLUSIONS: Several factors could contribute to a negative result in a conization. Our main recommendations include: interval of 4-6 weeks between biopsy and conization, repeat the colposcopy during the excision, consider short-term reevaluation for small colposcopy lesions, perform deep sectioning levels in the paraffin block, use of immunohistochemical markers, HPV testing, and strict surveillance during follow-up as performed for positive cases with compromised margins.

Neonatal neutrophils stimulated by group B Streptococcus induce a proinflammatory T-helper cell bias.

BackgroundGroup B Streptococcus (GBS) infection causes inflammatory comorbidities in newborns. While the mechanisms remain unclear, evidence suggests that impaired innate-adaptive immune interactions may be contributory. We hypothesized that GBS-stimulated neonatal neutrophils provide a milieu that may drive proinflammatory T-helper (Th) cell programming.MethodsNeutrophils were stimulated with Type III GBS (COH1); supernatants or intact neutrophils were cocultured with CD4+ T cells or regulatory T cells (Tregs). Resulting intracellular cytokines and nuclear transcription factors were determined by multicolor flow cytometry.ResultsGBS-stimulated neutrophils released soluble mediators that induced greater interleukin-17 (IL-17) responses in neonatal vs. adult CD4+ T cells in the absence of added polarizing cytokines. GBS-stimulated neonatal neutrophils also induced robust expression of the canonical nuclear transcription factors for Th1 (Tbet) and Th17 (IL-17) cells in CD4+ T cells. Following GBS stimulation, both intact neutrophils and neutrophil-derived mediators promoted the generation of Tregs with Th1 and Th17 characteristics.ConclusionGBS-stimulated neonatal neutrophils bias the in vitro Th differentiation program of neonatal CD4+ T cells and promote proinflammatory Th1 and Th17 phenotypes in Tregs. Our data suggest that developmental modifications of innate-adaptive immune cross-talk mechanisms may contribute to the inflammatory complications associated with neonatal GBS infection.