Updated 2013 College of American Pathologists/American Society of Clinical Oncology (CAP/ASCO) guideline recommendations for human epidermal growth factor receptor 2 (HER2) fluorescent in situ hybridization (FISH) testing increase HER2 positive and HER2 equivocal breast cancer cases; retrospective study of HER2 FISH results of 836 invasive breast cancers.

For dual probe HER2 FISH assay, the 2013 CAP/ASCO guideline recommendations lowered the HER2/CEP17 ratio cut off for HER2 amplification to ≥2.0 and introduced an average HER2 copy number criterion for HER2 amplification (≥6.0/cell) and HER2 equivocal categories (≥4 and <6/cell). The HER2/CEP17 equivocal category is eliminated. The aim of this study is to assess the impact of 2013 HER2 FISH testing guideline recommendations update on the assignment of HER2 status with dual probe HER2 FISH assay. Dual probe HER2 FISH assay results on breast cancers from 09/2009 to 07/2015 that underwent reflex HER2 FISH testing after equivocal HER2 (2+) immunohistochemistry (IHC) were reviewed. HER2 copy number, CEP17 signals, and HER2/CEP ratios were noted. HER2 status was assigned as HER2 negative (HER2-), HER2 equivocal (HER2e), and HER2 amplified (HER2+) by applying both 2007 and 2013 CAP/ASCO HER2 FISH guideline recommendations and results were compared. New guidelines reclassified HER2 FISH status in a significant proportion of cases (8.3 %, 69/836; p = .021). There were 22 (2.6 %) more HER2+, 17 (2.1 %) more HER2e, and 39 (4.1 %) fewer HER2- tumors. Change of HER2 status correlated significantly with ≥3 CEP17 signals (38 vs. 2 %; p < .001). The 2013 CAP/ASCO guideline recommendations for HER2 FISH testing by dual probe assay increased the HER2 amplified and HER2 equivocal tumors. Increase in HER2 equivocal tumors would potentially increase the frequency of repeat HER2 testing. Tumors with ≥3 CEP17 signals, so-called chromosome 17 polysomy, are more likely to be impacted and classified as HER2 equivocal.

Application of HER2 CISH pharmDX for DNA Ploidy Determination.

Products of conception (POC) are encountered daily in general pathology practice. The molar workup is an important part of POC examination. Ploidy analysis, expressed as DNA index (DI), is part of the pathologic workup of molar pregnancy. For the past decade, chromogenic in situ hybridization (CISH) has become a popular way to detect HER2 gene amplification. Current study aims to determine whether HER2 CISH dual-color assay can be used to determine DI in POCs. Twenty-two POC cases were chosen from the departmental archives, including 6 complete hydatidiform mole (CM), 10 partial mole (PM), and 6 hydropic POC (HP). CISH assay was performed using the HER2 CISH PharmDx Kit (SK109; Dako). This kit generates red (HER2) and blue (CEN-17) chromogenic signals on the same tissue section. In the 10 triploid PM cases, CISH generated HER2 signal value of 2.925±0.19. Nine cases (90%) had values within this range, except 1 case (2.5). In diploid cases, CISH generated HER2 signal value of 2.063±0.19. Results from 11 (91.7%) cases fell within this range, except 1 HP case (2.35). Sensitivity is 90%, specificity 91.6%, and overall accuracy 90.9%. The current study is the first one that demonstrates HER2/CEN-17 dual-color CISH can be used for microscopic analysis of cell ploidy. This technique provides a relatively easy and straight way to access DI using regular bright-field microscope. Concurrent CEN-17 signal and ploidy in both placental and maternal tissue can be used as internal control. This assay can be performed in any laboratory that can perform immunohistochemistry.

Pulmonary dendritic cells in lungs of preterm infants: neglected participants in bronchopulmonary dysplasia?

Preterm infants are at risk for bronchopulmonary dysplasia (BPD), a chronic lung disease characterized by disrupted alveolar remodeling and microvascular dysangiogenesis. The pathogenesis of BPD is multifactorial, with contributions from antenatal and/or postnatal infection and inflammation. The potential role of dendritic cells, critical immune regulatory cells with potent angiogenic activities, remains undetermined. We studied the prevalence and topography of dendritic cells in postmortem lungs of short- and long-term ventilated preterm infants born between 23 and 29 weeks in gestation. Controls were age-matched infants who had lived less than 12 hours. Dendritic cells were identified by anti-DC-SIGN immunohistochemistry and were co-localized with endothelial and smooth muscle cells by double immunofluorescence. Lungs of early and late control infants without evidence of antenatal infection contained scattered DC-SIGN-positive dendritic cells in the peripheral lung parenchyma. Lungs of early control infants with a history of chorioamnionitis/antenatal infection and lungs of short- or long-term ventilated preterm infants showed a dramatic (more than 3-fold) increase in dendritic cells. Double labeling highlighted a close association between dendritic cells and small- or medium-sized pulmonary vessels. In conclusion, we demonstrated that dendritic cells are an integral component of normal postcanalicular lung development. Antenatal infection and ventilation/BPD are associated with significant pulmonary recruitment of dendritic cells. The recently described angiogenic effects of dendritic cells and their intimate association with the pulmonary microvasculature indicate that dendritic cells may participate in BPD-associated dysangiogenesis. Elucidation of the role of this immunovascular axis may lead to novel therapeutic approaches to BPD.