The gut-associated lymphoid tissue represents an integral part of the immune system. Among the powerful players of the mucosa-associated lymphoid tissue are isolated lymphoid structures (ILSs), which as information centers, drive the local (and systemic) adaptive immune responses. Germinal center reactions, taking place within ILSs, involve the coordinated action of various immune cell types with a central role given to B cells. In the current study, we aimed at dissecting the impact of ILSs within non-tumorous colon tissue (NT) on the pathobiology of colorectal cancer (CRC) with metastasis in the liver (CRCLM). In particular, we focused on the immune phenotypes of ILSs and ectopic lymphoid structures (ELSs), built up at matching primary and metastatic tumor sites. We implemented an integrative analysis strategy on the basis of tissue image cytometry and clonality assessment to explore the immune phenotype of ILS/ELS at three tissue entities: NT, CRC, and CRCLM (69 specimens in total). Applying a panel of lineage markers used for immunostaining, we characterized and compared the anatomical features, the cellular composition, the activation, and proliferation status of ILSs and ELSs, and assessed the clinical relevance of staining-derived data sets. Our major discovery was that ILS characteristics at the NT site predefine the immune phenotype of ELSs at CRC and CRCLM. Thereby, B-cell-enriched (CD20) and highly proliferative (Ki67) ILSs and ELSs were found to be associated with improved clinical outcome in terms of survival and enabled patient stratification into risk groups. Moreover, the data revealed a linkage between B-cell clonality at the NT site and the metastatic characteristics of the tumor in the distant liver tissue. Consolidation of immunostaining-based findings with the results of compendium-wide transcriptomic analysis furthermore proposed CD27 as a novel marker of T follicular helper cells within lymphoid structures. Overall, the study nominates the ILS immune phenotype as a novel prognostic marker for patients with metastatic CRC.

Fine needle aspiration cytology (FNAC) is an important diagnostic tool for tumors of the head and neck. However, non-diagnostic or inconclusive results may occur and lead to delay in treatment. The aim of this study was to evaluate the factors that predict a successful FNAC. A retrospective search was performed to identify all patients who received an FNAC at the Department of Otorhinolaryngology, Head and Neck Surgery, Medical University of Vienna. The variables were patients’ age and sex, localization and size of the punctured structure, previous radiotherapy, experience of the head and neck surgeon, experience of the pathologist and the FNAC result. Based on these parameters, a nomogram was subsequently created to predict the probability of accurate diagnosis. After performing 1221 FNACs, the size of the punctured lesion (p = 0.0010), the experience of the surgeon and the pathologist (p = 0.00003) were important factors for a successfully procedure and reliable result. FNACs performed in nodes smaller than 20 mm had a significantly worse diagnostic outcome compared to larger nodes (p = 0.0004). In conclusion, the key factors for a successful FNAC are nodal size and the experience of the head and neck surgeon and the pathologist.

Novel human epidermal growth factor receptor 2 (HER2)-directed therapies have significantly improved outcomes for patients with HER2-positive early-stage breast cancer. Our study assessed the impact of HER2/chromosome enumeration probe 17 ( CEP17 ) ratio on residual cancer burden (RCB) and long-term prognosis following neoadjuvant chemotherapy with dual HER2-targeted therapy using trastuzumab and pertuzumab to identify candidates for chemotherapy de-escalation. Our study included 169 patients with primary invasive HER2-positive breast cancer who received neoadjuvant chemotherapy with trastuzumab and pertuzumab at the Medical University of Vienna from 2014 to 2020. HER2 (ERBB2) gene copy number and HER2/CEP17 ratio were assessed by in situ hybridization. RCB and pathologic complete remission (pCR) served as primary and secondary endpoints, respectively. Univariate and multivariate logistic regression models were applied to analyze associations between outcomes and predictor variables, focusing on the predictive role of HER2/CEP17 ratio, with cutoff values estimated. Progression-free survival (PFS) and overall survival (OS) were estimated by the Kaplan–Meier method. HER2/CEP17 ratio was significantly associated with response to dual-targeted neoadjuvant chemotherapy in primary HER2-positive breast cancer. Optimal HER2/CEP17 cutoff for predicting RCB 0/I was identified at 5.19. HER2/CEP17 ratio was also significantly associated with PFS as a continuous predictor. Multivariate analysis showed that hormone receptor status and the presence of an in situ tumor component significantly influenced therapy response. HER2/CEP17 ratio predicts therapy response and therefore might aid patient stratification for therapy de-escalation with respect to dual neoadjuvant HER2 blockade. Further investigations are warranted to confirm its relevance with other HER2-targeted agents.