Comparative Analysis of Multicolor Flow Cytometry and Immunohistochemistry for the Detection of Disseminated Tumor Cells

Abstract

Disseminating cells of a primary solid tumor may represent the origin of metastases and relapses. We aimed at comparing the diagnostic efficacy of multicolor flow cytometry (MFC) and morphology/immunohistochemistry (IHC) in the detection of disseminated tumor cells in the bone marrow (BM) and body fluids of patients with solid tumors, and in pediatric neuroblastoma cases. We investigated 72 samples retrospecively from 50 patients by MFC. Morphology/IHC data were available in 48 cases. In the first cohort, 36 samples derived from 34 patients with various forms of suspected and proven solid tumors and in the second cohort, 36 samples of 16 children with suspected and proven neuroblastoma were analyzed at diagnosis or during follow-up in a 4-color setting by MFC, and the results were compared with those obtained by IHC. In the group of various solid tumors, we found 91% concordance between IHC and MFC, and it was 65% in the neuroblastoma group, and 77% overall. Detection of disseminated tumor cells was found to be more effective by MFC in de novo neuroblastoma samples (100% vs. 86%). The advantage of MFC was even more pronounced when minimal residual disease was evaluated (efficacy, 92% vs. 68%). In contrast, efficacy of IHC was 100% in the group of various solid tumors, whereas it was 91% for MFC. We conclude that MFC and IHC are both essential tools for examining infiltration of BM and body fluids by disseminating solid tumor cells. In the case of neuroblastoma, however, minimal residual disease detection by MFC in a hypoplastic/aplastic BM environment was more effective than IHC, as considerably more cells could be analyzed.

Stratifying nonfunctional pituitary adenomas into two groups distinguished by macrophage subtypes

“…First, CD11b immunohistochemistry on paraffin-embedded sections has been shown to produce variable results based on processing and storage conditions [15, 16]. Second, immunohistochemistry has been argued to be inferior to flow cytometry for analysis of some cell types in tumors [17]. We also found that the percentage of cells in NFPAs that were CD11b + did not vary regionally between the medial versus lateral aspects of the tumor (Figure 1C; Supplementary Figure 2), further validating the choice of this approach.…”

Flow Cytometry Immunophenotyping for Diagnostic Orientation and Classification of Pediatric Cancer Based on the EuroFlow Solid Tumor Orientation Tube (STOT)

“…In this regard, in 2013, we evaluated the (potential) clinical utility of a comprehensive MFC panel of 32 markers for the diagnostic orientation and classification of pediatric solid tumors, based on a limited series of 52 samples from 40 patients suspicious of pediatric cancer [ 22 ]. The panel of markers evaluated in this preliminary study showed high diagnostic accuracy (96%) vs. conventional histopathology [ 22 ], supporting the great potential of MFC to detect tumor cells in pediatric non-Hodgkin (NHL) [ 23 , 24 ] and Hodgkin lymphoma (HL) [ 25 ], RMS [ 26 ], NBL [ 26 , 27 ], and carcinomas [ 28 , 29 ]. In turn, this study also revealed the limited utility and potential redundancy of several markers, together with the need for additional stainings for improved diagnosis of several specific tumor subtypes [ 22 ], that should be combined into a single multicolor antibody combination for screening purposes.…”

Microbial Regulation of Enteric Eosinophils and Its Impact on Tissue Remodeling and Th2 Immunity

“…This is in contrast with the findings by Mishra et al (67), who did not observe significant differences in eosinophils numbers in the GI of SPF and GF mice when quantified by histology. Several factors might have contributed to this discrepancy including the strain of mice (Black Swiss mice vs. C57BL/6 and BALB/c), microbiome differences under SPF conditions, the number of mice utilized in each study (n = 5 vs. n = 12-20) and, lastly, the technique employed for eosinophils quantification, as it is likely that flow cytometry allowed for a more comprehensive and precise quantification than immunohistochemistry using the eosinophil granule protein, major basic protein (MBP) (68). Importantly, we show that intestinal eosinophils in GF mice contain granules of smaller size and less cytoplasmic granule content than SPF controls, which may lead to eosinophil underdetection when using MBP-based methods.…”