TY  - JOUR
A1  - Wallaschek, Nina
A1  - Reuter, Saskia
A1  - Silkenat, Sabrina
A1  - Wolf, Katharina
A1  - Niklas, Carolin
A1  - Özge, Kayisoglu
A1  - Aguilar, Carmen
A1  - Wiegering, Armin
A1  - Germer, Christoph-Thomas
A1  - Kircher, Stefan
A1  - Rosenwald, Andreas
A1  - Shannon-Lowe, Claire
A1  - Bartfeld, Sina
T1  - Ephrin receptor A2, the epithelial receptor for Epstein-Barr virus entry, is not available for efficient infection in human gastric organoids
JF  - PLoS Pathogens
N2  - Epstein-Barr virus (EBV) is best known for infection of B cells, in which it usually establishes an asymptomatic lifelong infection, but is also associated with the development of multiple B cell lymphomas. EBV also infects epithelial cells and is associated with all cases of undifferentiated nasopharyngeal carcinoma (NPC). EBV is etiologically linked with at least 8% of gastric cancer (EBVaGC) that comprises a genetically and epigenetically distinct subset of GC. Although we have a very good understanding of B cell entry and lymphomagenesis, the sequence of events leading to EBVaGC remains poorly understood. Recently, ephrin receptor A2 (EPHA2) was proposed as the epithelial cell receptor on human cancer cell lines. Although we confirm some of these results, we demonstrate that EBV does not infect healthy adult stem cell-derived gastric organoids. In matched pairs of normal and cancer-derived organoids from the same patient, EBV only reproducibly infected the cancer organoids. While there was no clear pattern of differential expression between normal and cancer organoids for EPHA2 at the RNA and protein level, the subcellular location of the protein differed markedly. Confocal microscopy showed EPHA2 localization at the cell-cell junctions in primary cells, but not in cancer cell lines. Furthermore, histologic analysis of patient tissue revealed the absence of EBV in healthy epithelium and presence of EBV in epithelial cells from inflamed tissue. These data suggest that the EPHA2 receptor is not accessible to EBV on healthy gastric epithelial cells with intact cell-cell contacts, but either this or another, yet to be identified receptor may become accessible following cellular changes induced by inflammation or transformation, rendering changes in the cellular architecture an essential prerequisite to EBV infection.
KW  - Organoids
KW  - ephitelial cells
KW  - gastrointestinal infections
KW  - cancers and neoplasms
KW  - Epstein-Barr virus
KW  - flow cytometry
KW  - epithelium
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259206
VL  - 17
IS  - 2
ER  - 
TY  - JOUR
A1  - Sander, Brigitta
A1  - de Jong, Daphne
A1  - Rosenwald, Andreas
A1  - Xie, Wanling
A1  - Balagué, Olga
A1  - Calaminici, Maria
A1  - Carreras, Joaquim
A1  - Gaulard, Philippe
A1  - Gribben, John
A1  - Hagenbeek, Anton
A1  - Kersten, Marie José
A1  - Molina, Thierry Jo
A1  - Lee, Abigail
A1  - Montes-Moreno, Santiago
A1  - Ott, German
A1  - Raemaekers, John
A1  - Salles, Gilles
A1  - Sehn, Laurie
A1  - Thorns, Christoph
A1  - Wahlin, Bjorn E.
A1  - Gascoyne, Randy D.
A1  - Weller, Edie
T1  - The reliability of immunohistochemical analysis of the tumor microenvironment in follicular lymphoma: a validation study from the Lunenburg Lymphoma Biomarker Consortium
JF  - Haematologica
N2  - The cellular microenvironment in follicular lymphoma is of biological and clinical importance. Studies on the clinical significance of non-malignant cell populations have generated conflicting results, which may partly be influenced by poor reproducibility in immunohistochemical marker quantification. In this study, the reproducibility of manual scoring and automated microscopy based on a tissue microarray of 25 follicular lymphomas as compared to flow cytometry is evaluated. The agreement between manual scoring and flow cytometry was moderate for CD3, low for CD4, and moderate to high for CD8, with some laboratories scoring closer to the flow cytometry results. Agreement in manual quantification across the 7 laboratories was low to moderate for CD3, CD4, CD8 and FOXP3 frequencies, moderate for CD21, low for MIB1 and CD68, and high for CD10. Manual scoring of the architectural distribution resulted in moderate agreement for CD3, CD4 and CD8, and low agreement for FOXP3 and CD68. Comparing manual scoring to automated microscopy demonstrated that manual scoring increased the variability in the low and high frequency interval with some laboratories showing a better agreement with automated scores. Manual scoring reliably identified rare architectural patterns of T-cell infiltrates. Automated microscopy analyses for T-cell markers by two different instruments were highly reproducible and provided acceptable agreement with flow cytometry. These validation results provide explanations for the heterogeneous findings on the prognostic value of the microenvironment in follicular lymphoma. We recommend a more objective measurement, such as computer-assisted scoring, in future studies of the prognostic impact of microenvironment in follicular lymphoma patients.
KW  - CD/metabolism
KW  - flow cytometry
KW  - antigens
KW  - regulatory T-cells
KW  - independent predictor
KW  - gene expression
KW  - high numbers
KW  - CD40 ligand
KW  - Riutximab
KW  - survival
KW  - marcophages
KW  - transformation
KW  - in-vitro
Y1  - 2014
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-116875
SN  - 1592-8721
VL  - 99
IS  - 4
ER  -