Representative immunohistochemical images of APRIL, BAFF, TWEAK and their cognate receptor expression in human gliomas.

<p>All pictures are taken under 20× magnification. APRIL immunoreactivity (A,B) increases from low grade to higher grade lesions. Note the intense staining in larger cells, while smaller ones are negative (A). BAFF immunoreactivity declines from lower grade (C, diffuse and intense), to higher grade gliomas (D). TACI displayed higher immunoreactivity in higher grade gliomas (F) than in low grade ones (E). Similar findings were found for BCMA as well (low grade G, high grade F). Low and high grade tumors displayed heterogeneous regarding percentage of positive cells and/or intensity of staining for TWEAK and Fn14. Here representative photos from Grade III gliomas for TWEAK (I, J) and Fn14 (K, L), respectively are shown.</p

Representative images of BAFF, TWEAK and their cognate receptor BCMA, TACI and Fn14 expression in vascular endothelial cells of human glioma specimens.

<p><b>Upper row:</b> vascular endothelium immunoreactivity for CD31 (<b>A</b>), Fn14 (<b>B</b>), and TWEAK (<b>C</b>). <b>Lower row</b>: vascular endothelium immunoreactivity for BAFF (<b>D</b>) and TACI (<b>E</b>). In panel <b>F</b>, vascular endothelial immunoreactivity for BCMA (direct immunofluorescence assay, with anti-BCMA-FITC antibody. Similar results were obtained by indirect immunostaining with non-fluorescent anti-BCMA antibody (not shown). Arrows show tumor vessels. APRIL was not detected in endothelial cells (not shown). All pictures are taken under 20× magnification. Bar = 200 µm. Inserts in each panel show a higher magnification of a representative part of the vessel.</p

Representative images of BAFF, APRIL, TWEAK and their cognate receptors as well as inflammatory markers CD3, CD20, CD68 expression in human glioma specimens.

<p>Parallel immunostaining of BAFF, APRIL, TWEAK, BCMA, TACI, Fn14 and inflammatory markers CD3, CD20 and CD68 showing that the main source of these TNFSF members are reactive and tumoral glial cells <i>per se</i> in areas with minimal inflammation. Red arrows (black in 3B) indicate gemistocytes immunopositive for BAFF (<b>A</b>), APRIL (<b>B</b>), TWEAK (<b>C</b>), TACI (<b>D</b>), BCMA (<b>E</b>) and Fn14 (<b>F</b>). Gemistocytes negative for lymphocytic CD3 (<b>G</b>), CD20 (<b>H</b>) and panmacrophage CD68 (<b>I</b>) immunostaining. CD68 immunopositive macrophages/microglia and monocytes in I. All pictures are taken under 40× magnification.</p

Correlation between pairs of the TNFSF ligands and receptors identified in our series of 56 gliomas.

<p>The Spearman's correlation coefficient (rho) is reported, together with the number of cases and statistical significance. H-score (as presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083250#pone.0083250.s012" target="_blank">Table S4</a> is used for the analysis.</p><p>*. Correlation is significant at the 0.05 level (1-tailed).</p><p>**. Correlation is significant at the 0.01 level (1-tailed).</p

Quantification of BAFF, APRIL, TWEAK and their receptors in Low (WHO I & II) and high grade gliomas (WHO grade III & IV).

<p>Data are expressed as mean±SEM and medians of the corresponding H-score of tumors, presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083250#pone.0083250.s012" target="_blank">Table S4</a>. Statistical analysis was performed by one way ANOVA, with Bonferroni correction.</p

Quantification of BAFF, APRIL, TWEAK and their receptors in gliomas, stratified by grade.

<p>Data are expressed as mean±SEM and medians of the corresponding H-score of tumors, presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0083250#pone.0083250.s012" target="_blank">Table S4</a> in <b>A</b> and bar graph (mean±SEM) in <b>B</b>.</p

BAFF, APRIL, TWEAK, BCMA, TACI and Fn14 Proteins Are Related to Human Glioma Tumor Grade: Immunohistochemistry and Public Microarray Data Meta-Analysis

<div><p>Gliomas are common and lethal tumors of the central nervous system (CNS). Genetic alterations, inflammatory and angiogenic processes have been identified throughout tumor progression; however, treatment still remains palliative for most cases. Biological research on parameters influencing cell survival, invasion and tumor heterogeneity identified several cytokines interfering in CNS inflammation, oxidative stress and malignant transformation, including TNF-superfamily (TNFSF) members. In this report we performed a meta-analysis of public gene-array data on the expression of a group of TNFSF ligands (BAFF, APRIL, TWEAK) and their receptors (BAFF-R, TACI, BCMA, Fn14) in gliomas. In addition, we investigated by immunohistochemistry (IHC) the tumor cells' expression of these ligands and receptors in a series of 56 gliomas of different grade. We show that in IHC, BAFF and APRIL as well as their cognate receptors (BCMA, TACI) and Fn14 expression correlate with tumor grade. This result was not evidenced in micro-arrays meta-analysis. Finally, we detected for the first time Fn14, BAFF, BCMA and TACI in glioma-related vascular endothelium. Our data, combined with our previous report in glioma cell lines, suggest a role for these receptors and ligands in glioma biology and advance these molecules as potential markers for the classification of these tumors to the proliferative, angiogenic or stem-like molecular subtype.</p></div

Upper Panels.

<p>Predictive value of the retained NT model. Employing a leave-one-out procedure, we attempted to identify whether our models could predict the frequency of emergency department (ED) visits in the metropolitan area of Heraklion. The predictive ability of the NT model, taking into account the ground temperatures (A) was very good, while the SAT model (B), integrating satellite temperature measurements was less performant. Periods refere to half month time-intervals, with the first referring to September 1–15 2007. Lower Panels. Validation of the NT model for severe allergy prediction. In order to validate our NT Model we analyzed a second data-set for the Heraklion metropolitan area (2009–2010) using the same model averaging technique and using the same environmental/meteorological parameters (NDVI, temperature; NT model). Development of the new NT model, based on the same base variables presented a very good model fit both at the Heraklion (C) and at the Chania area (D).</p

Data_Sheet_1_BCMA (TNFRSF17) Induces APRIL and BAFF Mediated Breast Cancer Cell Stemness.docx

<p>Recent advances in cancer immunology revealed immune-related properties of cancer cells as novel promising therapeutic targets. The two TNF superfamily members, APRIL (TNFSF13), and BAFF (TNFSF13B), which are type II membrane proteins, released in active forms by proteolytic cleavage and are primarily involved in B-lymphocyte maturation, have also been associated with tumor growth and aggressiveness in several solid tumors, including breast cancer. In the present work we studied the effect of APRIL and BAFF on epithelial to mesenchymal transition, migration, and stemness of breast cancer cells. Our findings show that both molecules increase epithelial to mesenchymal transition and migratory capacity of breast cancer cells, as well as cancer stem cell numbers, by increasing the expression of pluripotency genes such as ALDH1A1, KLF4, and NANOG. These effects are mediated by their common receptor BCMA (TNFRSF17) and the JNK signaling pathway. Interestingly, transcriptional data analysis from breast cancer cells and patients revealed that androgens can increase APRIL transcription and subsequently, in an autocrine/paracrine manner, enhance its pluripotency effect. In conclusion, our data suggest a possible role of APRIL and BAFF in breast cancer disease progression and provide evidence for a new possible mechanism of therapy resistance, that could be particularly relevant in aromatase inhibitors-treated patients, were local androgen is increased.</p

Graphical presentation of the frequency of emergency department (ED) visits at the Heraklion metropolitan area (solid line) and model fit of the NT Model, that is derived from NDVI and temperature (dotted line) (A) and the SAT Model Assemblage, from NDVI and satellite collected Land Surface Temperature (B).

<p>See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0121475#sec002" target="_blank">Material and Methods</a> for the development of the models. It should be noted that while the raw output of the models is pi, i.e. the proportion of the total population that performed ED visits, results are presented as counts (pi × N), for ease of interpretation in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0121475#pone.0121475.g002" target="_blank">Fig. 2</a> and subsequent Figs. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0121475#pone.0121475.g003" target="_blank">3</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0121475#pone.0121475.g004" target="_blank">4</a>.</p