Simplified diagrammatic representation of our mathematical model.

<p>The dynamic model represents the interaction between healthy urothelial tissue, tumor cells, and their association with BCG (interaction depicted by green line), which in turn results in the generation of BCG-associated healthy tissue and tumor cells. The latter cell populations possess the capacity to trigger the activation of innate immune effector cells and in turn the priming of adaptive immune effector cells. Red lines ending in circles indicate the input stimuli that influence the recruitment of the indicated immune cell populations. Black arrows indicate either increase (e.g., recruitment or proliferation) or decrease (e.g., death or cell turn-over) of the respective cell population. Lines with blunt ends indicate direct killing of target cell populations (solid lines) or bystander death (dashed lines). The model was parameterized using available clinical and <i>in vivo</i> data and tuned to achieve 50% probability of tumor extinction after six weekly intravesical instillations of BCG. A more detailed version of this figure is provided by the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056327#pone.0056327.s001" target="_blank"><b>Figure S1</b></a> of the supplemental material.</p

Timing of BCG therapy, BCG dose and dwell time influence probability of tumor extinction.

<p>(<b>A</b>) Probability of tumor extinction with varying the time from surgery to the start of BCG therapy. The grey shaded area represents the time from surgery to the typical initiation of BCG therapy (i.e., 2 weeks). The cancer growth rate used is based on the clinical observation that high-grade lesions become visible by 3 months in the absence of treatment (detailed definition of tumor dynamics is provided in the supplementary information and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056327#pone.0056327.s002" target="_blank"><b>Figure S2</b></a>). The probability of tumor extinction after six weekly BCG instillations was modeled as a function of (<b>B</b>) BCG dose, and (<b>C</b>) BCG dwell time.</p

The treatment interval strongly influences the probability of tumor extinction.

<p>(<b>A</b>) Schematic representation of the modeled surgery time point followed by intravesical treatment time points (arrows). The symbol * indicates the time of transurethral resection (TUR) followed by the 2 weeks interval before BCG therapy starts. Three weekly instillations are given to initiate innate and adaptive immune responses. The time interval between the third through sixth treatment dose was varied and designated by <i>N</i> (as measured in weeks). (<b>B</b>) Probability of tumor extinction as a function of <i>N</i>, the inter-instillation interval during the 3^rd – 6^th treatment doses. The dotted line marks the outcome after the recommended interval of <i>N</i> = 1 week.</p

Polyclonal rabbit anti GPGRAPI-GST serum: A) After immunization the polyclonal antiserum but not a control protein (scrambled sequence) strongly binds to immobilized synthetic GPGRAPI.

<p>Pre-immune rabbit serum (white column) served as negative control and did not bind to either peptides. B) Binding of the polyclonal antiserum to immobilized GPGRAPI-GST fusion protein is competitively inhibited by synthetic GPGRAPI- but not by the control peptide. Preimmune serum from the same rabbit served as negative control. Error bars represent standard deviation of triplicates, ** indicates p <0.001</p

Ubiquitin is specifically recognized by the immunized rabbit antiserum and the index patient serum as determined by binding inhibition.

<p>A) Addition of 600μg GPGRAPI peptide (p<0,01), but not a control peptide significantly decreases binding of the immunized serum to Ubiquitin. B) Similarly, serum from the index patient (black columns) was competitively inhibited. A human control serum (white columns) bound lower than the index patient serum and was not inhibited by GPGRAPI peptide. Error bars represent standard deviation of triplicates, * indicates p <0.01.</p

Identification of the corresponding antigen to the peptide sequence GPGRAPI.

<p>Protein isolates of the human urothelial cancer bladder cell line T24 was harvested, lysed and fractionated. (N = nuclear, C = cytosolic fraction, M = protein marker). A) Western Blot, stained with preimmune (left gel) and the immunized (right gel) rabbit serum. B) Corresponding gel before (left gel) and after excision (right gel) of the recognized bands for MALDI analysis.</p

ELISA analysis of anti Ubiquitin titers. Left axis depicts absorbance/optical density (O.D.) at 620nm.

<p>A) TCC patients (black columns) have significant higher titers compared to non TCC patients (grey columns) (p<0.001). B) Titers from patients undergoing BCG immunotherapy for high risk TCC do not differ when comparing responder (black columns) to non responder (white columns). Bars represent titer changes against Ubiquitin during BCG-therapy. Inlets depict results of Mann Whitney U test (box and whiskers plots with maximum and minimum range).</p

An Anti-Ubiquitin Antibody Response in Transitional Cell Carcinoma of the Urinary Bladder - Table 1

<p>An Anti-Ubiquitin Antibody Response in Transitional Cell Carcinoma of the Urinary Bladder</p> - Table

An Anti-Ubiquitin Antibody Response in Transitional Cell Carcinoma of the Urinary Bladder - Table 2

<p>An Anti-Ubiquitin Antibody Response in Transitional Cell Carcinoma of the Urinary Bladder</p> - Table

image_1_Expression of Indoleamine 2,3-Dioxygenase Induced by IFN-γ and TNF-α as Potential Biomarker of Prostate Cancer Progression.tiff

<p>Inflammation has been suggested to play an important role in onset and progression of prostate cancer (PCa). Histological analysis of prostatectomy specimens has revealed focal inflammation in early stage lesions of this malignancy. We addressed the role of inflammatory stimuli in the release of PCa-specific, tumor-derived soluble factors (PCa-TDSFs) already reported to be mediators of PCa morbidity, such as indoleamine 2,3-dioxygenase (IDO) and interleukin (IL)-6. Inflammation-driven production and functions of PCa-TDFSs were tested “in vitro” by stimulating established cell lines (CA-HPV-10 and PC3) with IFN-γ or TNF-α. Expression of genes encoding IDO, IL-6, IFN-γ, TNF-α, and their receptors was investigated in tumor tissues of PCa patients undergoing radical prostatectomy, in comparison with benign prostatic hyperplasia (BPH) specimens. IFN-γ and TNF-α-treatment resulted in the induction of IDO and IL-6 gene expression and release in established cell lines, suggesting that the elicitation of PCa-TDSFs by these cytokines might contribute to progression of cancer into an untreatable phenotype. An analysis based on timing of biochemical recurrence revealed the prognostic value of IDO but not IL-6 gene expression in predicting recurrence-free survival in patients (RFS) with PCa. In addition, a urine-based mRNA biomarker study revealed the diagnostic potential of IDO gene expression in urines of men at risk of PCa development.</p