Regulatory T-cells and associated pathways in metastatic renal cell carcinoma (mRCC) patients undergoing DC-vaccination and cytokine-therapy.

PURPOSE: To evaluate CD4(+)CD25(+)FOXP3(+) T regulatory cells (T(REG)) and associated immune-regulatory pathways in peripheral blood lymphocytes (PBL) of metastatic renal cell carcinoma (mRCC) patients and healthy volunteers. We subsequently investigated the effects of immunotherapy on circulating T(REG) combining an extensive phenotype examination, DNA methylation analysis and global transcriptome analysis. DESIGN: Eighteen patients with mRCC and twelve volunteers (controls) were available for analysis. T(REG) phenotype was examined using flow cytometry (FCM). T(REG) were also quantified by analyzing the epigenetic status of the FOXP3 locus using methylation specific PCR. As a third approach, RNA of the PBL was hybridized to Affymetrix GeneChip Human Gene 1.0 ST Arrays and the gene signatures were explored using pathway analysis. RESULTS: We observed higher numbers of T(REG) in pre-treatment PBL of mRCC patients compared to controls. A significant increase in T(REG) was detected in all mRCC patients after the two cycles of immunotherapy. The expansion of T(REG) was significantly higher in non-responders than in responding patients. Methylation specific PCR confirmed the FCM data and circumvented the variability and subjectivity of the FCM method. Gene Set Enrichment Analysis (GSEA) of the microarray data showed significant enrichment of FOXP3 target genes, CTLA-4 and TGF-ß associated pathways in the patient cohort. CONCLUSION: Immune monitoring of the peripheral blood and tumor tissue is important for a wide range of diseases and treatment strategies. Adoption of methodology for quantifying T(REG) with the least variability and subjectivity will enhance the ability to compare and interpret findings across studies.

Passive bioelectrical properties for assessing high- and low-grade prostate adenocarcinoma.

BACKGROUND: The electrical properties of prostate tissues are dependent on cellular morphology and have been demonstrated to distinguish between benign and malignant formations. Because Gleason grading is also based on tissue architecture we explored the hypothesis that the electrical properties might also provide discriminating power between high- and low-Gleason grade cancers. METHODS: Electrical properties (σ, ε, Δσ, σ(∞) , f(c) , and α) were gauged from 546 prostate tissue samples and correlated with histopathological assessment. Primary and secondary Gleason grades and a Gleason score were assigned to the tissues identified as cancer. We evaluated how well differently graded cancers were separable from benign tissues and from each other on the basis of these properties using ROC curves. RESULTS: Of the 546 prostate tissue samples, 71 were identified as cancer and 465 as benign. ε, Δσ, σ(∞) , and f(c) provided the most discriminatory power with area under the curves (AUCs) ranging from 0.77-0.82 for detecting any cancer, 0.72-0.8 for low-grade cancer, and increasing to 0.87-0.9 for detecting high-grade cancer. Further, ε, Δσ, and σ(∞) , provided AUCs ranging from 0.74 to 0.75 for discriminating between low- and high-grade cancers. CONCLUSIONS: Using the electrical properties to identify prostate cancer is improved when high-grade cancers are sought. These electrical properties can also discriminate between different grades of tumors. These findings suggest that technologies being developed to sense and image these properties in vivo may discriminate between aggressive and indolent lesions.

Clinical and immunologic effects of intranodal autologous tumor lysate-dendritic cell vaccine with Aldesleukin (Interleukin 2) and IFN-{alpha}2a therapy in metastatic renal cell carcinoma patients.

PURPOSE: To evaluate the clinical and immunologic outcomes of DC (dendritic cell) vaccine with interleukin (IL)-2 and IFN-alpha 2a in metastatic renal cell carcinoma patients. EXPERIMENTAL DESIGN: Eighteen consented and eligible patients were treated. Peripheral blood monocytes were cultured ex vivo into mature DCs and loaded with autologous tumor lysate. Treatment consisted of five cycles of intranodal vaccination of DCs (1 x 10(7) cells/1 mL Lactated Ringer's solution), 5-day continuous i.v. infusion of IL-2 (18MiU/m2), and three s.c. injections of IFN-alpha 2a (6MiU) every other day. Response Evaluation Criteria in Solid Tumors criteria were used for disease assessment. Correlative immunologic end points included peripheral blood lymphocyte cell phenotype and function as well as peripheral blood anti-renal cell carcinoma antibody and cytokine levels. RESULTS: All patients received between two and five treatment cycles. Toxicities consisted of known and expected cytokine side effects. Overall objective clinical response rate was 50% with three complete responses. Median time to progression for all patients was 8 months, and median survival has not been reached (median follow up of 37+ months). Treatment-related changes in correlative immunologic end points were noted and the level of circulating CD4(+) T regulatory cells had a strong association with outcome. Pre-IP-10 serum levels approached significance for predicting outcome. CONCLUSIONS: The clinical and immunologic responses observed in this trial suggest an interaction between DC vaccination and cytokine therapy. Our data support the hypothesis that modulation of inflammatory, regulatory, and angiogenic pathways are necessary to optimize therapeutic benefit in renal cell carcinoma patients. Further exploration of this approach is warranted.

A da Vinci robot system can make sense for a mature laparoscopic prostatectomy program.

BACKGROUND AND OBJECTIVES: We sought to provide informed recommendations on transitioning from laparoscopic radical prostatectomy (LRP) to robotic-assisted radical prostatectomy (RAP) through a study of the da Vinci robot. METHODS: We performed a cost-benefit analysis to determine the impact that purchasing a dollars 1.5 million da Vinci robot with a dollars 112,000 service contract per year and dollars 200 per case of disposables would have on profits of a mature laparoscopic prostatectomy program. RESULTS: Seventy-eight cases per year are needed to cover the costs of a purchased robot, while only 20 cases per year are needed if a robot is donated. Once robot costs are covered, increases in caseload lead to increased income. Profit is not feasible at centers performing fewer than 25 cases annually. A donated robot lessens costs and allows reasonable revenue without drastic increases in caseload. CONCLUSIONS: Our data suggest a high-volume LRP program can convert to RAP and maintain profits; however, the cost of the robot precludes equal income as that with LRP. Purchasing a robot is not fiscally viable in a low-volume program. Given comparable outcomes between LRP and RAP, hospitals need to decide whether market forces or the intangible benefits of robotics outweigh the expenses of obtaining and operating a robot.

Electrical impedance spectroscopy of the human prostate.

Tissue electrical impedance is a function of its architecture and has been used to differentiate normal and cancer tissues in a variety of organs including breast, cervix, skin, and bladder. This paper investigates the possibility of differentiating normal and malignant prostate tissue using bioimpedance spectra. A probe was designed to measure impedance spectra over the range of 10 kHz to 1 MHz. The probe was fully characterized using discrete loads and saline solutions of different concentrations. Impedance spectra of five ex vivo prostates were measured in the operating room immediately following radical prostatectomy. Wilcoxon signed-rank tests were used to compare the normal and malignant findings. The impedance probe had a signal-to-noise ratio (SNR) > 84 dB across the entire spectrum and measured a tissue volume of approximately 46 mm(3). At 10 kHz, prostate conductivity (or) ranged from 0.232 S/m to 0.310 S/m for tumor and from 0.238 S/m to 0.901 S/m for normal tissue. At 1 MHz the ranges were 0.301 S/m to 0.488 S/m for tumor and 0.337 S/m to 1.149 S/m for normal. Prostate permittivity (epsilonr) ranged from 6.64 x10(4) to 1.25 x 10(5) for tumor and from 9.08 x 10(4) to 4.49 x 10(5) for normal tissues at 10 kHz. And, at 1 MHz the er ranges were 9.23 x 10(2) to 1.88 x 10(3) for tumor and 1.16 x 10(3) to 2.18 x 10(3) for normal tissue. Both sigma and epsilonr of tumor tissue were found to be significantly lower than that of normal tissue (P < 0.0001). Conductivity and permittivity are both higher in normal prostate tissues than they are in malignant tissue making them suitable parameters for tissue differentiation. This is in agreement with trends observed in other tissues reported in much of the literature. Expanded studies are needed to further validate this finding and to explore the biological mechanism responsible for generating the results.

Developing a rational tumor vaccine therapy for renal cell carcinoma: immune yin and yang.

In patients with progressive malignancy, the natural balance between proinflammatory (Yang) and inhibitory (regulatory or Yin) immune pathways is disrupted and favors cancer-specific immune suppression. Therapy with interleukin 2 (IL-2) can mobilize immune effector cells that recognize and destroy cancer. High-dose IL-2 is the only therapy that has consistently induced complete durable remissions in patients with metastatic renal cell carcinoma (RCC) but only in a few of them. The lack of benefit in most metastatic RCC patients is likely due to the ineffective manipulation of other immune circuits critical in regulating tumor cytotoxic pathways. The limited clinical activity of IL-2, RCC vaccines, and other immune therapies to date leads us to postulate that effective clinical treatment strategies will need to simultaneously enhance proinflammatory pathways and disrupt regulatory pathways. We present preliminary studies in RCC patients to highlight the complexity of the regulatory pathways and our approach to shifting the balance of proinflammatory and regulatory immune pathways using dendritic cell-tumor lysate vaccine followed by cytokine therapy.

Tumor-related immunity in prostate cancer patients treated with human recombinant granulocyte monocyte-colony stimulating factor (GM-CSF).

BACKGROUND: Granulocyte monocyte-colony stimulating factor (GM-CSF) supports the survival, expansion, and differentiation of lymphoid and myeloid derived dendritic cells (DCs). We hypothesized that systemic therapy with GM-CSF in prostate cancer patients could augment prostate cancer-related immunity and induce clinical response. METHODS: Eligible patients were randomly assigned to receive either 125 or 250 microg/m(2) GM-CSF subcutaneously three times a week until clinical progression. Prostate-specific antigen (PSA) T cell precursor frequencies were determined by a flow cytometric method. RESULTS: We were able to show, for the first time, a statistically significant correlation between pre-treatment PSA level and PSA-specific CD4(+) T cell precursors and a trend between pre-treatment PSA level and PSA-specific CD8(+) T cell precursors (P<0.0001 and P=0.059, respectively). CONCLUSIONS: These results suggest that existent immunity to PSA in prostate cancer patients may be a promising target for future immunotherapeutic approaches to prostate cancer.

Impaired cytolytic activity in peripheral blood T cells from renal cell carcinoma patients.

Classic T lymphocyte cytotoxicity is mediated through the T cell receptor (TCR). Defects in TCR signal transduction and cytolytic activity have been reported in tumor infiltrating T lymphocytes. We hypothesized that impaired cytotoxicity occurs in peripheral blood T cells from renal cell carcinoma (RCC) that can be reversed by exposure to rhIL-2. Peripheral blood mononuclear cells (PBMC) from 29 RCC patients and 29 healthy volunteers were isolated and cultured in the absence or presence of 10 IU/ml rhIL-2. A redirected cytotoxicity assay that requires TCR signal transduction was used with chromium-labeled P815 target cells, effector PBMC and anti-CD3 antibody. Target cell lysis was measured in "lytic units" (LU). Mean LU from RCC patients was lower than that of healthy volunteers (105.8 LU vs. 194.6 LU, P = 0.025). Exposure to rhIL-2 increased T cell-mediated lysis in both groups. Disruption of T cell cytotoxicity in RCC patients can be overcome by exposure to rhIL-2.

Occupation and bladder cancer risk in a population-based case-control study in New Hampshire.

OBJECTIVE: To identify occupations with excess bladder cancer risk in New Hampshire, where bladder cancer mortality rates have been elevated for decades. METHODS: Lifetime occupational histories were obtained from interviews with 424 cases and 645 controls in a population-based case-control study. Unconditional logistic regression models were used to estimate odds ratios (Ors) and 95% confidence intervals (CI) for each occupation, adjusted for age and smoking. Analyses by duration of employment were carried out and interactions with smoking were examined. RESULTS: Male tractor-trailer truck drivers had an elevated risk for bladder cancer (OR = 2.4, CI = 1.4-4.1), with a significant positive trend in risk with increasing duration of employment (P (trend) = 0.0003). Male metal/plastic processing machine operators also had a significant excess (OR = 4.9, CI = 1.6-15.1), attributable mainly to molding/casting machine operators (OR = 16.6, CI = 2.1-131). Elevated risk was also observed for male fabricators, assemblers, and hand workers (OR = 1.8, CI = 1.0-3.4). Women in certain sales occupations (sales clerks, counter clerks, and cashiers) had a significant excess risk (OR = 2.2, CI = 1.3-3.9) and a significant trend with duration of employment (P (trend) = 0.016), as did female health service workers (OR = 4.1, CI = 1.6-10.7; P (trend) = 0.014). There was a positive interaction between smoking and employment as a health service worker (p = 0.036). CONCLUSIONS: These findings are generally consistent with previous studies. Elevated risks for male molding/casting machine operators, female salesworkers, and female health service workers, especially those with a history of smoking, require further investigation.

Bladder cancer risk and personal hair dye use.

Several cohort and case-control studies have found an increased risk of bladder cancer among hairdressers and barbers who are occupationally exposed to hair dyes. However, the carcinogenic risk associated with personal use of hair dyes remains uncertain since several large case-control and cohort studies did not find an association between personal hair dye use and bladder cancer. To address this question, the authors used data collected on 459 bladder cancer cases and 665 controls who were interviewed as part of a case-control study conducted in New Hampshire between 1994 and 1998. Participants underwent a structured personal interview with regard to history of hair dye use and bladder cancer risk factors. Unconditional logistic regression analysis was used to compute odds ratios that were associated with hair dye use, while controlling for potential confounding factors. A history of any hair dye use was inversely associated with bladder cancer incidence in men [adjusted odds ratio (OR) = 0.5; 95% confidence interval (CI)=0.3-0.8], although risk reductions were not statistically significant for individual dye types. In women, use of permanent (adjusted OR = 1.5; 95%CI = 0.8-2.7) and rinse-type hair dye (adjusted OR = 1.7; 95%CI = 0.8-3.6) were associated with a modestly elevated risk of bladder cancer but with limited statistical precision; no association was found with use of semi-permanent dyes (adjusted OR = 0.7; 95%CI = 0.3-1.4). For permanent hair dye use, odds ratios were most pronounced for younger age at first use, higher frequency and prolonged time since first use; however there were no clear trends in risk by these factors. In light of the prevalence of hair dye use, further studies are needed that address the effects of specific colors and types of hair dyes along with the possible role of individual susceptibility.

Immunological effects of granulocyte-macrophage colony-stimulating factor and autologous tumor vaccine in patients with renal cell carcinoma.

PURPOSE: Biological therapy for renal cell carcinoma (RCC) uses agents that mobilize immune effector cells which are able to recognize and destroy cancer. We evaluated the effects of weekly then monthly autologous tumor vaccine combined with daily granulocyte macrophage-colony stimulating factor (GM-CSF) in patients with RCC as a method of stimulating antigen presenting cells. MATERIALS AND METHODS: Eligible patients with pathological stage II to IV RCC were entered into this pilot study. Autologous tumor vaccine (0.5 to 1 x 107 irradiated tumor cells) admixed with 250 microg GM-CSF per vaccine was given subcutaneously weekly for 4 weeks and then monthly for 4 months. GM-CSF (125 microg/m2) was given subcutaneously for 13 days after vaccine injection 1 and injections 4 to 8. Treatment related tumor specific CD4 and CD8 positive T cell precursors were assessed. RESULTS: A total of 22 patients were entered into this study. Patients were stratified by bulk of disease (group 1, 9 patients with micrometastatic disease, and group 2, 13 patients with macrometastatic disease). In general treatment was well tolerated. Of 9 patients in group 1 7 remained disease-free after nephrectomy. In group 2, 6 patients had stable (46.2%) and 7 patients had progressive disease (53.8%). Statistically significant treatment related increases in CD4 (p = 0.028) and CD8 (p = 0.018) positive tumor specific T cell precursors were observed for the entire group of patients. Changes in CD4 and CD8 positive precursors correlated significantly with each other (p = 0.0001). This correlation was seen in the 2 patient subpopulations as well (group 1 p = 0.003, group 2 p = 0.013). Patients with minimal disease, and with changes in CD4 and CD8 positive tumor specific T cell precursors greater than the median appeared to have an improved time to progression as well as a survival benefit. CONCLUSIONS: GM-CSF and autologous vaccine can be given safely in combination to patients with renal cell cancer. We observed treatment related changes in tumor specific circulating lymphocyte populations.