EV-TRACK

Search > Results

You searched for: EV100019 (EV-TRACK ID)

Showing 1 - 1 of 1

Results list Study Tree

Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV100019	1/1	Homo sapiens	NAY	DG IAF	Hurwitz MD	2010	38%
Study summary Full title Radiation therapy induces circulating serum Hsp72 in patients with prostate cancer All authors Hurwitz MD, Kaur P, Nagaraja GM, Bausero MA, Manola J, Asea A Journal Radiother Oncol Abstract BACKGROUND AND PURPOSE: Hsp72 found in the extracellular milieu has been shown to play an important (show more...)BACKGROUND AND PURPOSE: Hsp72 found in the extracellular milieu has been shown to play an important role in immune regulation. The impact of common cancer therapies on extracellular release of Hsp72 however, has been to date undefined. MATERIALS AND METHODS: Serum from 13 patients undergoing radiation therapy (XRT) for prostate cancer with or without hormonal therapy (ADT) was measured for levels of circulating serum Hsp72 and pro-inflammatory cytokines (IL-6 and TNF-alpha) using the classical sandwich ELISA technique and the relative expression of CD8(+) T lymphocytes and natural killer (NK) cells was measured using flow cytometry. Mouse orthotopic xenograft of human prostate cancer tumors (DU-145 and PC-3) were used to validate and further characterize the response noted in the clinical setting. The biological significance of tumor released Hsp72 was studied in human dendritic cells (DC) in vitro. RESULTS: Circulating serum Hsp72 levels increased an average of 3.5-fold (median per patient 4.8-fold) with XRT but not with ADT (p=0.0002). Increases in IL-6 (3.3-fold), TNF-alpha (1.8-fold), CD8(+) CTL (2.1-fold) and NK cells (3.2-fold) also occurred. Using PC-3 and DU-145 human prostate cancer xenograft models in mice, we confirmed that XRT induces Hsp72 release primarily from implanted tumors. In vitro studies using supernatant recovered from irradiated human prostate cancer cells point to exosomes containing Hsp72 as a possible stimulator of pro-inflammatory cytokine production and costimulatory molecules expression in human DC. CONCLUSIONS: The current study confirms for the first time in an actual clinical setting elevation of circulating serum Hsp72 with XRT. The accompanying studies in mice and in vitro identify the released exosomes containing Hsp72 as playing a pivotal role in stimulating pro-inflammatory immune responses. These findings, if validated, may lead to new treatment paradigms for common human malignancies. (hide) EV-METRIC 38% (79th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin NAY Focus vesicles Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture DG IAF Protein markers EV: Tubulin/ HSC73 non-EV: Cell organelle protein Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant Separation Method Density gradient Lowest density fraction 1.08g/L Highest density fraction 1.25g/L Orientation Top-down Immunoaffinity capture Selected surface protein(s) anti-human MHC class II Characterization: Protein analysis Western Blot Antibody details provided? No Detected EV-associated proteins Tubulin/ HSC73 Detected contaminants Cell organelle protein ELISA Antibody details provided? No Detected EV-associated proteins Tubulin/ HSC73

				1 - 1 of 1

EV-TRACK ID	EV100019
species	Homo sapiens
sample type	Cell culture
cell type	NAY
condition	NAY
separation protocol	DG IAF
Exp. nr.	1
EV-METRIC %	38