Chromosomale Imbalancen als mögliche Prognosefaktoren für maligne Mesotheliome?

Basierend auf molekularpathologischen Ergebnissen von 90, mit der komparativen genomischen Hybridisierung (CGH) untersuchten Malignen Mesotheliomen (MM), wurde geprüft, ob sich die erhobenen molekularen Befunde mit klinischen Krankheitsverläufen korrelieren lassen. Hierzu wurde ein Fragebogen mit insgesamt 32 Parametern an die 42 bundesweit lokalisierten behandelnden Kliniken übersandt. Verstorbene Patienten mit Tumoren ohne mit der CGH nachweisbare Defekte zeigten eine längere durchschnittliche ÜLZ als mit Defekten. Verstorbene Patienten mit Veränderungen auf Chromosom 7 wiesen eine kürzere ÜLZ auf. Bei MM mit kurzen ÜLZ (<6 Monate) zeigten sich in je 60% Verluste auf Chromosom 4 und Zugewinne auf Chromosom 7. Die beste Prognose erreichen MM mit fehlendem Nachweis chromosomaler Imbalancen bei der CGH

Cognitive factors mediate placebo responses in patients with house dust mite allergy.

BACKGROUND: Placebo effects have been reported in type I allergic reactions. However the neuropsychological mechanisms steering placebo responses in allergies are largely unknown. The study analyzed whether and to what extend a conditioned placebo response is affecting type I allergic reactions and whether this response can be reproduced at multiple occasions. METHODS: 62 patients with house dust mite allergy were randomly allocated to either a conditioned (n = 25), sham-conditioned (n = 25) or natural history (n = 12) group. During the learning phase (acquisition), patients in the conditioned group received the H1-receptor antagonist desloratadine (5mg) (unconditioned stimulus/US) together with a novel tasting gustatory stimulus (conditioned stimulus/CS). Patients in the sham-conditioned control group received the CS together with a placebo pill. After a wash out time of 9 days patients in the conditioned and sham-conditioned group received placebo pills together with the CS during evocation. Allergic responses documented by wheal size after skin prick test and symptom scores after nasal provocation were analyzed at baseline, after last desloratadine treatment and after the 1(st) and 5(th) CS re-exposure. RESULTS: Both conditioned and sham-conditioned patients showed significantly decreased wheal sizes after the 1(st) CS-evocation and significantly decreased symptom scores after the 1(st) as well as after the 5(th) evocation compared to the natural history control group. CONCLUSIONS: These results indicate that placebo responses in type I allergy are not primarily mediated by learning processes, but seemed to be induced by cognitive factors such as patients' expectation, with these effects not restricted to a single evocation

Modulation and Apoptosis of Neutrophil Granulocytes by Extracorporeal Photopheresis in the Treatment of Chronic Graft-Versus-Host Disease

<div><p>Chronic graft-versus-host disease (cGVHD) is a common side effect of allogeneic stem cell transplantation and a major cause of morbidity and mortality in affected patients. Especially skin, eyes and oral mucosa are affected. This can lead to pain and functional impairment. Extracorporeal photopheresis (ECP) is an effective immunomodulatory therapy with minimal side effects but its mode of action is still largely unknown. The objective of the present study was to examine the effects of ECP on neutrophil granulocytes in patients with cGVHD. Analysis of leukocytes from cGVHD patients obtained from the ECP device during treatment showed that neutrophil granulocytes account for the majority of cells treated during ECP. Neutrophils from healthy donors treated <i>in vitro</i> with 8-methoxypsoralen and UVA light as well as neutrophils from buffy coats of patients with cGVHD treated by ECP showed increased apoptosis and decreased half-life. In remaining non-apoptotic cells chemoirradiation resulted in loss of activation markers and reduced effector functions. This was accompanied by an increase in extracellular arginase-1 activity. Additional comparison of neutrophils isolated from blood of cGVHD patients before and 24h after ECP revealed a decreased half-life and reduction of effector functions of post-ECP neutrophils <i>ex vivo</i>. These observations strongly suggest that ECP induces both apoptosis and physiological changes in neutrophils and that these changes also take place <i>in vivo</i>. This study is the first to show that ECP modulates apoptosis and inflammatory activity in neutrophil granulocytes, indicating that neutrophils may significantly contribute to the overall immunomodulatory effects attributed to this treatment.</p></div

Modulation and Apoptosis of Neutrophil Granulocytes by Extracorporeal Photopheresis in the Treatment of Chronic Graft-Versus-Host Disease - Fig 2

<p><b>(A)</b><i>In vitro</i> treatment of healthy donor neutrophils with 8-methoxypsoralen and UVA induces apoptosis and decreases activation markers. Neutrophils, isolated from blood of healthy donors, were treated with 8-MOP and UVA light and cultured for 24h. Control groups were untreated neutrophils, neutrophils treated with 8-MOP and neutrophils treated with UVA light. Percentage of non-apoptotic cells was determined by the % of Annexin V and 7-AAD double-negative cells via flow cytometry 0, 4, 8 and 24h after irradiation (n = 4). Curves show mean ± standard deviation (SD). ** p≤0.01. <b>(B-D)</b> Viable (= Annexin^- / 7-AAD^-) neutrophils of the same groups (n = 4) were stained for activation markers CD11b, CD16 and CD54 at 2, 4, 8 and 24h after irradiation and protein expression was measured by flow cytometry after gating on viable cells. Curves show mean of Δ median of MFI ± SD. MFI = mean fluorescence intensity, Δ median of MFI = MFI of antibody-MFI of isotype shown.</p

Reduced activation and increased extracellular arginase-1 activity in chemoirradiated neutrophils from buffy coat of cGVHD patients.

<p>(A) Neutrophil granulocytes and mononuclear cells substituted with 8-methoxypsoralen (8-MOP) were obtained from the buffy coat of patients with cGVHD treated with a Therakos UVAR XTS device before (pre-UVA) and after (post-UVA) irradiation. (B) Neutrophils of the buffy coat before and after irradiation with UVA (as described in A) were stained with Annexin V and 7-AAD. Fold change of Annexin V^-/7-AAD^-double negative (viable) cells determined after 24h of ex vivo culture after treatment normalized to cells assessed directly after treatment (0h). Data show means of 3 patients ± standard deviation (SD). **p≤0.01. (C) Neutrophils from buffy coats of cGVHD patients (n = 3) were stained with antibodies against activation markers CD11b, CD16 and CD54 after 24h in culture. Expression was measured by flow cytometry after gating on viable cells. Data shown as difference of the median of the mean fluorescence intensity (mfi) of the specific antibody and the isotype. (D) Supernatants of neutrophils of the samples described in (A) were generated with and without stimulation with LPS for 24h and concentration of CXCL8 and CCL4 was quantified by ELISA. Mean values ± standard deviation (SD) of 3 patients in pg/ml shown. (E) Arginase activity in the same supernatants as in (D) was determined by enzymatic activity (n = 3). Values shown in U/L. Unit definition: 1 unit of arginase converts 1μmol of L-arginine to ornithine and urea per minute at 37°C and pH 9.5. *p≤0.05</p

Neutrophils account for the majority of cells treated during ECP.

<p>(A) Percentages of neutrophils, lymphocytes and monocytes of total leukocytes in buffy coats of 16 patients with chronic GVHD (cGVHD) taken from Therakos UVAR XTS ECP- device immediately after chemoirradiation prior to reinfusion. Lines show means; scatters represent individual patients. **p≤0.01, ****p≤0.0001. (B) Representative composition of a buffy coat of one patient with cGVHD.</p

<i>In vitro</i> treatment with 8-methoxypsoralen and UVA reduces pro-inflammatory effector functions in healthy donor neutrophils.

<p>(A) Neutrophil granulocytes of healthy donors were stimulated with phorbol 12-myristate13-acetate (PMA) for 0, 4, 8 and 24h after treatment with 8-methoxypsoralen and UVA light. Nitric oxide was quantified via flow cytometry by staining with DAF-FM Diacetate after gating on viable cells. Data shown from one representative donor. (B) After 24h of stimulation with LPS, release of CXCL8 and CCL4 into the supernatants was detected by ELISA. Concentrations are shown as x-fold change compared to unstimulated neutrophils. Mean changes of 6 donors ± standard deviation (SD) shown. **p≤0.01. (C) Supernatants of untreated neutrophils and neutrophils treated with 8-MOP and UVA were generated after 24h stimulation with LPS, TNFα and IFNγ (n = 6). Arginase activity was determined enzymatically. Unit definition: 1 unit of arginase converts 1μmol of L-arginine to ornithine and urea per minute at 37°C and pH 9.5. *p = 0.04</p

Experimental design.

<p>During acquisition, patients in the conditioned group received desloratadine (US) in combination with the CS. During evocation, the drug was replaced by placebo pills. Sham-conditioned patients received the CS together with placebo pills throughout the study. During 3 subsequent days during acquisition (2–4) and evocation (17–19) patients were instructed to intake the pills together with the drink (CS) at home.</p