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46. Decrease of in vitro susceptibility of B-CLL cells of patients treated with Viscum album extract towards the applied extract.

Background: Extracts from Viscum album (VA-E) were recognized to induce apoptosis and indirect immuno activation. To exclude possible B-CLL propagating effects, we analysed the in vitro reactions of cultured peripheral blood B-CLL cells from patients receiving VA-E subcutaneously.

Methods: The primary aim of the study was the susceptibility to VA-E induced apoptosis and a suggested proliferation response towards the in vivo applied VA-E. The study has been registered according the German law as an observational study. The patients recruited from medical wards in Germany had a mean age of 66.6 [+ or -] 10.7 years. Only patients which were in contact with the medical wards within the observation period for at least 4 times were taken into account. Thus we had 4 female and 6 male patients. Seven patients had a low staging (Binet A), 2 had Binet B and 1 Binet C; according to the Rai classification, 4 patients were in the low-risk category (Rai 0), 5 in the intermediate-risk category (Rai I-II) and one in the high-risk category (Rai IV). All patients were treated subcutaneously with VA-E twice a week in the morning, i.e. 7 patients with the commercially available whole plant extract HELIXOR[R] P, and 3 patients with HELIXOR[R] A (two of them received HELIXOR[R] P later on too). We measured intracellular expression of apoptosis-associated mitochondrial Apo2.7 molecules and proliferation associated Ki-67 molecules in B-CLL cells from 10 patients treated with VA-E for 12 months by flow cytometry. The cells were incubated for 96h with both VA-E at 0, 10, 100 and 1000 [micro]g/ml, interleukin-6 at 1000 U/ml, and pokeweed mitogen and phytohaemagglutinine at 2.5 [micro]g/ml.

Results: Within the observation period the susceptibility of the B-CLL cells towards the apoptosis-inducing potential of the VA-E significantly decreased, particularly during the last 6 months. Interleukin-6 did not induce apoptosis, but reduced the amount of spontaneous apoptosis of cultured cells. The B-CLL cells were not proliferating per se in the cell cultures; a proliferation response was induced only by the mitogens, but not by interleukin-6. No significant induction of Ki-67 molecules by the VA-E was observed, but an increase of unspecific binding (even in untreated medium controls) during the last months which coincidences with the decrease of mitochondrial Apo2.7 expression. This effect could be due to the physiological induction of blocking anti-ML antibodies in vivo. Due to the progressive nature of the disease, the mean values of peripheral CD[5.sup.+] CD[19.sup.+] B lymphocytes from the patients increased within the observation period, however, statistically not significant (p = 0.09; Wilcoxon)

Conclusions: We can not confirm potential risks in this ex vivo/in vitro setting.

Keywords: Mistletoe extracts; Viscum album; Apoptosis; Proliferation; Leukaemic b cells

A. Bussing (a,b,*), H. Kochskamper (b), S. Rieger (c), J.M. Schierholz (c), D. Schlodder (c)

(a) Chair of Medical Theory and Complementary Medicine, University of Witten/Herdecke, Germany

(b) Krebsforschung Herdecke e.V., Immunological Laboratory, Bochum-Gerthe, Germany

(c) Helixor Heilmittel GmbH & Co. KG Fischermuhle 1, D-72348 Rosenfeld, Germany

*Corresponding author. Tel.: +49 2330 623246; fax: +49 2330 623358.

E-mail address: (A. Bussing).
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Title Annotation:chronic lymphoid leukemia
Author:Bussing, A.; Kochskamper, H.; Rieger, S.; Schierholz, J.M.; Schlodder, D.
Publication:Phytomedicine: International Journal of Phytotherapy & Phytopharmacology
Geographic Code:4EUGE
Date:Oct 1, 2007
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