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PubMed1989

Modulatory potency of the beta-galactoside-specific lectin from mistletoe extract (Iscador) on the host defense system in vivo in rabbits and patients.

Proprietary extract of mistletoe (Iscador) that has federal approval for clinical application can exhibit immunomodulatory capacity. However, the nature of this responsible substance has still remained elusive. To validate the hypothesis that specific lectin-carbohydrate interactions at least participate in eliciting immunomodulation, the modulatory efficiency of the major beta-galactoside-specific mistletoe lectin (ML I) from the clinically applied extract on selected immunological parameters was monitored "in vivo" in rabbits. Injections of nontoxic doses of the purified lectin or even only of its carbohydrate-binding subunit (0.25-1.0 ng/kg) into rabbits yielded significant increases in natural killer cytotoxicity, frequency of large granular lymphocytes, and phagocytic activity of granulocytes. In the clinically relevant situation, changes in these parameters were also determined in cancer patients after extract (Iscador) injection s.c. as well as i.v., emphasizing the potential relevance of the lectin. Comparative analyses of the changes in the selected parameters following injection of extract with normal lectin content as well as of extract, selectively depleted of lectin, into healthy volunteers corroborated this inference. Lectin depletion by affinity chromatography was highly specific and did not affect any other substance in the extract. Remarkably, contamination by endotoxin has been rigorously excluded in each applied specimen. These results encourage detailed elucidation of lectin action on various parts of the tumor defense system "in vitro" with the long range goal of achieving progress in the treatment of cancer through immunological strategies, exploring selective mediatory lectin-carbohydrate interactions.

Adjuvants, Immunologic↗
PubMed1982

Isolation and characterization of viscumin, a toxic lectin from Viscum album L. (mistletoe).

A toxic protein, viscumin, was isolated from extracts of mistletoe by affinity chromatography on acid-treated Sepharose 4B. Viscumin was selectively bound to the column and could be eluted with lactose. It migrated in polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate corresponding to Mr = 60,000. In addition, two bands migrating corresponding to Mr = 29,000 and 32,000 were found. After treatment with 2-mercaptoethanol, only 2 bands (Mr = 29,000 and 34,000) were found. Apparently, viscumin consists of two chains which, in some of the molecules, are disulfide-linked. Protection experiments with antiserum against viscumin indicated that the major part of the cytotoxic activity in mistletoe extracts is due to viscumin. Gel filtration experiments on Sephacryl 200 indicated that, at low concentrations, viscumin occurs as a monomer and at higher concentrations as a dimer. Viscumin was found to inhibit protein synthesis in cell-free systems. When the two constituent peptide chains of viscumin were eluted from polyacrylamide gels and tested for ability to inhibit cell-free protein synthesis, this property was found to be associated with the fastest migrating chain, here denoted the A chain. The heavier chain was denoted the B chain. The A chain was found to inhibit protein synthesis by inactivating the ribosomes catalytically. Reconstitution experiments with isolated ribosomal subunits from untreated and A chain-treated ribosomes showed that the 60 S ribosomal subunit was selectively inactivated.

Animals↗
PubMedDate not supplied

Immunoprotective activity of the galactoside-specific mistletoe lectin in cortisone-treated BALB/c-mice.

Hydrocortisone-acetate (HA)-treatment of BALB/c-mice induced a profound suppression of the lymphatic immune system with statistically significant decreases of thymocyte proliferation and maturation rates as well as peripheral blood lymphocyte (PBL) counts. To check its putative immunoprotective/immunorestoring activity, the optimal immunomodulating dosage of commercially available mistletoe extract standardized for the galactoside-specific mistletoe lectin (ML-1) was regularly administered (1 ng ML-1 per kg body weight; subcutaneously). As compared to counts of thymic lymphatic subsets of HA-treated mice, a considerable up regulation of mature cells expressing helper/inducer (L3T4+) as well as cytotoxic/suppressor (Lyt-2+) phenotypes and immature cells expressing both antigens (L3T4+/Lyt-2+) could be found after ML-1 co-administration. Counts of BALB/c-mouse peripheral blood mononuclear cells also revealed statistically significant increases after ML-1 co-administration, as compared to HA-treated animals. Accordingly, ML-1 treatment may be supposed to restore the lymphatic system after immunosuppressive corticoid treatment and thus this treatment may be of great benefit to patients.

Adjuvants, Immunologic↗
PubMedDate not supplied

Thymocyte proliferation and maturation in response to galactoside-specific mistletoe lectin-1.

Commercially available mistletoe extract standardized for the galactoside-specific mistletoe lectin-1 (ML-1) could be shown to induce thymocyte proliferation and maturation in BALB/c-mice after regular subcutaneous administration of the optimal immunomodulating dosage (1ng lectin/kg body weight). The increase in thymocyte numbers per mg organ weight was statistically significant. Determinations of thymic lymphatic subsets revealed considerable up-regulation of mature cells expressing helper/inducer (L3T4) or cytotoxic/suppressor (Lyt-2) phenotype and immature cells presenting both (L3T4/Lyt-2) antigens. Thus, administration of ML-1 accelerated murine thymocyte proliferation and maturation. Counts of BALB/c-mouse peripheral blood lymphocytes (PBL) revealed evident (but statistically non-significant) increases after ML-1 administration. The determination of activated PBL expressing interleukin (Il)-2 receptors proved, however, that ML-1 induced a potent immunostimulation since these cells were significantly enhanced after ML-1 administration.

Adjuvants, Immunologic↗
PubMed1996

[The mistletoe myth--claims, reality and provable perspectives].

Intuition guided R. Steiner to refer to mistletoe as the future remedy for cancer. He proposed that its spiritual qualities support re-establishment of the harmonious integration of the alleged four different entities of human existence in a patient. The assumption of potency without chemical basis is derived from the dogmatic system of anthroposophic reasoning. It explains the evidently similar claim of clinical efficiency for the proprietary mistletoe extracts despite the lack of information on the actual contents of the complex mixtures and despite the conspicuous diversity of methods of manufacture for these products. Thorough scientific analysis of the published clinical experience does not justify this claim. Due to the increasing reference to defined substances in advertisements for commercial extracts, they should no longer avoid rigorous testing according to common quality standards. Interdisciplinary research efforts on the immunomodulatory galactoside-binding lectin illustrate how to yield a clinically testable substance from an ill-defined extract, thereby providing a notable example for rational investigation of unconventional treatment modalities.

Adjuvants, Immunologic↗
PubMed1995

Immunomodulating ability of galactoside-specific lectin standardized and depleted mistletoe extract.

Commercially available mistletoe extract standardized for the galactoside-specific lectin (ML-1; Eurixor) and a chromatographically ML-1-depleted preparation (same charge no. and composition of remaining components) were tested for their immunomodulating potency. In BALB/c-mice, regular subcutaneous administration of the optimal immunomodulating dosage (1 ng ML-1/kg body weight) could be shown to induce no influence on spleen weight, a non-significant increase of thymus weight, a significant increase of thymocyte, peritoneal macrophage, peripheral blood leukocyte, lymphocyte and monocyte and monocyte counts, and a significant decrease of peripheral blood granulocyte counts. Administration of analogue volumes (concentrations) of ML-1-depleted extract, however, did not induce any immunopotentiation. Accordingly, it may be assumed, that the galactoside-specific lectin (ML-1) represents the main immunomodulating component in commercially available mistletoe extracts.

Adjuvants, Immunologic↗
PubMedDate not supplied

Immunomodulating efficacy of combined administration of galactoside-specific lectin standardized mistletoe extract and sodium selenite in BALB/c-mice.

The immunomodulating abilities of commercially available mistletoe extract standardized for the galactoside-specific lectin (mistletoe lectin-1, ML-1) and sodium selenite (Se) were evaluated in BALB/c-mice and compared to non-treated control animals. Following the optimal schedule of administration (ML-1: 1ng/kg BW; days 1, 2, 3, 5 and Se: 3,5 micrograms/kg BW for 7 subsequent days before evaluation) yielded enhanced counts (thymocytes; peritoneal macrophages, MO; peripheral blood leukocytes; lymphocytes, PBL; monocytes, PBM) and activities (CD-25 positive PBL, MAC-3 positive PBM) of desired immune cells reaching statistical significance for most parameters. However, combined administration of ML-1 and Se proved to be superior to monotherapy since immune cell counts (thymocytes, leukocytes, PBL, PBM) and activities (CD-25 positive PBL) exceeded values obtained after monotherapy. These data are in favour of therapeutical strategies in complementary oncology and suggest that the combination of defined immunomodulating substances might positively enhance the efficacy.

Animals↗
PubMed1992

Analysis of tumour necrosis factor alpha-specific, lactose-specific and mistletoe lectin-specific binding sites in human lung carcinomas by labelled ligands.

Receptor sites can be visualized by labelled ligands as an alternative to receptor-specific antibodies, as substantiated for two different receptor classes. Recombinant tumour necrosis factor alpha (TNF) was biotinylated via amino-groups and the resultant probe was applied to formalin-fixed, paraffin-embedded tissue sections of 94 primary bronchial carcinomas and to normal peripheral lung parenchyma. In addition, monoclonal antibodies specific for neuron-specific enolase (NSE) and TNF itself were used. The biotinylated beta-galactoside-specific mistletoe lectin, which exhibits dose-dependent immunomodulatory and toxic potency, and two probes that specifically detect certain types of sugar receptors were employed to illustrate further the feasibility of using ligands for receptor localisation. The tumours comprised 62 small cell lung carcinomas, 10 epidermoid carcinomas, 11 adenocarcinomas and 11 large cell anaplastic carcinomas. Expression of TNF-binding sites was found in 39 of the small cell lung carcinomas and in 13 of the non-small cell lung carcinomas. Binding capacity for the TNF-specific antibody was seen in similar proportions of small cell lung carcinomas and of non-small cell lung carcinomas. None of the normal lung parenchymas revealed significant staining. Binding capacities to mistletoe lectin were seen in all normal lung parenchymas and in nearly all cases of adenocarcinoma (10/11). A correlation between the expression of NSE and the binding capacities to TNF was detected. Endogenous lectins, specific for lactose or beta-GalNAc, were displayed in nearly one half of the small cell lung carcinoma cases (44% or 45% respectively) and in about 25% of the non-small cell lung carcinoma cases.

Acetylgalactosamine↗
PubMed2002

Measuring positive, negative, and null impacts of forest disturbances: a case study using dwarf mistletoe on Douglas fir.

Not all disease activity causes an impact. Not all impacts are negative. The aim of this study was to examine a method that could conceptually specify when impacts occur and that could quantify both negative and positive disease impacts. For this study, dwarf mistletoe ( Arceuthobium douglasi) of Douglas fir ( Pseudotsuga menziesii) in southwestern Oregon was used as a case study. The method uses six variables for forest growth, mortality, and stand structure, and six categorical disease severity scores. The impact model displays stands as points in multidimensional scaling space, where relative position is determined by values of the six stand variables. Positions in this two-dimensional space change when stand characteristics change. Differences associated with disease severity could be traced as trajectories, and impact was quantified using the length and direction of these trajectories. This multivariate impact assessment method was contrasted to impact assessments based on single variables. Methods based on multiple variables offer a useful way of characterizing impact on multiple objectives. The model indicates that dwarf mistletoe has positive, negative, and neutral impacts and that these could be illustrated and quantified using this method.

Models, Theoretical↗
PubMed1995

Native and/or asialo-Tamm-Horsfall glycoproteins Sd(a+) are important receptors for Triticum vulgaris (wheat germ) agglutinin and for three toxic lectins (abrin-a, ricin and mistletoe toxic lectin-I).

The binding properties of human Tamm-Horsfall Sd(a+) urinary glycoprotein (THGP) and asialo-THGP with Triticum vulgaris agglutinin(WGA) and three toxic lectins (abrin-a, ricin, and Mistletoe toxic lectin-I) were investigated by quantitative precipitin and precipitin inhibition assays. Both glycoproteins reacted strongly with abrin-a, precipitating over 80% of the lectin nitrogen tested. THGP also bound well to mistletoe toxic lectin-I and precipitated 86% of this lectin added, while the precipitability of its asialo product decreased by 28%. The native glycoprotein completely precipitated the WGA added, but its reactivity was reduced dramatically after desialylation. On the contrary, the poor reactivity of THGP with ricin increased substantially after removal of sialic acid and completely precipitated the lectin added. The glycoprotein-lectin interactions were inhibited by one or several of the following haptens, p-NO2-phenyl alpha GalNAc, p-NO2-phenyl beta GalNAc, Gal beta 1-->4GlcNAc, Gal beta 1-->4Glc, GlcNac beta 1-->4GlcNAc and/or GlcNAc. From the above results, it is concluded that native and/or Tamm-Horsfall glycoproteins serve as important receptors for these three toxic lectins and for WGA.

Abrin↗
PubMed1992

Carbohydrate specificity of the receptor sites of mistletoe toxic lectin-I.

The carbohydrate specificity of mistletoe toxic lectin-I (ML-I) was studied by haemagglutination-inhibition assay. The results indicated that ML-I has a broad range of affinity for Gal alpha,beta linked sequences. The galabiose (E, Gal alpha 1----4Gal) sequence, a receptor of the uropathogenic E. coli ligand, was one of the best disaccharide inhibitors tested. The lectin also exhibits affinity for Lac(Gal beta 1----4Glc), T(Gal beta 1----3GalNAc), I/II(Gal beta 1----3/4GlcNAc) and B(Gal alpha 1----3Gal) sequences. Gal alpha 1----4Gal and Gal beta 1----4Glc are frequently occurring sequences of many glycosphingolipids located at the mammalian cell membranes, such as intestinal and red blood cell membranes, for ligand binding and toxin attachment. This finding provides important information concerning the possible mechanism of intoxication of cells by the mistletoe preparation.

Binding Sites↗
PubMed1996

Mistletoe lectins I, II and III induce the production of cytokines by cultured human monocytes.

The three mistletoe (Viscum album L.) lectins. ML I, ML II and ML III, were tested on their ability to enhance the secretion of the cytokines tumor necrosis factor (TNF)alpha, interleukin (IL)-1 alpha, IL-1 beta and IL-6 by human monocytes obtained from healthy donors. At lectin concentrations from 0.02 to 20/pg ml (100-10,000-fold lower than those showing toxic effects), stimulations of cytokine production several-fold over control values were observed. The immunoactivating concentrations by the three lectins were found different for each donor. At toxic concentrations, the amounts of IL-1 alpha, IL-1 beta and to a less extent of TNF alpha in monocytes supernatants were particularly high. The data are discussed in relationship with the cytotoxic and immunoactivating effects of mistletoe lectins and their interest in cancer treatment.

Cells, Cultured↗
PubMed2006

Investigation of the effect of mistletoe (Viscum album L.) extract Iscador on the proliferation and apoptosis of murine thymocytes.

Mistletoe (Viscum album L.) extracts (ME) have been shown to exhibit a bell-shaped curve of immunological efficacy and mistletoe lectins (MLs) were found to play an important role in this phenomenon. The aim of present in vivo study was to investigate the acute- and long-term effect of a standardized ME (Iscador M special) on thymocyte subpopulations and peripheral T cells using a murine (Balb/c) model. In thymus CD4-CD8- double negative (DN), CD4+CD8+ double positive (DP), CD4+ or CD8+ single positive T cells were determined 24 h after a single injection or following a long-term treatment (twice a week for 4 weeks) with three different dilutions of ME which are corresponding to 4.5 ng/ kg, 22.5 ng/kg and 112.5 ng/kg doses of MLs. The apoptosis of the thymocytes was also tested by flow cytometry using Annexin V and propidium iodide. 24 h after a single injection of ME only the lowest dose caused in the blood samples an elevated CD4+/CD8+ ratio and in thymus an enhanced proliferation of DN thymocytes indicating a similar bell-shaped curve of immunological efficacy. After a treatment for four weeks these responses were less intensive indicating that none of the three doses are immunologically optimal. Surprisingly, both in the acute and in the long-term trial only the lower doses induced significant enhancements in the ratio of apoptotic thymocytes. In addition, ME inhibited the dexamethasone (DX)-induced reduction of DN cell count in thymus, as well as the DX-induced decrease of CD4+/CD8+ ratio and CD4+ cell level in peripheral blood. These in vivo results suggest that investigation of thymocytes in vivo can be helpful in the immunological dose-finding since standardized ME is able to modulate the proliferation and apoptosis of thymocytes with a bell-shaped curve of efficacy. In addition, ME may act lymphoprotectively during DX treatment.

Adjuvants, Immunologic↗
PubMed2006

In vitro immunoreactivity towards lectin-rich or viscotoxin-rich mistletoe (Viscum album L.) extracts Iscador applied to healthy individuals.

The aim of the study was to investigate how exposure to mistletoe extracts in vivo may influence cellular immune reactions by peripheral blood mononuclear cells (PBMC). 47 healthy individuals were exposed for twelve weeks either to Iscador Quercus special (IQ; rich in mistletoe lectin [ML]) (n = 16), to Iscador Pinus (IP; poor in ML but rich in viscotoxin) (n = 15), or to placebo (physiological saline) (n = 16). PBMC were isolated before exposure, during exposure in week 4, 8 and 12, as well as 13 weeks after last exposure (week 25) and incubated for 7 days with IQ or IP. Proliferative response and cytokine release were determined (type 1 cytokines: interferon [IFN]-gamma, tumor necrosis factor [TNF]-gamma, type 2 cytokines: interleukin [IL]-5, IL-13, macrophage/ monocyte-cytokines: IL-1, TNFalpha). IP- and IQ incubation of PBMC from non-exposed individuals lead to a significant increase of proliferation and production of all cytokines. There was a significant increase of IQ-induced TNFalpha (but not IL-1) production by PBMC in week 25 in individuals receiv-type 2-cytokines IL-5 and IL-13 significantly decreased during the observation period (p < 0.05 compared to placebo in week 8). These data indicate that IP seems to contain an antigen, which decreases type 2 reactivity. Furthermore, a component present preferentially in IQ seems to activate the production of the monocyte/ macrophage-related cytokine TNFalpha by PBMC. Intriguingly, also in individuals receiving placebo injections immunological alterations were detected during the observation period.

Adult↗
PubMed2006

Nitric oxide involvement in the anti-tumor effect of mistletoe (Viscum album L.) extracts Iscador on human macrophages.

Lectins from different types of mistletoe (Viscum album, VA) have cytotoxic and immunomodulatory properties that may be relevant in the inhibition of tumor growth. The mechanism of this anti-tumoral activity remains unknown, although recent investigations point out the induction of anti-tumoral cytotoxic T cell activation. In this study therapeutically available mistletoe extracts (Iscador) prepared from Quercus (VA-Q), apple (Malus, VA-M) or pine (Pinus, VA-P) were used to investigate their capacity to induce tumor regression through the modulation of another T helper-1 (Th-1)-mediated anti-tumoral activity: the activation of macrophages. Macrophages are essential targets for both pro- or anti-inflammatory drugs and constitute an essential member of the anti-tumoral immune response. Freshly isolated human monocyte-derived macrophages are activated and various VA extracts are directly incorporated to cultures to assay their properties on the inflammatory and/or tumor cytotoxic responses. The data indicate that immunomodulatory activities of VA extracts differ according to their origin. VA-M and VA-P were able to increase anti-tumoral activity of activated human macrophages, with a possible role for nitric oxide in this effect.

Antineoplastic Agents, Phytogenic↗
PubMed2006

Molecular mechanisms underlying the immunomodulatory effects of mistletoe (Viscum album L.) extracts Iscador.

Viscum album (VA) preparations (Iscador) consist of aqueous extracts from different types of European mistletoe. Biologically active components of VA extracts include mistletoe lectins (ML) and viscotoxins. The treatment with VA extracts or with purified ML has been shown to be associated with tumor regression in several in vivo experimental models of tumoral implantation. The mechanisms underlying the anti-tumoral activity of VA or ML are complex and involve apoptosis, angiogenesis and immunomodulation. This review provides an account of the current status of the understanding of the VA-associated immunomodulation in various cell types including lymphoblastoid, monocytic or endothelial cell lines.

Angiogenesis Inhibitors↗
PubMed2005

Safety and efficacy of the long-term adjuvant treatment of primary intermediate- to high-risk malignant melanoma (UICC/AJCC stage II and III) with a standardized fermented European mistletoe (Viscum album L.) extract. Results from a multicenter, comparative, epidemiological cohort study in Germany and Switzerland.

BACKGROUND: Mistletoe therapy is the most frequently used complementary treatment in cancer patients in Germany and Switzerland. However, its safety and efficacy were controversially discussed, also in case of malignant melanoma (MM). OBJECTIVES: The present study should evaluate the therapeutic safety and efficacy of a long-term therapy with a standardized fermented European mistletoe (Viscum album L.) extract Iscador (FME) during post-surgical aftercare of primary intermediate to high-risk MM (UICC/AJCC stage II-III) patients and compare it with an untreated parallel control group from the same cohort. METHODS: The study was designed as a multicenter, comparative, retrolective, epidemiological cohort study with parallel groups, carried out according to the guidelines of Good Epidemiological Practice (GEP). All patients suffered from surgically treated and histopathologically confirmed primary MM in UICC/AJCC stage II-III without distant metastases. In the study group, FME was administered subcutaneously 2-3 times weekly for at least three months, while the untreated control group was merely observed ("watchful waiting"). In both groups some patients also received radio-, chemo-, and/or immunotherapy. The patients were followed until the last visit or until death. Unselected, chronologically ordered, and standardized anonymous data from medical records that satisfied the predefined eligibility criteria were included for the "per protocol" analysis. Safety was assessed by the number of patients with FME-associated adverse drug reactions (ADRs) and by the search for tumor enhancement. The primary endpoint of efficacy was the adjusted tumor-related survival. Secondary end-points were the overall-, the disease-free- and the brain metastasis-free survival. The survival results were analyzed after adjustment for baseline imbalances, treatment regimens and other potential confounders by the Cox proportional hazard regression method. RESULTS: 686 eligible patients (329 FME vs. 357 controls) from 35 centers were observed for a median aftercare of 81 vs. 52 months. The median FME therapy duration was 30 months. At baseline, both groups were comparable concerning demography, tumor history and risk factors for progression. Additional adjuvant chemotherapy was more frequent in the study group, while immunotherapy was more frequent in the control group. Eleven patients (3.3 %) developed systemic ADRs attributed to the FME-treatment, and 42 patients (12.8 %) developed local ADRs, with mild to intermediate (WHO/CTC grade 1-2) ADR severity and spontaneous resolution in most cases. In six patients the ADRs resulted in therapy termination. Life-threatening ADRs, ADR-related mortality or tumor enhancement were not observed. On the contrary, the incidence rate of lung metastases and the adjusted hazard ratio for brain metastases were significantly lower in the FME group. In the course of the study and during aftercare a total of 212 (30.9 %) patients relapsed or progressed, and 107 (15.6 %) died. A significantly longer tumor-related survival was found in the FME group when compared with the untreated controls (unadjusted tumor-related mortality rate 8.9 % vs. 10.7 %, Kaplan-Meier estimate, Log-rank test, p = 0.017), which was confirmed after adjusting for potential confounders by the tumor-related mortality hazard ratio estimate HR (95 % confidence intervals) = 0.41 (0.23-0.71), p = 0.002. The adjusted HR results of the overall survival, disease-free survival, and the brain metastases-free survival were also significantly superior in the FME group. CONCLUSION: The long-term FME treatment in patients with primary intermediate to high-risk MM appears safe. Tumor enhancement was not observed. When compared with an untreated parallel control group from the same cohort, the results of the FME treatment suggested a significant survival benefit in primary stage II-III MM patients. These results on survival warrant reconfirmation in a prospective randomized clinical trial with optimized study design and treatment conditions.

Antineoplastic Agents, Phytogenic↗
PubMed1998

Subcutaneous infiltrates induced by injection of mistletoe extracts (Iscador).

Iscador, an aqueous extract of Viscum album L., has been widely used as an anti-cancer drug for several decades. Mistletoe lectins have the capacity to activate nonspecific defense mechanisms, and lectin-carbohydrate interactions may be involved in clinically applicable immunomodulation. During treatment with whole-plant mistletoe extract, an inflammatory reaction usually occurs at the site of the injection, early in therapy. These injection sites were examined histologically. Seven subjects received three subcutaneous injections of Iscador QuFrF or Iscador Qu Spezial (twice 0.1 mg and once 2.5 mg) during 9 days. In all subjects, examination of skin biopsies showed a normal epidermis. The dermal and subcutaneous regions contained a dense perivascular lymphocyte infiltrate and increased monocytes. We could not document any increase of plasma cells, eosinophils, mast cells, neutrophils, or granulocytes, as would be the case for a granulomatous infiltrate. In the blood, we observed a significant increase in neutrophils and monocytes 24 hours after administration of 2.5 mg of Iscador.

Adult↗