Article

Stimulation of natural killer cells by homoeopathic complexes: An in vitro and in vivo pilot study in advanced cancer patients

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Abstract

The present study was designed in order to evaluate the effects of five homoeopathic complex preparations on functional activity natural killer cells (NKCs) in advanced cancer patients. We examined the effects of Coenzyme Compositum®, Ubichinon Compositum®, Glyoxal Compositum®, Katalysatoren® and Traumeel® on the functional activity of NKCs. Experimental procedures included in vitro and in vivo trials. The in vitro trials were performed in NKCs isolated from 12 healthy volunteers (aged 44 ± 4 years) and incubated with the five homoeopathic complex preparations. The in vivo trials were performed in 15 advanced cancer patients (aged 55 ± 12 years) supplemented for 3 months with the homoeopathic preparations. All five homoeopathic preparations significantly increased the cytotoxic activity of the NKCs at the lowest NKCs/target cell ratio 12:1 (p < 0·05). The order of activity was: Ubichinon Compositum® > Glyoxal Compositum® > Katalysatoren® > Traumeel® > Coenzyme Compositum®. In the advanced cancer patients, the homoeopathic preparation significantly increased NKCs cytotoxic activity (p < 0·05). The homoeopathic complex preparations tested in this study can be used as an adjuvant immunotherapy in advanced cancer patients. Copyright © 2013 John Wiley & Sons, Ltd.

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... Concurrent immunotherapy uses immune cells to identify and destroy cancer and virally infected cells, reducing the tumor burden and potential for malignant transformation that conventional cancer treatment must address [28]. NKCs produce immunosurveillant cytokines IL-2, IL-12, IL-18, and IL-21. ...
... NKCs produce immunosurveillant cytokines IL-2, IL-12, IL-18, and IL-21. NKCs quickly and selectively kill cancer and virally infected cells via non major histocompatibility complex class-I (MHC-I) restricted action, before high levels of cell membrane MHC-I molecules are evident [28]. However, conventional chemotherapeutics such as bortezomib, chlorambucil, cladribine, docetaxel, MG-132, paclitaxel, and vinblastine, inhibit NKC-mediated cancer and virally infected cell killing, without preventing NKC activation [28]. ...
... NKCs quickly and selectively kill cancer and virally infected cells via non major histocompatibility complex class-I (MHC-I) restricted action, before high levels of cell membrane MHC-I molecules are evident [28]. However, conventional chemotherapeutics such as bortezomib, chlorambucil, cladribine, docetaxel, MG-132, paclitaxel, and vinblastine, inhibit NKC-mediated cancer and virally infected cell killing, without preventing NKC activation [28]. Thus, concurrent treatments able to activate NKCs complement conventional chemotherapeutics. ...
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Background: Homeopathy is used by 12 to 24% of European cancer patients, representing 40.4% of patients at European integrative cancer centers. In 2011, a Swiss literature review on homeopathy led to homeopathic treatment coverage in the Swiss national health insurance program. Homeopathy for curative pediatric cancer treatment is limited to 7.4% in the Netherlands, but, 76.5% of German parents will use homeopathy as part of their children’s cancer treatment. The purpose of this paper is to determine what is needed for homeopathy to play a larger role in curative, concurrent, and supportive cancer treatment. Methods: PubMed searches in September 2016 and January 2017 were performed with search terms “adverse effects, breast cancer, cancer, cervical cancer, endometrial cancer, homeopathy, ovarian cancer, prevention, treatment”. Curative, concurrent, and supportive homeopathic cancer treatments material was taken from these searches. Findings: At least five homeopathic formulations are immunologic adjuvants, activating natural killer cell destruction of cancer and virally infected cells. Ultramolecular Carcinosin, Phytolacca decandra, Conium, Thuja and Klimaktoplan® are appropriate for in vivo breast cancer trials. Lycopodium clavatum 5C and 15C are ready for in vivo cervical cancer trials. Sulphur 30C, may be considered for non-small cell lung adenocarcinoma treatment trials. Conventional cancer treatment associated anxiety, asthenia, depression, dermatitis, folliculitis, hot flushes, insomnia, nausea and vomiting, and stomatitis, respond to numerous homeopathic treatments including hetero-isotherapy. Conclusion & Significance: In vitro studies and retrospective case series indicate that homeopathy could provide curative cancer treatment for an array of cancers: Breast, cervix, gallbladder, liver, lung, oral, pancreas, periampullary, skin, and stomach. Appropriately designed randomized controlled trials (RCT) based on reproducible homeopathic treatments and clinical protocols, with intent-to-treat analysis will have increased validity. If these RCT have positive outcomes homeopathy will secure a position in curative, concurrent, and supportive cancer treatment.
... Concurrent immunotherapy uses immune cells to identify and destroy cancer and virally infected cells, reducing the tumor burden and potential for malignant transformation that conventional cancer treatment must address [28]. NKCs produce immunosurveillant cytokines IL-2, IL-12, IL-18, and IL-21. ...
... NKCs produce immunosurveillant cytokines IL-2, IL-12, IL-18, and IL-21. NKCs quickly and selectively kill cancer and virally infected cells via non major histocompatibility complex class-I (MHC-I) restricted action, before high levels of cell membrane MHC-I molecules are evident [28]. However, conventional chemotherapeutics such as bortezomib, chlorambucil, cladribine, docetaxel, MG-132, paclitaxel, and vinblastine, inhibit NKC-mediated cancer and virally infected cell killing, without preventing NKC activation [28]. ...
... NKCs quickly and selectively kill cancer and virally infected cells via non major histocompatibility complex class-I (MHC-I) restricted action, before high levels of cell membrane MHC-I molecules are evident [28]. However, conventional chemotherapeutics such as bortezomib, chlorambucil, cladribine, docetaxel, MG-132, paclitaxel, and vinblastine, inhibit NKC-mediated cancer and virally infected cell killing, without preventing NKC activation [28]. Thus, concurrent treatments able to activate NKCs complement conventional chemotherapeutics. ...
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Background: Conventional cancer treatment is associated with resistant cancer development, treatment and quality of life limiting adverse effects, and patients’ inability to complete intended treatment plans. Conventional cancer treatment’s adverse effects lead 36.1% of cancer patients to seek integrative cancer treatments, which can provide a 15 percentage-point improvement in cancer patients’ health status. Methods: PubMed searches in September 2016 and January 2017, and google scholar searches in August 2016 were performed with search terms “homeopathy or nutraceuticals or phytochemicals cancer treatment” from 2012 onwards. Adjuvant characteristics and adverse effects were taken from these searches. Supplemental specific hand searches were performed as needed. Findings: At least five homeopathic formulations are immunologic adjuvants, activating natural killer cell destruction of cancer and virally infected cells. Even delayed start homeopathy can extend survival. Organosulphurs are also immunologic chemosensitizers. Terpenes can inhibit or reverse drug resistance. Epigallocatechin modulates estrogen receptor expression. Polyunsaturated fatty acids are cancer cell membrane chemoradiation sensitizers. Esterified vitamin E analogues and Ayurvedic Triphala radiosensitize the MCF-7 breast cancer cell line. Curcumin and resveratrol also radiosensitize cancer cells. Withaferin A is synergistic with cisplatin permitting a smaller cisplatin dose while maintaining cisplatin’s effectiveness. Conventional cancer treatment associated anxiety, asthenia, depression, dermatitis, folliculitis, hot flushes, insomnia, nausea and vomiting, and stomatitis, respond to numerous homeopathic treatments including hetero-isotherapy. Conventional cancer treatment associated hand-foot syndrome, hematologic toxicity, mucositis, pain, sleep dysfunction, and overall toxicity respond to several nutraceuticals. Conclusion & Significance: The evidence for concurrent homeopathy and nutraceuticals use to prevent or limit conventional cancer treatment associated adverse effects exists. In vitro evidence suggests that homeopathy and nutraceuticals are effective chemoradiation sensitizers and adjuvants, with synergistic potential. Positive human trials would facilitate a broader and deeper role for homeopathy and nutraceuticals in integrative cancer treatment.
... Tr14 also consists of two mineral components (calcium sulfide and Mercurioamidonitrate). Evidence for immunomodulatory effects of Tr14 comes from humans [19][20][21], animals [22,23], and in vitro [21,24] studies. Clinical evidence for Tr14 and composition in more detail was reviewed by Schneider [25]. ...
... Tr14 also consists of two mineral components (calcium sulfide and Mercurioamidonitrate). Evidence for immunomodulatory effects of Tr14 comes from humans [19][20][21], animals [22,23], and in vitro [21,24] studies. Clinical evidence for Tr14 and composition in more detail was reviewed by Schneider [25]. ...
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The present double-blind, randomized, placebo-controlled clinical trial intended to test whether ingestion of a natural combination medicine (Tr14 tablets) affects serum muscle damage and inflammatory immune response after downhill running. 96 male subjects received Tr14 tablets, which consist of 14 diluted biological and mineral components, or a placebo for 72 h after the exercise test, respectively. Changes in postexercise levels of various serum muscle damage and immunological markers were investigated. The area under the curve with respect to the increase ( A U C i ) of perceived pain score and creatine kinase (CK) were defined as primary outcome measures. While for CK the p value of the difference between the two groups is borderline, the pain score and muscle strength were not statistically significant. However, a trend towards lower levels of muscle damage (CK, p = 0.05 ; LDH, p = 0.06 ) in the Tr14 group was shown. Less pronounced lymphopenia ( p = 0.02 ), a trend towards a lower expression of CD69 count ( p = 0.07 ), and antigen-stimulated ICAM-1 ( p = 0.01 ) were found in the verum group. The Tr14 group showed a tendentially lower increase of neutrophils ( p = 0.10 ), BDNF ( p = 0.03 ), stem cell factor ( p = 0.09 ), and GM-CSF ( p = 0.09 ) to higher levels. The results of the current study indicate that Tr14 seems to limit exercise-induced muscle damage most likely via attenuation of both innate and adaptive immune responses. This study was registered with ClinicalTrials.gov ( NCT01912469 ).
... The homeopathic preparations significantly increased the toxic activity of NKC cells (p <0.05). The combination homeopathic products tested in this study can be used as adjunctive therapy, immunotherapy in very advanced cancer [16]. ...
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IN VITRO EFFECTS OF HOMEOPATHIC DRUGS Christer Sundqvist Petrafoundation, Helsinki, Finland https://petrafoundation.com/en/blog/scientific-evidence-for-homeopathy-part-4 Published: 05.30.2020 The well-known and respected Finnish homeopath Jouni Jämsä (http://www.jounijamsa.fi/ ) continues his homeopathy research with us. In the first part, we talked to him about homeopathy in general and diluted homeopathic preparations in particular [1]. In the next section we investigated how plants respond to homeopathic preparations [2]. In the third part, we went through studies where homeopathy has been used for cancer patients [3]. In this fourth section, we investigate the effect of homeopathic preparations at the cellular level. Highly diluted homeopathic preparations are showing measurable effects in cell cultures [4]. Several studies have found that homeopathic agents give rise to apoptosis (programmed cell death). This phenomenon is a normal part of individual tissue development and regeneration. It is controlled by genes that regulate cell division [5, 6]. Apoptosis plays an important role in fetal development, regulation of the immune response, elimination of infected and transformed cells, and regulation of tissue size. Excessive apoptosis can lead to developmental disorders and degenerative diseases, while its absence can lead to autoimmune diseases, long-term viral infections and cancer. Utilization of drug apoptosis is already routine in the treatment of cancer. Drugs for the treatment of degenerative diseases that prevent apoptosis are likely to emerge in the market in the near future [7]. Apoptosis is something you want to achieve when treating cancer. Because cancer is a disease that affects many people and is very serious, researchers have been interested in different types of treatment options. The effect of homeopathy in cancer can also be studied with cell models. The potent homeopathic drug Lycopodium clavatum (5C and 15C) has anti-cancer activity on HeLa cells in vitro. This has been studied in the Laboratory of Cell Genetics and Molecular Biology at the University of Kalyan, India. The purpose of this study was to evaluate whether homeopathic highly diluted and potent preparations of Lycopodium Clavatum (LC-5C and LC-15C) have anti-cancer effects on HeLa cells. Cells were exposed to either LC-5C (diluted below Avogadro's number) or diluted above Avogadro's number (LC-15C). The results revealed that administration of Lycopodium had little or no toxic effects on the cells in the bloodstream, but caused marked apoptosis in cancer cells (HeLa), which appeared in the form of initial degradation of DNA. Highly diluted, dynamic homeopathic drugs, both below and above the Avogadro's number, caused cell death in cancer cells, suggesting that these drugs could potentially be used as supportive cancer care [8]. Frenkel's team at the University of Texas (M.D. Anderson Cancer Center, Texas, USA) performed cell experiments with highly diluted Indian homeopathic preparations (Carcinos, Phytolacca, Conium and Thuja). The preparations were tested with different breast cancer cell lines. The preparations were toxic to the cells, leading to cell cycle breakdown and cell death. Thus, these natural, diluted preparations had biological effects that should be further investigated [9]. The same was also supported by Psorinum 6 ×, which triggers apoptosis signals in human lung cancer cells. Studies of the effects of homeopathic Psorinum 6x on cell survival were initially performed in several cancer cell lines, including A549 (lung cancer cell line), HepG2 (liver cancer cell line) and MCF-7 (breast cancer cell line). The experiment investigated the therapeutic effects on cell cycle breakdown, cell death, reactive oxygen radical formation (ROS), and changes in mitochondrial membrane potential (MMP) using flow cytometry and fluorescence microscopy. It was found that treatment of cancer cell lines with Psorinum resulted in increased anti-cancer effects in A549 cells (lung cancer cell line) to a greater extent than in others. Psorinum prevented cell division after 24 hours of treatment and retained the cells in the G1 phase. It also caused e.g. ROS formation, MMP depolarization, morphological changes and DNA damage. The researchers concluded that Psorinum 6 × triggered apoptosis in A549 cells (lung cancer cells) via signaling proteins [10]. Condurango 6C and 30C also trigger apoptosis in lung cancer cells. The more diluted preparation was more effective [11]. The effects of homeopathy on programmed cell death have been extensively studied. Further support for this effect on apoptosis, which is advantageous in the treatment of cancer, has been obtained in e.g. Shagun Arora laboratory at Jaypee University of India. It was found that undiluted and diluted homeopathic preparations were toxic to cultured cancer cells. Homeopathic Sarsaparilla preparations were tested in isolated renal adenocarcinoma cell cultures, the Ruta graveolens preparation in cultured colon cancer cells and the Phytolacca decandra preparation in breast cancer cells. The results showed that all of the homeopathic preparations showed toxic effects in said cell cultures. The undiluted preparations had the best effect, but diluted preparations also served as a starting point of apoptosis. Homeopathic preparations have been shown to be anti-cancer drugs and further research can be encouraged [12]. The homeopathic preparation Calcarea carbonica caused apoptosis in cell cultures from cancerous mice. The study showed evidence of the so-called immunomodulatory mechanism of cell death [13]. In leukemia, homeopathic dilutions of Amanita phalloides have been successfully tested in cell cultures [14]. There is no placebo effect in cultured cancer cells, so here we have evidence that homeopathy is not just a placebo. Should homeopathic significance be increased in established medicine, more evidence from well-controlled and high-quality studies is needed [15].
... These homeopathic agents significantly increased the number of natural killer cells (NK). The study summarizes the suitability of these homeopathic drugs for the treatment of immunity in cancer patients [13]. ...
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Homeopathy utilizes the body's own resources and homeopathic preparations, for example, strengthen the immune system. A strong immune system plays a very important role in the treatment of cancer. Therefore, homeopathy is widely used in the treatment of cancer and there is plenty of clinical research evidence. Cancer is treated differently in established medicine compared to homeopathic methods. Both methods are needed to complement each other. For example, after cancer surgery and radiotherapy, the treatment can preferably continue utilizing homeopathic methods, since cytostatic drugs can lead to a patient poorly recovering from cancer surgery or cancer radiation due to an immune system not working properly. Homeopathic methods offer a gentle and effective solution for those who are recovering from cancer.
... The kit used for the evaluation of NKC cytotoxicity was "NKTEST" of ORPEGEN Pharma, Germany. The estimation of cytotoxicity of NKCs was performed by flow cytometry (Toliopoulos et al., 2013). The same measurements were repeated after the incubation of cells (for 150 minutes) with 10 mg/ml of the V. album L. + A. Alba extract. ...
Article
Viscum album L. is a widely used medicinal plant in cancer treatment, known since ancient times. The aim of this study was to investigate the antimetastatic properties of the ethanolic extracts of synergistic plants V. album L. (epiphyte) and Abies alba (host) used alone or in combination (mixture). Inhibition of platelet aggregation was evaluated in washed rabbit and human platelets. Levels of thromboxane A2 (TXA 2) were estimated by radioimmune assay and natural killer cells (NKCs) cytotoxicity by flow cytometry. The antimetastatic properties of V. album L. and A. alba were studied in a tumor-bearing Wistar rat model. All extracts inhibited platelet aggregation in a dose-dependent manner as well as TXA 2 production by three pathways of aggregation (adenosine diphosphate, platelet-activating factor, and arachidonic acid) while the mixture significantly increased NKCs cytotoxicity against cancer cells. In tumor-bearing Wistar rats, the treatment with the mixture of V. album L. and A. alba extract reduced the metastatic locations almost by 77%. These data suggest that V. album L. and A. alba extract reduced metastasis through inhibition of platelet aggregation and amplification of the body defense mechanisms against cancer cells.
Thesis
Hintergrund und Ziele: Das Ziel Integrativer Medizin (IMed) in der Onkologie ist es, PatientInnen in ihrem Krankheitsverlauf zu begleiten, Nebenwirkungen der Erkrankung selbst sowie der notwendigen Karzinomtherapien zu mildern und die Lebensqualität zu verbessern. Da Karzinomerkrankungen häufig mit Vitaminmangelzuständen bei den Betroffenen einhergehen, ist die Substitution von Vitaminen Bestandteil integrativmedizinischer Maßnahmen. In der Integrativen Sprechstunde der Universitätsfrauenklinik Erlangen werden KarzinompatientInnen deshalb fünf verschiedene IMed-Vitaminfusionen für unterschiedliche Indikationen angeboten, deren Mikronährstoffrezepturen zusammen mit der Klinikapotheke entwickelt wurden. Die vorliegende Arbeit beschäftigt sich mit der subjektiven Bewertung der intravenösen Vitamintherapie und der Zufriedenheit der Patientinnen in Bezug auf die verschiedenen Aspekte der IMed-Infusionstherapie. Methoden: Das untersuchte Patientinnenkollektiv dieser retrospektiven Beobachtungsstudie umfasst 59 Patientinnen des Universitätsklinikums Erlangen mit Mammakarzinom oder anderen gynäkologischen Tumorerkrankungen. Ihre Zufriedenheit mit den IMed-Infusionen und ihre subjektive Beurteilung wurde mittels eines validierten und standardisierten Fragebogens erhoben. Ergebnisse und Beobachtungen: Das befragte Kollektiv bestand zu 79,7 % aus Patientinnen mit Mammakarzinom und zu 20,3 % aus Patientinnen mit gynäkologischen Tumoren. Insgesamt fanden 351 Infusionen statt. Davon macht die IMed-Regenerationsinfusion mit insgesamt 216 Anwendungen den Hauptteil aus (61,5 %), gefolgt von der IMed-Immuninfusion (27,6 %). Mit den organisatorischen Rahmenbedingungen der Infusionen zeigten sich die Patientinnen überwiegend zufrieden. 62,7 % konnten eine Verbesserung der Lebensqualität feststellen und 52,5 % gaben Beschwerdelinderungen an. Am häufigsten konnten dabei Müdigkeit, Abgeschlagenheit und Antriebslosigkeit gemildert werden. Insgesamt zeigt sich eine hohe Zufriedenheit mit der Verträglichkeit und eine geringe Nebenwirkungsrate. Patientinnen, die eine Linderung der Beschwerden angaben, unterschieden sich von Patientinnen ohne Beschwerdelinderung durch eine höhere Anzahl an Infusionen, sowie ihren Therapiestatus. So berichteten Patientinnen der Nachsorge seltener über eine Beschwerdelinderung als Patientinnen, die neoadjuvant, adjuvant oder palliativ mit systemischen Therapien behandelt wurden. Eine laufende Therapie mit den IMed-Infusionen ging ebenfalls häufiger mit der Angabe von Beschwerdelinderungen einher. Der Großteil der Patientinnen (78,0 %) würde die IMed-Infusionen weiterempfehlen. Die Weiterempfehlung hing von der Einschätzung der Verträglichkeit und Wirksamkeit, sowie vom Infusionsstand (abgeschlossene/ andauernde IMed-Infusionstherapie) ab. Patientinnen unter laufender Infusionstherapie hatten eine höhere Weiterempfehlungsrate. Schlussfolgerung: Die Ergebnisse lassen eine positive Wahrnehmung und Eignung des Therapieangebotes mit Mikronährstoffinfusionen erkennen. Unter Berücksichtigung von Kontraindikationen, Nebenwirkungen und möglichem Nutzen können die Infusionen KarzinompatientInnen u.a. durch Linderung von Symptomen, Verbesserung der Lebensqualität und Stärkung der Selbstwirksamkeit in ihrem Krankheitsverlauf unterstützen. Standardisierungsprozesse, wie in der Integrativen Sprechstunde durchgeführt, sowie die Evaluation integrativmedizinischer Angebote gewährleisten eine patientenzentrierte Betreuung nach aktuellem wissenschaftlichem Standard
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This study was conducted to evaluate the effect of two homeopathic complexes Ubichinon compositum® (Ubi comp) and Coenzyme compositum ad us. vet.® (CoQ10 comp) on bovine sperm mitochondrial activity. Sperm viability, acrosomal integrity and sperm chromatin structure were estimated to detect the possible side effect of complexes on other sperm parameters. Mitochondrial activity was significantly enhanced by both Ubi comp (P<0.01) and CoQ10 comp (P<0.05). No effects were detected in other tested sperm parameters. The tested homeopathic complex medicines stimulate the mitochondrial activity of bovine sperm without effects on their viability, acrosomal integrity or chromatin structure. The possibility that this translates into improved fertilization capacity in artificial insemination should investigated.
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Ratios of urinary 8-hydroxy-2′-deoxyguanosine to urinary creatinine (8-OHdG/creatinine) have been considered as a good biological indicator of DNA oxidation. Urinary 8-OHdG/creatinine levels of lung cancer patients were evaluated by enzyme-linked immunosorbent assay using a monoclonal antibody N45.1 during radiotherapy and chemotherapy. An increase in urinary 8-OHdG/creatinine was found in non-small-cell carcinoma (non-SCC) patients during the course of radiotherapy. SCC patients showed higher levels of urinary 8-OHdG/creatinine than the controls. Furthermore, SCC patients with complete or partial response to the chemotherapy showed a significant decrease in urinary 8-OHdG/creatinine while patients with no change or progressive disease showed an increase.
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That the inheritance of mutations in tumor susceptibility genes alone cannot determine risk for developing cancer is now well accepted. Immune functions have long been recognized as one of the important risk modifying factors in this regard. In an attempt to develop a multiparametric approach to identify high risk individuals from cancer families, we have examined NK cell function in unaffected members from familial breast cancer families. We have also carried out a parallel study of T lymphocyte functions in these individuals. Our studies demonstrate a significantly lower NK cell activity in members from cancer families. T lymphocyte activity also showed a similar trend, with the unaffected members demonstrating a notably lowered T lymphocyte function. In addition the data from patients reveals differential sensitivity of NK and T lymphocyte function to the disease phenotype. Implications of these observations are discussed.
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Asparagus racemosus Linn. (Fam. Liliaceae) is an ethno-pharmacologically acclaimed Ayurvedic medicinal plant. In the present study, aqueous extract of A. racemosus (ARC) was fractionated and screened for the polysaccharide fraction (ARP). The characterization was done by enzymatic, Size Exclusion, gas chromatography with flame ionization detector (GC-FID), high pressure anion exchange chromatography (HPAEC) and thin layer chromatographic analyses. Phyto-chemical evaluation confirmed the presence of 26.7% of 2→1 linked fructo-oligosaccharides (FOS). They have a degree of polymerization (DP) of nearly 7-8. Cytotoxicity evaluation on P388 cell lines was consistent with low cytotoxicity of the extracts. In vitro Natural Killer (NK) cell activity was evaluated using human peripheral blood mononuclear cells (PBMC) isolated from whole blood on a ficoll-hypaque density gradient. K562 a myeloid leukemia cell line, were used as target cells. ARC, tested over the range 0.2-50 μg/ml, showed a dose-related stimulation of NK cell activity with a peak increase of 16.9±4.4% at 5.6 μg/ml. However, ARP demonstrated a higher stimulatory activity of 51.8±1.2% at 25 μg/ml. The results indicate that the FOS from A. racemosus potentiates the NK cell activity and this could be an important mechanism underpinning the 'Rasayana' properties of this plant.
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As part of the innate immune response NK cells destroy infected, transformed, or otherwise stressed cells within hours of activation. In contrast, CD4(+) T lymphocytes require a sustained increase in their metabolism in order to cope with the biogenesis of cell components, in a process of proliferation and differentiation into effector cells. Recently, mitochondria have been implied in T lymphocyte immune synapse function but little is known on the role of mitochondria in the NK cell interaction with tumour cells. Here we analysed NK cells mitochondrial membrane potential (Deltapsi(m)) as an indicator of mitochondrial energy status and cellular homeostasis. Upon contact with K562 tumour cells, NK cells undergo Deltapsi(m) depolarization, indicating a rapid consumption of their metabolic energy. Furthermore, pharmacological inhibition of ATP synthesis down-regulates NK cell cytotoxic activity. Confocal- and electron-microscopy analyses showed re-organization of NK cells mitochondria towards the site of interaction with K562 tumour cell (NK cell immune synapse), perhaps as a way to compensate for local energy consumption. Interestingly, mitochondrial re-organization also takes place following NK stimulation with anti-NKGD2 antibodies but not with anti-KIR2DL1 antibodies, suggesting that activating rather than inhibiting cell signalling, triggered by NK cell receptors, is involved in NK cell mitochondria dynamics.
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This study addresses the interleukin (IL)-21 effects on resting peripheral blood natural killer (NK) cells in chronically HIV-infected individuals. The effects of IL-21 on perforin expression, proliferation, degranulation, interferon (IFN)-gamma production, cytotoxicity and induction of STAT phosphorylation in NK cells were determined in vitro. Peripheral blood mononuclear cells from HIV-infected and healthy individuals were incubated in vitro for 6 h, 24 h or 5 days with IL-21 or IL-15. Percentages of perforin, IFN-gamma, CD107a, NKG2D and STAT3-5 positive cells were determined within NK cell populations. K562 cells were used as target cells in NK cytotoxicity assay. Frequency of CD56 cells in chronically HIV-infected individuals was diminished. Perforin expression in CD56 and CD56 was comparable in healthy and HIV-infected individuals. IL-15 upregulated perforin expression primarily in CD56 NK cells, whereas IL-21 upregulated perforin in both NK subsets. IL-21 and IL-15 upregulated CD107a and IFN-gamma, as well as NK cytotoxicity. IL-15 predominantly activated STAT5, whereas IL-21 activated STAT5 and STAT3. IL-15, but not IL-21 increased NK cell proliferation in uninfected and HIV-infected individuals. IL-21 augments NK effector functions in chronically HIV-infected individuals and due to its perforin enhancing properties it has potential for immunotherapy or as a vaccine adjuvant.
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The present study was designed to evaluate the chemotherapy-induced cellular immunosuppression in 20 children with acute lymphoblastic leukemia (ALL) in remission and receiving maintenance chemotherapy. Peripheral blood was serially obtained from leukemic children during vincristine/cyclophosphamide/6-mercaptopurine/prednisone combined consolidation chemotherapy. The mean absolute number of peripheral blood lymphocytes as well as the mean absolute numbers of lymphocyte subsets (T cells, T cell subsets, B cells, and natural killer cells) from leukemic children before consolidation chemotherapy were all significantly lower than in control subjects; however, the percentages of lymphocyte subsets were similar in both groups. After consolidation chemotherapy, the percentages of CD4+ T lymphocytes and natural killer (NK) cells were significantly decreased and the percentages of monocytes and CD8+ T lymphocytes were significantly increased. Phytohemagglutinin- and 12-O-tetradecanoylphorbol-13-acetate-induced production of interleukin-2 (IL-2) and NK-cell-mediated cytotoxic activity by peripheral blood mononuclear cells (PBMC) were also substantially decreased in the post-therapy groups. NK activity correlated with the percentage of NK cells in PBMC. In contrast, OK432-induced production of tumor necrosis factor alpha (TNF alpha) and killer activity against NK-resistant target cells were significantly increased after therapy as compared with the pre-therapy and control groups. TNF alpha production correlated with the percentage of monocytes in PBMC. These results demonstrate that substantial quantitative and qualitative chemotherapy-induced abnormalities of the cellular immune system are present in the majority of patients treated with ALL. It is also suggested that the increased TNF alpha production by monocytes and the appearance of potent killing activity against NK-resistant targets might compensate for the defects of IL-2 production and NK activity during intensive consolidation chemotherapy.
Article
The direct effect of selected cytostatic drugs on natural killer (NK) cell activity was evaluated. Peripheral blood mononuclear cells from healthy donors were tested for their cytolytic activity in vitro in the presence of adriamycin, methotrexate, leucovorin, vincristin, cytosine arabinoside and teniposide. Most of the tested cytostatic drugs did not show to be active at concentrations comparable to their plasma level. However, diluents of some preparations (cytosine arabinoside, teniposide) containing organic solutions and stabilizing additives (e.g. benzyl alcohol) suppressed the NK activity more than chemotherapeutic agents alone. Thus teniposide, containing such additives, inhibited NK activity already at 5 mg/ml, while its peak plasma concentration was 23.8 mg/ml. The inhibitory concentrations of teniposide did not affect the target binding of effector cells and the expression of 14 tested leukocyte differentiation markers. This implies that a postbinding step of the lytic process was altered by the preparation. Likewise, no inhibition of lectin dependent cellular cytotoxicity by teniposide and its diluent was observed, suggesting that the lectin may substitute the missing lytic signal.
Article
Studies of cytotoxicity by human lymphocytes revealed not only that both allogeneic and syngeneic tumor cells were lysed in a non-MHC-restricted fashion, but also that lymphocytes from normal donors were often cytotoxic. Lymphocytes from any healthy donor, as well as peripheral blood and spleen lymphocytes from several experimental animals, in the absence of known or deliberate sensitization, were found to be spontaneously cytotoxic in vitro for some normal fresh cells, most cultured cell lines, immature hematopoietic cells, and tumor cells. This type of nonadaptive, non-MHC-restricted cellmediated cytotoxicity was defined as “natural” cytotoxicity, and the effector cells mediating natural cytotoxicity were functionally defined as natural killer (NK) cells. The existence of NK cells has prompted a reinterpretation of both the studies of specific cytotoxicity against spontaneous human tumors and the theory of immune surveillance, at least in its most restrictive interpretation. Unlike cytotoxic T cells, NK cells cannot be demonstrated to have clonally distributed specificity, restriction for MHC products at the target cell surface, or immunological memory. NK cells cannot yet be formally assigned to a single lineage based on the definitive identification of a stem cell, a distinct anatomical location of maturation, or unique genotypic rearrangements.
Article
Natural killer (NK) cells, the CD3- CD56+ CD16+ subset of peripheral blood lymphocytes, have long been known to be involved in non-major histocompatibility complex-restricted natural immunity to virally infected and malignant target cells. The association of abnormalities in NK cell numbers or functions with a broad spectrum of human diseases has been more clearly defined in recent years as a result of the improved knowledge of NK cell physiology and advances in monitoring of NK cell functions in health and disease. The ability to reliably measure changes in NK activity and/or numbers during the course of disease or response to treatment has focused attention on the role of the NK cell in disease pathogenesis. The improved understanding of NK cell deficiency in disease has opened a way for therapies specifically designed to improve NK cell function. The therapeutic use of biologic response modifiers capable of augmenting NK cell activity in vivo and of adoptive transfer of highly enriched, activated autologous NK cells in diseases such as cancer and AIDS is being evaluated. The importance of NK cells in health and the consequences of NK cell deficiency or excess are likely to be more extensively monitored in the future.
Article
Fas ligand (FasL) on cytotoxic lymphocytes is important for mediating apoptosis of activated lymphocytes and other target cells. We have reported that NK cell functions, such as proliferation, cell death, and killing activity, are subject to regulation by cellular redox status. Here, we report that expression of FasL protein and mRNA in activated NK cells is also regulated by redox. Ligation of CD16 on IL-2-preactivated NK cells resulted in reduction of intracellular peroxide level as well as induction of FasL expression. This CD16-induced FasL expression was suppressed by oxidative stress, including thiol deprivation or treatment with hydrogen peroxide (H2O2). Addition of thiol-reducing compounds, such as L-cystine, 2-ME, or N-acetyl cysteine, restored FasL expression. These data suggest that CD16 stimulation requires cellular reducing status for FasL induction in NK cells. Because FasL gene activation following CD16 cross-linking is regulated by the NF of activated T cells (NFAT), we examined the effect of oxidative stresses on NFAT activation. Electrophoretic mobility shift assays revealed that both thiol insufficiency and H2O2 treatment suppressed DNA-binding activity of NFAT and that addition of thiol-reducing compounds reversed or even enhanced it. Furthermore, these oxidative stresses inhibited activity of calcineurin, a serine/threonine phosphatase that regulates NFAT activation. These results suggest that suppression of calcineurin and NFAT activation is a mechanism by which oxidative stress inhibits FasL induction in activated NK cells and further support the hypothesis that thiol-reducing compounds might be required for maintenance of optimal NK functions under physiologic oxidative conditions.
Article
The humanized monoclonal antibody Herceptin, which specifically targets HER-2/neu, exhibits growth inhibitory activity against HER-2/neu-overexpressing tumors and is approved for therapeutic use with proved survival benefit in patients with HER-2/neu-positive breast cancer. In the present study, we investigated whether Herceptin could affect the HER-2/neu-overexpressing gastric cancer cells based on antibody-dependent cell-mediated cytotoxicity (ADCC) and compared immune effector cells from gastric cancer patients with normal individuals on ADCC. HER-2/neu-expressing gastric cancer cells could be killed by Herceptin-mediated ADCC and the Herceptin-induced ADCC correlated with the degree of HER-2/neu expression on the gastric cancer cells. However, the Herceptin-mediated ADCC was significantly impaired in peripheral blood mononuclear cells from advanced disease patients (n = 10) compared with that in early disease (n = 12; P = 0.04) or healthy individuals (n = 10, P = 0.02). Moreover, natural killer (NK) cells purified from patients with advanced disease indicated less Herceptin-mediated ADCC in comparison with that from healthy donors (P = 0.04), whereas monocytes purified from the patients showed an almost equal amount of Herceptin-mediated ADCC in comparison with that from healthy individuals, indicating that NK cell dysfunction contributed to the impaired Herceptin-mediated ADCC in gastric cancer patients. Furthermore, the NK-cell dysfunction on Herceptin-mediated ADCC correlated with the down-regulation of CD16zeta expression in the patients, and interleukin 2 ex vivo treatment of NK cells could restore the impairment of Herceptin-mediated ADCC, concomitant to the normalization of the expression of CD16zeta molecules. Thus, some modalities such as interleukin 2 treatment aimed at reversing NK dysfunction may be necessary for successful Herceptin treatment of gastric cancer.
Article
Natural killer (NK) cells are directly cytotoxic for tumor cells and play a primary role in regulating immune responses. We monitored levels of NK cell cytotoxic activity in cancer patients receiving D-Fraction extracted from maitake mushrooms (Grifola frondosa). Elevated levels of cytotoxic activity were maintained for one year. To elucidate the mechanisms underlying long-term activation of NK cells during treatment with D-Fraction, we examined tumor volume and levels of IFN-gamma and TNF-alpha in MM46-bearing C3H/HeN mice to which D-Fraction was administered for 19 d. D-Fraction markedly suppressed tumor growth, corresponding with increases in TNF-alpha and IFN-gamma released from spleen cells and a significant increase in TNF-alpha expressed in NK cells. This suggests that the D-Fraction activates NK cells even on the 20th day after treatment. Furthermore, D-Fraction increased macrophage-derived interleukin (IL)-12, which serves to activate NK cells. These results suggest that NK cells are not only responsible for the early effects of D-Fraction on tumor growth, but also for the long-term tumor-suppressive effects of D-Fraction through increased IL-12 released from macrophages.
Article
There are a surprisingly large number of human natural killer (NK) cell deficiency states that provide insight into the role of NK cells in defense against human infectious disease. Many disorders associated with NK cell defects are caused by single gene mutations and, thus, give additional understanding concerning the function of specific molecules in NK cell development and activities. A resounding theme of NK cell deficiencies is susceptibility to herpesviruses, suggesting that unexplained severe herpesviral infection should raise the possibility of an NK cell deficit.
Article
Aged garlic extract (AGE) has manifold biological activities including immunomodulative and antioxidative effects. It is used as a major component of nonprescription tonics and cold-prevention medicines or dietary supplements. Advanced-cancer patients decline in immune functions and quality of life (QOL). The study's subjects were patients with inoperable colorectal, liver, or pancreatic cancer. In a randomized double-blind trial, AGE was administered to one group and a placebo was administered to another for 6 mo. The primary endpoint was a QOL questionnaire based on the Functional Assessment of Cancer Therapy (FACT). The subendpoints were changes in the natural-killer (NK) cell activity the salivary cortisol level from before and after administering AGE. Out of 55 patients invited to participate in the trial, 50 (91%) consented to enroll. They consisted of 42 patients with liver cancer (84%), 7 patients with pancreatic cancer (14%), and 1 patient with colon cancer (2%). Drug compliance was relatively good in both the AGE and placebo groups. Although no difference was observed in QOL, both the number of NK cells and the NK cell activity increased significantly in the AGE group. No adverse effect was observed in either group. The study showed that administering AGE to patients with advanced cancer of the digestive system improved NK cell activity, but caused no improvement in QOL.
Article
Much debate has arisen about whether antioxidant supplementation alters the efficacy of cancer chemotherapy. Some have argued that antioxidants scavenge the reactive oxygen species integral to the activity of certain chemotherapy drugs, thereby diminishing treatment efficacy. Others suggest antioxidants may mitigate toxicity and thus allow for uninterrupted treatment schedules and a reduced need for lowering chemotherapy doses. The objective of this study is to systematically review the literature in order to compile results from randomized trials that evaluate concurrent use of antioxidants with chemotherapy. MEDLINE, Cochrane, CinAhl, AMED, AltHealthWatch and EMBASE databases were searched. Only randomized, controlled clinical trials that reported survival and/or tumor response were included in the final tally. The literature searches were performed in duplicate following a standardized protocol. No meta-analysis was performed due to heterogeneity of tumor types and treatment protocols used in trials that met the inclusion criteria. Of 845 articles considered, 19 trials met the inclusion criteria. Antioxidants evaluated were: glutathione (7), melatonin (4), vitamin A (2), an antioxidant mixture (2), vitamin C (1), N-acetylcysteine (1), vitamin E (1) and ellagic acid (1). Subjects of most studies had advanced or relapsed disease. None of the trials reported evidence of significant decreases in efficacy from antioxidant supplementation during chemotherapy. Many of the studies indicated that antioxidant supplementation resulted in either increased survival times, increased tumor responses, or both, as well as fewer toxicities than controls; however, lack of adequate statistical power was a consistent limitation. Large, well-designed studies of antioxidant supplementation concurrent with chemotherapy are warranted.
Article
Cancer immune surveillance is considered to be an important host protection process to inhibit carcinogenesis and to maintain cellular homeostasis. In the interaction of host and tumour cells, three essential phases have been proposed: elimination, equilibrium and escape, which are designated the 'three E's'. Several immune effector cells and secreted cytokines play a critical role in pursuing each process. Nascent transformed cells can initially be eliminated by an innate immune response such as by natural killer cells. During tumour progression, even though an adaptive immune response can be provoked by antigen-specific T cells, immune selection produces tumour cell variants that lose major histocompatibility complex class I and II antigens and decreases amounts of tumour antigens in the equilibrium phase. Furthermore, tumour-derived soluble factors facilitate the escape from immune attack, allowing progression and metastasis. In this review, the central roles of effector cells and cytokines in tumour immunity, and the escape mechanisms of tumour cells during tumour progression are discussed.
Article
Natural killer (NK) cells are effector lymphocytes of the innate immune system that control several types of tumors and microbial infections by limiting their spread and subsequent tissue damage. Recent research highlights the fact that NK cells are also regulatory cells engaged in reciprocal interactions with dendritic cells, macrophages, T cells and endothelial cells. NK cells can thus limit or exacerbate immune responses. Although NK cells might appear to be redundant in several conditions of immune challenge in humans, NK cell manipulation seems to hold promise in efforts to improve hematopoietic and solid organ transplantation, promote antitumor immunotherapy and control inflammatory and autoimmune disorders.
Article
Natural killer (NK) cells circulate through the blood, lymphatics and tissues, on patrol for the presence of transformed or pathogen-infected cells. As almost all NK cell receptors bind to host-encoded ligands, signals are constantly being transmitted into NK cells, whether they interact with normal or abnormal cells. The sophisticated repertoire of activating and inhibitory receptors that has evolved to regulate NK cell activity ensures that NK cells protect hosts against pathogens, yet prevents deleterious NK cell-driven autoimmune responses. Here I highlight recent advances in our understanding of the structural properties and signaling pathways of the inhibitory and activating NK cell receptors, with a particular focus on the ITAM-dependent activating receptors, the NKG2D-DAP10 receptor complexes and the CD244 receptor system.
Article
The expression of NK cells activation receptors was assessed by comparative study of two groups of women workers at a chemical reagents factory, located in Zapopan, Jalisco, Mexico. Twenty of them were exposed to environmental toxics identified and quantified by gas chromatography, and 20 women unexposed to toxic substances. The expression of the surface markers CD56+ and CD3+, and of the activation receptors and co-receptors on NK cells was quantified by flow cytometry. To assess the cellular damage produced by chronic exposure to the toxics, the thiobarbituric acid reacting substances (TBARS) generated and the total plasma antioxidizing capacity (TPAC) were quantified in both groups. The exposed women had been exposed at least to 12 volatile toxic compounds, benzene, benz(a)pyrene, ethylbenzene, dimethylbenz(a)anthracene, xylene, toluene, styrene, chloroform, formaldehyde, iodine, chlorine and fluorine. Significant difference between the two groups was in the proportion of CD3 lymphocytes, 72.7+/-10.3% in the unexposed women versus 66.8+/-7.9% in the exposed group (p<0.05). The density of expression of NKG2D and NKp30 receptors was significantly higher in the unexposed women compared to the exposed group: NKG2D were 31.3+/-6.3 and NKp30 were 9.5+/-5.2 in the unexposed women and 5.14+/-2.9 (p<0.01) and 4.6+/-1.9 (p<0.05), respectively in the exposed women. No statistically significant differences were found in the expression of NKp80, NKp46 and 2B4 receptors. The concentration of TBARS was lower in women from the unexposed group than the corresponding data from women of the exposed group. However, no significant difference was observed in TPAC between the two groups studied. The results of this preliminary study suggest that from the five activation receptors and co-receptors of NK cells evaluated (NKp30, NKp46, NKp80, NKG2D and 2B4), only NKp30 and NKG2D receptor expression was diminished in women exposed to toxics when compared with data from unexposed women. These results suggest that the occupational exposure to mixture of toxics is one of the important factors in the diminution of the NK cell receptor expression.
IL-21 augments natural killer effector functions in chronically HIV infected indi-viduals doi:10.1097/QAD.0b013e3283089367 stimulation of nk cells for cancer patients Copyright
  • Armas N L Strbo
  • H Liu
  • Ma Kolber
  • M Lichtenheld
  • Pahwa
Strbo N, Armas L, Liu H, Kolber MA, Lichtenheld M, Pahwa S. IL-21 augments natural killer effector functions in chronically HIV infected indi-viduals. AIDS 2008;22:1551–1560. doi:10.1097/QAD.0b013e3283089367 stimulation of nk cells for cancer patients Copyright © 2013 John Wiley & Sons, Ltd. Cell Biochem Funct (2013)
Strategies of NK cell recognition and their roles in tumor immunosurveillance. In How the Immune System Recognizes Self and Nonself: Immunoreceptors and Their Signaling
  • Ca Stewart
  • Vivier
Stewart CA, Vivier E. Strategies of NK cell recognition and their roles in tumor immunosurveillance. In How the Immune System Recognizes Self and Nonself: Immunoreceptors and Their Signaling. Kitamura D (ed.). Springer: Tokyo, Japan, 2007; 37–81.
Intraepithelial CD8+ tumor-infiltrating lymphocytes and a high CD8/regulatory T cell ratio are associated with favorable prognosis in ovarian cancer
  • Sato
Sato E, Olson SH, Ahn J, et al. Intraepithelial CD8+ tumor-infiltrating lymphocytes and a high CD8/regulatory T cell ratio are associated with favorable prognosis in ovarian cancer. Proc Natl Acad Sci U S A. 2005;102(51):18538-18543. doi:10.1073/pnas.0509182102