There are substances that kill cancer cells, but induce T cell proliferation, like thalidomide. To find more of these, a new anticancer drug screening strategy is vital. In this study we report the development of a differential cytotoxicity screening or evaluation platform using the CellTiter-Glo (Promega, Annandale, NSW, Australia) luminescent cell viability assay (ATP assay) and also the CellTiter 96 AQueous (Promega) one solution cell proliferation assay [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay]. The results suggested the platform consisting of the combination of the ATP assay be used for quantifying peripheral blood mononuclear cells, while the more economic MTS colorimetric assay is well suited to be used detecting cell viability of cancer cells. In addition, we found paclitaxel (Taxol, MP Biomedicals Australasia Pty Ltd., Seven Hills, NSW, Australia) to be a useful control for this routine screening methodology. Taxol exhibits the desirable in vitro feature of differential cytotoxicity that spares the immunological cells, when used at a concentration that will kill the majority of the cancer cell population.
Polysaccharopeptide (PSP), from Coriolus versicolor, has been used widely as an adjuvant to chemotherapy with demonstrated anti-tumor and broad immunomodulating effects. While PSP’s mechanism of action still remains unknown, its enhanced immunomodulatory potential with acacia gum is of great interest. Acacia gum, which also contains polysaccharides and glycoproteins, has been demonstrated to be immunopotentiating. To elucidate whether PSP directly activates T-cell-dependent B-cell responses in vivo, we used a well-established hapten carrier system (Nitrophenyl-chicken gamma globulin (NP-CGG)). 6-week C57BL/6 male mice were immunised with 50 μg of NP25-CGG alum precipitate intraperitoneally. Mice were gavaged daily with 50mg/kg PSP in a vehicle containing acacia gum and sacrificed at days 0, 4, 7, 10, 14 and 21. ELISA was used to measure the total and relative hapten-specific anti-NP IgA, IgM and IgG titre levels compared to the controls. It was found that PSP, combined with acacia gum, significantly increased total IgG titre levels at day 4 (P< 0.05), decreased IgM titre levels at days 4 and 21 (P< 0.05) with no alterations observed in the IgA or IgE titre levels at any of the time points measured. Our results suggest that while PSP combined with acacia gum appears to exert weak immunological effects through specific T-cell dependent B-cell responses, they are likely to be broad and non-specific which supports the current literature on PSP. We report for the first time the application of a well-established hapten-carrier system that can be used to characterise and delineate specific T-cell dependent B-cell responses of potential immunomodulatory glycoprotein-based herbal medicines combinations in vivo.
Background: AL is a traditional Chinese medicine (TCM) combination commonly used as an adaptogen to improve energy levels, immunity and quality of life in those suffering from chronic diseases such as cancer. While the combination Astragalus membranaceus and Ligustrum lucidum (AL) has been investigated in China as an adjunct to standard anticancer therapies in numerous clinical studies of variable quality reported in the Chinese literature, independent assessment of its effects on safety, tolerability and efficacy are lacking. Our objectives are to determine the safety and tolerability of AL and investigate its effects on inflammation, quality of life and immunity in people with advanced malignancy in the Australian healthcare setting. Methods: The AL study is a prospectively registered, open labelled pilot multi-centre study investigating AL in people with advanced malignancy. Inclusion criteria include participants with recurrent or metastatic cancer who are not undergoing chemotherapy or palliative chemotherapy. All participants (n=25) will receive 6 capsules of AL twice daily (equivalent to 25g raw herb) for 12 weeks. Follow up consultations will monitor safety, tolerability, quality of life, immune function and adverse events. Participants will be assessed at baseline and at weeks 3, 6, 9 and 12. The primary outcome will determine the effect of AL on safety and tolerability. Secondary outcomes will include inflammation, quality of life, immune function, disease status and survival. Appropriate statistical analysis will be conducted on the pilot study data. Potential associations will be investigated where relevant. Conclusions: This study will firstly establish the safety and tolerability of this TCM combination "AL" in people with advanced malignancy in the Australian healthcare system and provide important information regarding its effect on markers that may affect survival as well as explore changes in quality of life and immune function. The impact of this research may allow the design of future studies integrating AL with standard therapy for people with advanced malignancy.
One fraction may potentially contain valuable compounds which may be useful for further investigation. This may focus on the identification of the bioavailable purified compounds present within these fractions or by detailed delineation of the related mechanisms of action.
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