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Frontiers in Immunology 2022Cell-based functional immune-assays may allow for risk stratification of patients with complex, heterogeneous immune disorders such as sepsis. Given the heterogeneity of... (Observational Study)
Observational Study
BACKGROUND
Cell-based functional immune-assays may allow for risk stratification of patients with complex, heterogeneous immune disorders such as sepsis. Given the heterogeneity of patient responses and the uncertain immune pathogenesis of sepsis, these assays must first be defined and calibrated in the healthy population.
OBJECTIVE
Our objective was to compare the internal consistency and practicality of two immune assays that may provide data on surrogate markers of the innate and adaptive immune response. We hypothesized that a rapid turnaround, microfluidic-based immune assay (ELLA) would be comparable to a dual-color, enzyme-linked immunospot (ELISpot) assay in identifying tumor necrosis factor (TNF) and interferon (IFN)γ production following whole blood stimulation.
DESIGN
This was a prospective, observational cohort analysis. Whole blood samples from ten healthy, immune-competent volunteers were stimulated for either 4 hours or 18 hours with lipopolysaccharide, anti-CD3/anti-CD28 antibodies, or phorbol 12-myristate 13-acetate with ionomycin to interrogate innate and adaptive immune responses, respectively.
MEASUREMENTS AND MAIN RESULTS
ELLA analysis produced more precise measurement of TNF and IFNγ concentrations as compared with ELISpot, as well as a four- to five-log dynamic range for TNF and IFNγ concentrations, as compared with a two-log dynamic range with ELISpot. Unsupervised clustering accurately predicted the immune stimulant used for 90% of samples analyzed ELLA, as compared with 72% of samples analyzed ELISpot.
CONCLUSIONS
We describe, for the first time, a rapid and precise assay for functional interrogation of the innate and adaptive arms of the immune system in healthy volunteers. The advantages of the ELLA microfluidic platform may represent a step forward in generating a point-of-care test with clinical utility, for identifying deranged immune phenotypes in septic patients.
Topics: Cytokines; Enzyme-Linked Immunospot Assay; Humans; Interferon-gamma; Prospective Studies; Sepsis; Tetradecanoylphorbol Acetate; Tumor Necrosis Factor-alpha
PubMed: 35860253
DOI: 10.3389/fimmu.2022.940030 -
Journal of Natural Products May 2023Three new phenanthrene derivatives (, , ), one new fluorenone (), and four known compounds (-) were isolated from the ethyl acetate extract of Sw. stems using column...
Three new phenanthrene derivatives (, , ), one new fluorenone (), and four known compounds (-) were isolated from the ethyl acetate extract of Sw. stems using column chromatography. The chemical structures were elucidated by analysis of spectroscopic data. The absolute configuration of was determined by electronic circular dichroism calculation. We also evaluated the immunomodulatory effects of compounds isolated from in human peripheral blood mononuclear cells from healthy individuals and those from patients with multiple sclerosis in vitro. Dendrocrumenol B () and dendrocrumenol D () showed strong immunomodulatory effects on both CD3 T cells and CD14 monocytes. Compounds and could reduce IL-2 and TNF production in T cells and monocytes that were treated with phorbol-12-myristate-13-acetate and ionomycin (PMA/Iono). Deep immune profiling using high-dimensional single-cell mass cytometry could confirm immunomodulatory effects of , quantified by the reduction of activated T cell population under PMA/Iono stimulation, in comparison to the stimulated T cells without treatment.
Topics: Humans; Dendrobium; Leukocytes, Mononuclear; Monocytes; Phenanthrenes; T-Lymphocytes; Tetradecanoylphorbol Acetate; Fluorenes
PubMed: 37140218
DOI: 10.1021/acs.jnatprod.3c00107 -
Natural Product Research Oct 2022Five tigliane (-) and one ingenane () diterpenoids were isolated from the ethanol extract of . The structures of these compounds were identified based on analysis of...
Five tigliane (-) and one ingenane () diterpenoids were isolated from the ethanol extract of . The structures of these compounds were identified based on analysis of their spectroscopic data. Among them, compound 12--(2,4,6,8-tetradecatetraenoyl)-13--isobutyroyl-4-deoxyphorbol () was a new tigliane. The Rho123 effluxion assay showed that tiglianes with a -fused 5/7 ring system such as compounds , , and had potent inhibitory activity against P-glycoprotein in HepG2/ADR cells.
Topics: ATP Binding Cassette Transporter, Subfamily B; Diterpenes; Ethanol; Euphorbia; Molecular Structure; Phorbols; Plant Extracts
PubMed: 34142618
DOI: 10.1080/14786419.2021.1938039 -
Cancer Prevention Research... Jan 2015Consuming plants for their presumed health benefits has occurred since early civilizations. Phytochemicals are found in various plants that are frequently included in... (Review)
Review
Consuming plants for their presumed health benefits has occurred since early civilizations. Phytochemicals are found in various plants that are frequently included in the human diet and are generally thought to be safe for consumption because they are produced naturally. However, this is not always the case and in fact many natural compounds found in several commonly consumed plants are potential carcinogens or tumor promoters and should be avoided.
Topics: Amygdalin; Aristolochic Acids; Capsaicin; Cell Line, Tumor; Cycasin; Diet; Drug Screening Assays, Antitumor; Humans; Indans; Neoplasms; Phorbol Esters; Phytochemicals; Phytoestrogens; Phytotherapy; Plant Extracts; Plants; Pyrrolizidine Alkaloids; Safrole; Sesquiterpenes
PubMed: 25348854
DOI: 10.1158/1940-6207.CAPR-14-0160 -
Journal of Natural Products Jun 2019Chemical investigation of an EtOH extract of the twigs and leaves of Croton damayeshu afforded 10 new tigliane diterpenoids, crodamoids A-J (1-10), along with five known...
Chemical investigation of an EtOH extract of the twigs and leaves of Croton damayeshu afforded 10 new tigliane diterpenoids, crodamoids A-J (1-10), along with five known compounds. Their structures were elucidated by physical data analysis. Compounds 8, 9, and 15 displayed cytotoxic effects against two human tumor cell lines, A549 and HL-60 (IC: 0.9-2.4 μM).
Topics: Antineoplastic Agents, Phytogenic; Cell Line, Tumor; Croton; Diterpenes; HL-60 Cells; Humans; Magnetic Resonance Spectroscopy; Molecular Structure; Phorbols; Plant Leaves
PubMed: 31117522
DOI: 10.1021/acs.jnatprod.9b00042 -
Journal of Pharmacological Sciences Oct 2022Non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases, and there are no effective drugs available so far. Lactucin and Lactucopicrin...
Non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases, and there are no effective drugs available so far. Lactucin and Lactucopicrin belong to sesquiterpene lactones and are extracted from Cichorium glandulosum Boiss. et Huet (CG) possesses multiple biopharmacological activities. However, the therapeutic effects of both Lactucin and Lactucopicrin on many diseases and their underlying mechanisms remain largely unknown. Here, we analyzed the both natural compounds hypolipidemic effects on FFA-induced HepG2 cells and their potential mechanisms based on transcriptomics and experimental tests. Our results indicated that Lactucin (10 μM) and Lactucopicrin (20 μM) remarkably reduced TG accumulation. Transcriptomics analysis identified 1960, 1645, and 1791 differentially expressed genes (DEGs) and obtained 611 and 635 specific genes in different comparisons, respectively. The enrichment analysis and experimental validations (RT-qPCR and Western Blot) showed that their hypolipidemic activities were most probably exerted via regulating numerous key DEGs involved in lipid metabolism. Taken together, both Lactucin and Lactucopicrin may represent potent hepatoprotective agents. Both of them exhibited therapeutic effects against liver diseases such as NAFLD by regulating multi-gene and proteins like HADHA, ADAM17, SQSTM1, and GBA and modulating multi-pathways like fatty acid oxidation metabolic signaling.
Topics: Hep G2 Cells; Humans; Lactones; Lipid Metabolism; Non-alcoholic Fatty Liver Disease; Phorbols; Sesquiterpenes
PubMed: 36055749
DOI: 10.1016/j.jphs.2022.07.007 -
Journal of Natural Products Feb 2022Thirty-five tigliane diterpenoids and two -kaurane diterpenoids were isolated from the leaves of , and, among them, compounds - were characterized as new tigliane...
Thirty-five tigliane diterpenoids and two -kaurane diterpenoids were isolated from the leaves of , and, among them, compounds - were characterized as new tigliane diterpenoids. The structures of compounds - were determined by analysis of their HRESIMS, NMR, and ECD data and by chemical methods. The isolates were assayed for their larvicidal, antifungal, and α-glucosidase inhibitory activities, and compounds - were found to possess larvicidal activities against with LC values of 0.19, 0.16, and 0.26 μM, respectively, comparable to the LC of 0.14 μM for the positive control, flubendiamide.
Topics: Antifungal Agents; Croton; Diterpenes; Diterpenes, Kaurane; Molecular Structure; Phorbols; alpha-Glucosidases
PubMed: 35080403
DOI: 10.1021/acs.jnatprod.1c00977 -
Bioorganic & Medicinal Chemistry Letters Oct 2021Tigliane esters show many biological activities, including anti-HIV-1 activity. Our aim in this study was to establish structure-anti-HIV activity relationships for four...
Tigliane esters show many biological activities, including anti-HIV-1 activity. Our aim in this study was to establish structure-anti-HIV activity relationships for four series of tigliane-type diterpenoids. We synthesized and evaluated 29 new phorbol ester derivatives for anti-HIV activity and for cytotoxicity against human tumor cell lines. Among them, three derivatives, two phorbol-13-monoesters (5d and 5e) and a phorbol-12,13-diester (6a), showed significant anti-HIV activity. We found that better anti-HIV activity was often associated with a shorter acyl ester at C-13. Particularly, compounds with a phenyl ring in the ester side chain exhibited excellent anti-HIV activity and had good safety indexes. Due to its significant anti-HIV potency with a high selectivity index, phorbol-12,13-dicinnamoate (6a) was chosen as the potential candidate for further preclinical trials.
Topics: Anti-HIV Agents; Antineoplastic Agents; Cell Line, Tumor; Cell Survival; HIV-1; Humans; Molecular Structure; Phorbol Esters; Structure-Activity Relationship; Virus Replication
PubMed: 34403728
DOI: 10.1016/j.bmcl.2021.128319 -
Bulletin of Experimental Biology and... Apr 2021We analyzed functional status of blood leukocytes in diabetes mellitus and after addition of glucose in vitro. To this end, generation of ROS and reactive halogen...
We analyzed functional status of blood leukocytes in diabetes mellitus and after addition of glucose in vitro. To this end, generation of ROS and reactive halogen species by monocytes and neutrophils from patients with diabetes mellitus and healthy donors was assayed using lucigenin- and luminol-dependent chemiluminescence after stimulation with phorbol 12-myristate 13-acetate or opsonized zymosan in vitro. Formation of neutrophil extracellular traps was evaluated in the blood after addition of glucose. In comparison with donors, leukocytes from patients with diabetes mellitus were primed and this effect can be modeled by addition of glucose to the blood in vitro. Addition of glucose to donor blood also triggered the formation of neutrophil extracellular traps.
Topics: Diabetes Mellitus; Extracellular Traps; Humans; Hyperglycemia; Leukocytes; Luminescent Measurements; Neutrophils; Reactive Oxygen Species; Tetradecanoylphorbol Acetate; Zymosan
PubMed: 33893954
DOI: 10.1007/s10517-021-05147-x -
IUBMB Life Jun 2019Protein kinase C (PKC) is activated by 1,2-diacylglycerol as a second messenger in the signaling mechanism coupled with the hydrolysis of membrane inositol... (Review)
Review
Protein kinase C (PKC) is activated by 1,2-diacylglycerol as a second messenger in the signaling mechanism coupled with the hydrolysis of membrane inositol phospholipids, although it was not found by screening for a 1,2-diacylglycerol-dependent enzyme. PKC is also a receptor for the tumor-promoting phorbol esters, but it was not identified by its property of binding phorbol esters, either. Instead, the discovery and characterization of PKC, now known to comprise a family with multiple isoforms, was through a circuitous voyage filled with unexpected twists and turns. This review summarizes the discovery and the initial experiments of PKC as a historical perspective of the enzyme family in the context of the progress in the studies on protein phosphorylation. © 2018 IUBMB Life, 71(6):697-705, 2019.
Topics: Diglycerides; Humans; Hydrolysis; Phorbol Esters; Phosphatidylinositols; Phosphorylation; Protein Binding; Protein Kinase C; Proteins
PubMed: 30393952
DOI: 10.1002/iub.1963