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Modulation of Inflammatory Effects of Cytokines by the Neuropeptide α-MSH
Published in Sami I. Said, Proinflammatory and Antiinflammatory Peptides, 2020
J. M. Lipton, Nilum Rajora, R. A. Star, S. Taherzadeh, Anna Catania, Giuliana Ceriani, G. Boccoli
Although cultured RAW cells share characteristics of normal human macrophages, there is controversy concerning human monocyte/macrophage production of nitric oxide. The THP-1 cell line, a human monocytic leukemia cell line, can be induced to differentiate into macrophages by the phorbol ester PM A. This cell line resembles normal human monocytes with regard to morphology, secretory products, oncogene expression, expression of membrane antigens, and expression of genes involved in lipid metabolism. To determine if the antiinflammatory effects of α-MSH obtained with murine macrophages also occur in cells of human monocyte/macrophage characteristics, a series of in-vitro tests were performed using THP-1 cells (34). These cells produce neopterin, and α-MSH inhibited production of this primate homolog of cytotoxic nitric oxide. It thus appears that α-MSH inhibits inflammatory products in human macrophages much as it does in murine macrophages. Modulation by α-MSH of specific inflammatory mediators/cytotoxic agents appears to differ depending on the importance of the mediators in myelomonocytic cells of different species.
Immediate Cytokine Responses to Endotoxin: Tumor Necrosis Factor-α and the lnterleukin-1 Family
Published in Helmut Brade, Steven M. Opal, Stefanie N. Vogel, David C. Morrison, Endotoxin in Health and Disease, 2020
As shown in Figure 1, there is an ordered stimulation of TNF and IL-1 gene expression. TNF is clearly a very early gene in monocytes after exposure to LPS. In most studies, human peripheral blood mononuclear cells (PBMC) are incubated with low concentrations of LPS (1–10 ng/ml) and within 10–15 minutes there is an increase in the steady-state levels of TNF-α mRNA. After 3–4 hours these are dramatically reduced. IL-1β is slower in onset compared to TNF-α. One usually does not observe a significant increase in IL-1β steady-state mRNA levels until 3 minutes, but the expression is longer and more sustained than that for TNF-α. In the THP-1 cell line, the expression of IL-1β has been studied in detail (10). Both TNF-α and IL-1β have a 3’ untranslated sequence that effects mRNA stability (11). This region is thought to be responsive to LPS effects.
Enzyme-Releasing Peptide
Published in Jason Kelley, Cytokines of the Lung, 2022
Allen B. Cohen, Edmund J. Miller, Cassandra MacArthur
Recent results have shown that ERP is likely to be synthesized by the macrophage precursor cell lines, THP-1, HL-60, and U937, but not by human alveolar macrophages (MacArthur et al., 1991). All three of these cell lines are derived from cell lines that can be considered to be macrophage precursors. U937 and HL-60 are derived from promyelocytic leukemic cell lines (Gallagher et al., 1979; Collins, 1987; Harris and Ralph, 1985), and the THP-1 cell line was developed from a child with monocytic leukemia (Tsuchiya 1980, 1982). The THP-1 cells secreted more ERP than human alveolar macrophages. The THP-1 cells secreted 12.3 nM/18 h per 5 × 106 cells, and macrophages secreted 3.6 nM/18h per 5 × 106 cells. The ERP secreted by the THP-1 cells was an active neutrophil secretagogue at concentrations of less than 1 µM. A western blot analysis of the THP-1–conditioned medium indicated that a high and a low Mr form of ERP existed (Fig. 5). Only the low Mr form of ERP released enzymes from neutrophils. Incubation of human alveolar macrophages with macrophage stimulants or with inhibitors of protein synthesis did not modify the secretion of ERP into the medium (Fig. 6). However, incubation of the THP-1 cells with the same reagents resulted in significant inhibition in the secretion of ERP by dactinomycin (actinomycin D), two concentrations of cycloheximide, and phorbol esters (Fig. 7). Fluorescence microscopy of the THP-1 cells showed peripheral fluorescence in unstimulated cells.
Cytotoxic effects of extracts obtained from plants of the Oleaceae family: bio-guided isolation and molecular docking of new secoiridoids from Jasminum humile
Published in Pharmaceutical Biology, 2022
Khaled Ahmed Mansour, Ahmed Elbermawi, Ahmed A. Al-Karmalawy, Mohamed-Farid Lahloub, Mona El-Neketi
The activity-guided chromatographic isolation led to identifying five compounds (1–4 and 6) from the ethyl acetate fraction and two compounds from the n-butanol fraction (5 and 7). All isolated compounds were investigated for their cytotoxic activity (Table 2). Different concentrations were used and statistical significance was determined compared to the reference drug doxorubicin. Compound (5) showed the highest effect against THP-1 cell line. It is evident that it is very promising for continued research as an anticancer agent due to its high potency (66.47 μg/mL against HepG-2, IC50 of 41.32 μg/mL against MCF-7, and 27.59 μg/mL against THP-1) as well as high selectivity (SI = 3.16 for HepG-2, 5.09 for MCF-7, and 7.62 for THP-1) on the tested cell lines (Table 2).
In vitro OP9-DL1 co-culture and subsequent maturation in the presence of IL-21 generates tumor antigen-specific T cells with a favorable less-differentiated phenotype and enhanced functionality
Published in OncoImmunology, 2021
Sarah Bonte, Stijn de Munter, Lore Billiet, Glenn Goetgeluk, Joline Ingels, Hanne Jansen, Melissa Pille, Laurenz de Cock, Karin Weening, Tom Taghon, Georges Leclercq, Bart Vandekerckhove, Tessa Kerre
The T2 cell line, a human HLA-A2+ cell line deficient for transporter associated with antigen processing (TAP), and JY cell line, a human HLA-A2+ EBV-immortalized B-cell line, were obtained from the American Type Culture Collection (ATCC) and cultured in Iscove’s Modified Dulbecco’s Medium (IMDM; Thermo Fisher Scientific, 12440053) supplemented with 10% fetal calf serum (FCS), 2 mM L-glutamine (Thermo Fisher Scientific, 25030–081), 100 IU/ml penicillin and 100 IU/ml streptomycin (Thermo Fisher Scientific, 15140–122) (complete IMDM, cIMDM). The HL-60-A2 cell line, a kind gift from Dr B. Depreter (University Hospital Brussels, Belgium;19), was also cultured in cIMDM. The THP-1 cell line, a human monocytic cell line derived from a patient with acute monocytic leukemia, a kind gift from Dr N. Lambrechts (VITO, Mol, Belgium), was cultured in Roswell Park Memorial Institute medium (RPMI; Thermo Fisher Scientific, 52400–025) supplemented with 10% FCS, 2 mM L-glutamine, 100 IU/ml penicillin, 100 IU/ml streptomycin and 0.05 mM 2-mercaptoethanol (Sigma-Aldrich, M6250). The OP9-DL1 cell line, a mouse stromal cell line genetically modified to express the human Notch ligand Delta-like 1 (DL1), was a kind gift from Dr J. C. Zúñiga-Pflücker (University of Toronto, Canada). This cell line was cultured in Minimum Essential Medium alpha (MEMα; Thermo Fisher Scientific, 12561–056) supplemented with 20% FCS, 2 mM L-glutamine, 100 IU/ml penicillin and 100 IU/ml streptomycin. OP9-DL1 cells were grown near confluence, harvested and split every 2–3 days.
Mannosylated solid lipid nanoparticles for the selective delivery of rifampicin to macrophages
Published in Artificial Cells, Nanomedicine, and Biotechnology, 2018
Alexandre C. C. Vieira, Luíse L. Chaves, Marina Pinheiro, Sofia A. Costa Lima, Domingos Ferreira, Bruno Sarmento, Salette Reis
RIF (more than 98% pure), stearylamine (97% pure) and D-(+)-Mannose (more than 99% pure) were purchased from Sigma-Aldrich (St Louis, MO, USA). Aerosil® 200 was supplied by Acofarma® (Terrassa, Espanha, Spain.) The SLNs were prepared with glycerol tripalmitate provided by Alfa Aesar (Karlsruhe, Germany), Tween® 80 purchased from Sigma-Aldrich Co. (St Louis, MO, USA). Phorbol 12-myristate 13-acetate (PMA), trypan blue, thiazolyl blue tetrazolium bromide (MTT), phosphate buffered saline pH 7.4 (PBS), sucrose, chlorpromazine hydrochloride, filipin, cytochalasin D and coumarin 6 were obtained from Sigma–Aldrich (St. Louis, MO, USA). All other chemicals used in the study were of analytical grade. Dulbecco’s Modified Eagle’s Medium (DMEM), Fetal Bovine Serum (FBS) and penicillin-streptomycin antibiotics mixture were purchased from Gibco® (Invitrogen Corporation, London, UK). Human leukemia monocyte THP1 cell line from European Culture Collections (Salisbury, UK).