scholarly journals Differential Cytokine Response in Host Defence Mechanisms Triggered by Gram-Negative and Gram-Positive Bacteria, and the Roles of Gabexate Mesilate, a Synthetic Protease Inhibitor

2002 ◽  
Vol 30 (2) ◽  
pp. 99-108 ◽  
Author(s):  
H Iwadou ◽  
Y Morimoto ◽  
H Iwagaki ◽  
S Sinoura ◽  
Y Chouda ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Bacterial Infections ◽  
Mononuclear Cells ◽  
Cytokine Production ◽  
Gabexate Mesilate ◽  
Gram Positive ◽  
Peripheral Blood Mononuclear ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Tnf Α

Bacterial infection results in the production of inflammatory mediators and may be involved in the pathogenesis of sepsis and/or systemic inflammatory response syndrome. The effect of lipopolysaccharide (LPS), a major component of the outer surface of Gram-negative bacteria, and Staphylococcal enterotoxin B (SEB), a superantigen of Gram-positive bacteria, on cytokine production in peripheral blood mononuclear cells (PBMCs) was examined. LPS significantly increased the production of proinflammatory and anti-inflammatory cytokines, and SEB enhanced the production of helper T lymphocyte type cytokines. These results illustrated the different responses to Gram-negative and Gram-positive bacterial infections. The effect of gabexate mesilate, a synthetic protease inhibitor, on cytokine production and expression of the toll-like receptor (TLR) was also examined. The results suggest that gabexate mesilate-induced inhibition of tumour necrosis factor-α (TNF-α) and interleukin-18 (IL-18) production in LPS-stimulated PBMCs is due to the inhibition of the nuclear factor-κB activation pathway and/or inhibition of the processing pathway of pro-TNF-α and pro-IL-18, not to down-regulation of TLR-2 or TLR-4.

scholarly journals Innate Immune Responses of Human Neonatal Cells to Bacteria from the Normal Gastrointestinal Flora

2002 ◽  
Vol 70 (12) ◽  
pp. 6688-6696 ◽  
Author(s):  
Helen Karlsson ◽  
Christina Hessle ◽  
Anna Rudin
Keyword(s):  
Immune System ◽  
Mononuclear Cells ◽  
Innate Immune ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bacterial Strains ◽  
Gram Positive ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Tnf Α ◽  
Adult Cells

ABSTRACT The hygiene hypothesis postulates that the prevalence of allergy has increased due to decreased microbial stimulation early in life, leading to delayed maturation of the immune system. The aim of this study was to examine the cytokine pattern produced from cord blood mononuclear cells relative to adult cells after stimulation with bacterial strains from the normal flora. Mononuclear cells from cord and adult blood samples were stimulated with the following bacteria: Bifidobacterium adolescentis, Enterococcus faecalis, Lactobacillus plantarum, Streptococcus mitis, Corynebacterium minutissimum, Clostridium perfringens, Bacteroides vulgatus, Escherichia coli, Pseudomonas aeruginosa, Veillonella parvula, and Neisseria sicca. The levels of interleukin 12 (IL-12), tumor necrosis factor alpha (TNF-α), IL-10, and IL-6 were measured by enzyme-linked immunosorbent assay. The TNF-α production was also analyzed after blocking CD14, Toll-like receptor 2 (TLR-2), and TLR-4 prior to stimulation with bacteria. The levels of IL-12 and TNF-α were similar in cord and adult cells. Gram-positive bacteria induced considerably higher levels of IL-12 and TNF-α than gram-negative bacteria in both cord and adult cells. The levels of IL-6 were significantly higher in newborns than in adults, whereas the levels of IL-10 were similar in newborns and adults. Gram-negative and gram-positive bacteria induced similar levels of IL-6 and IL-10 in cord cells. L. plantarum bound or signaled through CD14, TLR-2, and TLR-4, whereas E. coli acted mainly through CD14 and TLR-4. These results indicate that the innate immune response in newborns to commensal bacteria is strong and also suggest that different bacterial strains may have differential effects on the maturation of the immune system of infants.

Dysfunction of Toll-Like Receptor-2, −4, and −6: Is There a Role in the Pathogenesis of Immunodeficiency in Multiple Myeloma?.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3430-3430
Author(s):  
Evangelos Terpos ◽  
Meletios A. Dimopoulos ◽  
Charoula Xirakia ◽  
Dimitrios Christoulas ◽  
Athanasios Anagnostopoulos ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bacterial Infections ◽  
Mononuclear Cells ◽  
Healthy Controls ◽  
Limited Information ◽  
Becton Dickinson ◽  
Gram Positive ◽  
Peripheral Blood Mononuclear ◽  
Tnf Α ◽  
And Gender

Abstract Infections are a major cause of death in multiple myeloma (MM). There is limited information for the possible role of innate immunity in the pathogenesis of immunodeficiency in MM. Innate immune detection of pathogens relies on specific classes of microbial sensors, such as Toll-like receptors (TLRs) that detect structural patterns, which do not exist in the host. The aim of this study was to evaluate the expression and function of TLRs in newly diagnosed patients with MM. According to our knowledge, such information is not available in the literature. Thirty-two MM patients at diagnosis (18M/14F; median age 69 years), 6 MGUS patients and 14 healthy, age- and gender-matched controls were studied. Seven patients had stage 1, 14 stage 2 and 11 stage 3 myeloma, according to ISS. After the collection of peripheral blood, mononuclear cells (PBMCs) were isolated by Ficoll centrifugation (Histopaque-1077; Sigma-Aldrich). These cells were measured for the expression of TLRs (antibodies from eBioscience) using fluorescence activated flow cytometry (FC 500, Beckman Coulter). In addition, 1x106 cells/ml were cultured in 5% FCS 1% pen/strep RPMI in the presence or absence of various TLR ligands and supernatants collected after 20h. These were examined for the presence of inflammatory cytokines (tumor necrosis-alpha, TNF-α; and interleukin-6, IL-6) by ELISA (Becton Dickinson). We found that although patients with MM express TLRs in PBMCs, their response to certain TLR ligands is defective when compared to healthy controls. TLR2, TLR4 and TLR6 of PBMCs of healthy controls reacted to the presence of their respective ligands PAM3CYS (gram positive and negative bacterial), LPS (gram negative) and FLT-1 (gram positive) producing TNF-α at a median value of 2.4 ng/mL (range: 1–3 ng/ml), while their action in patients with MM was significantly reduced (median value of TNF-α: 190 pg/ml; range 100–500 pg/ml; p<0.001). The reduced response of TLR4 in MM patients was independent of the LPS concentration used. On the contrary, TLR7 and TLR8 from MM reacted normally to their ligand R-848 (Imiquimob) to secrete high levels of TNF-α (median value for patients and controls: 4.5 and 4.2 ng/ml, respectively; p=NS). NOD1, another pattern recognition receptor that recognizes bacterial peptidoglycans also reacted normally in MM. Similar observations have been made for IL-6 expression. There was no difference in terms of TLRs function between MGUS patients and controls and between myeloma patients of different disease stages. In seven MM patients, cells were pre-treated with bortezomib for 30 min before addition of the TLR ligand. Bortezomib administration abrogated TLR 7/8 response in PBMCs even at very low concentrations (1nM). This study suggests that there is a significant defect in TLR function in MM, especially of these involved in immunity against bacterial infections. Thus the immune system fails to receive early priming signal, which may contribute to the increased rate of infections observed in MM. The restoration of function of TLRs to their normal levels has the potential to improve bacterial immunity in MM.

scholarly journals In Vitro Antitumor Activity of Endophytic and Rhizosphere Gram-Positive Bacteria from Ibervillea sonorae (S. Watson) Greene against L5178Y-R Lymphoma Cells

2022 ◽  
Vol 19 (2) ◽  
pp. 894
Author(s):  
Ricardo Romero-Arguelles ◽  
César Iván Romo-Sáenz ◽  
Karla Morán-Santibáñez ◽  
Patricia Tamez-Guerra ◽  
Ramiro Quintanilla-Licea ◽  
...  
Keyword(s):  
Antitumor Activity ◽  
Growth Inhibition ◽  
Mononuclear Cells ◽  
Colorimetric Assay ◽  
Human Peripheral Blood ◽  
Antitumor Effects ◽  
Lymphoma Cells ◽  
Gram Positive ◽  
Peripheral Blood Mononuclear ◽  
Gram Positive Bacteria

Plant-associated microorganisms represent a potential source of new antitumor compounds. The aim of the present study was to isolate endophytic and rhizosphere Gram-positive bacteria from Ibervillea sonorae and produce extracts with antitumor activity. Methanol and ethyl acetate extracts were obtained from 28 d bacterial fermentation, after which murine L5178Y-R lymphoma cells growth inhibition was evaluated at concentrations ranging from 15.62 µg/mL to 500 µg/mL by the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide reduction colorimetric assay. IC50 and the selectivity index (SI) were calculated and compared with healthy control human peripheral blood mononuclear cells (PBMC). Identification of the isolated strains was performed using the 16S ribosomal gene and by MALDI-TOF MS mass spectrometry. The endophytic and rhizosphere bacterial extracts from strains ISE-B22, ISE-B26, ISE-B27, ISS-A01, ISS-A06, and ISS-A16 showed significant (p < 0.05) L5178Y-R cell growth inhibition, compared with an untreated control. The rhizosphere Micromonospora echinospora isolate ISS-A16 showed the highest (90.48%) percentage of lymphoma cells growth inhibition and SI (19.1) for PBMC, whereas the Bacillus subtilis ISE-B26 isolate caused significant (p < 0.01) growth inhibition (84.32%) and a SI of 5.2. Taken together, results of the present study evidenced antitumor effects by I. sonorae endophytic and rhizosphere bacteria culture extracts. Further research will involve the elucidation of the compounds that exert the antitumor activity and their evaluation in pre-clinical studies.

Polymyxin-B Stimulates Tumor Necrosis Factor-α Production by Human Peripheral Blood Mononuclear Cells

1998 ◽  
Vol 21 (5) ◽  
pp. 269-273 ◽  
Author(s):  
B.L. Jaber ◽  
S. Sundaram ◽  
M. Cendoroglo Neto ◽  
A.J. King ◽  
B.J.G. Pereira
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Tumor Necrosis ◽  
Mononuclear Cells ◽  
Cytokine Production ◽  
Polymyxin B ◽  
Peripheral Blood Mononuclear ◽  
Gram Negative ◽  
Blood Mononuclear Cells

Gram-negative bacterial lipopolysaccharide (LPS) is a well known stimulus for cytokine production, particularly interleukin-1 (IL-1) and tumor necrosis tactor alpha (TNFα). Polymyxin B (PMX-B) is a cationic polypeptide that binds to LPS, neutralizing its biological effects. PMX-B also disrupts gram-negative bacterial cell membrane phospholipids but is highly toxic to mammalian cells, therefore is of limited use. PMX-B is used as additive to media, as a way to handle LPS contamination. To derive benefit from the ability of PMX-B to neutralize lipid A in vivo while avoiding its systemic toxicity, PMX-B was covalently bound to polystyrene-derivative fibers, creating a hemoperfusion column (PMX-F) for the selective removal of circulating ET In vitro PMX-F hemoperfusion studies have demonstrated effective ET removal, using either the Limulus amebocyte lysate assay or TNFα production by peripheral blood mononuclear cells (PBMC) as an index of ET removal. However, the question whether PMX-B itself could stimulate human PBMC to produce cytokines has not been adequately addressed. We examined the effect of increasing concentrations of PMX-B on cytokine production by PBMC in vitro. PBMC harvested from healthy volunteers were incubated for 24 hours at 37°C with control (tissue culture media RPMI), or 5 µg/ml, 10 µg/ml, 20 µg/ml or 100 µg/ml PMX-B. At the end of 24 hours, PBMC were subjected to three freeze-thaw cycles, and total TNFα production (pg/2.5x106 PBMC) was measured by radioimmunoassay. Total TNFα production by PBMC was 163 ± 3 pg, 171 ± 9 pg, 164 ± 4 pg, 323 ± 63 pg and 331 ± 58 pg, in the control, PMX-B 5 µg/ml, 10 µg/ml, 20 µg/ml and 100 µg/ml conditions, respectively. Compared to controls (RPMI), the percentage increase in TNFα production by PBMC was 5 ± 6% (P=0.23), 1 ± 3% (P=0.45), 99 ± 40% (P=0.03) and 103 ± 36% (P=0.02) in the presence of 5 µg/ml, 10 µg/ml, 20 µg/ml and 100 µg/ml of PMX-B, respectively. Furthermore, total TNFα production correlated significantly with increasing concentrations of PMX-B (R=0.53, P=0.007). We conclude that the use of PMX-B in in vitro studies as an LPS-neutralizing agent, or in the experimental treatment of endotoxic or septic shock can lead to erroneous interpretations of cytokine production by PBMC, and should be used cautiously in in vitro systems at high concentrations.

The Effect of Acupuncture on Proinflammatory Cytokine Production in Patients with Chronic Headache: A Preliminary Report

2003 ◽  
Vol 31 (06) ◽  
pp. 945-954 ◽  
Author(s):  
Hyun-Ja Jeong ◽  
Seung-Heon Hong ◽  
Yong-Che Nam ◽  
Hee-Sook Yang ◽  
Yeoung-Su Lyu ◽  
...  
Keyword(s):  
Chronic Headache ◽  
Mononuclear Cells ◽  
Cytokine Production ◽  
Acupuncture Treatment ◽  
Clinical Signs ◽  
Inhibitory Effect ◽  
Acute Stage ◽  
Control Group ◽  
Peripheral Blood Mononuclear ◽  
Tnf Α

Acupuncture has been widely used as a treatment for various conditions like headache and stroke, especially in Asian countries such as Korea and China. But few scientific investigations have been carried out. The aim of the present study is to investigate the effect of acupuncture on the production of inflammatory cytokines in patients with chronic headache (CH). Patients with CH were treated with acupuncture during the acute stage. Clinical signs of CH disappeared markedly after three months of treatment with acupuncture. Peripheral blood mononuclear cells obtained from a normal group and those from the patients with CH, before and after treatment with acupuncture, were cultured for 24 hours in the presence or absence of lipopolysaccharide (LPS). The amount of interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) in LPS culture supernatant was significantly increased in the patients with CH compared to the healthy control group (p < 0.05). But those cytokines came down toward the levels of the healthy group (p < 0.05) after treatment with acupuncture, although the levels still remained elevated. Plasma cytokine levels were analyzed to evaluate any change due to acupuncture treatment. There was little difference in the levels of IL-1β or IL-6 due to the treatment with acupuncture in the patients with CH, but significantly reduced plasma levels of TNF-α were observed. These data suggest that acupuncture treatment has an inhibitory effect on pro-inflammatory cytokine production in patients with CH.

scholarly journals Recognition of Streptococcus pneumoniae and Muramyl Dipeptide by NOD2 Results in Potent Induction of MMP-9, Which Can Be Controlled by Lipopolysaccharide Stimulation

2014 ◽  
Vol 82 (12) ◽  
pp. 4952-4958 ◽  
Author(s):  
Marloes Vissers ◽  
Yvonne Hartman ◽  
Laszlo Groh ◽  
Dirk J. de Jong ◽  
Marien I. de Jonge ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Mononuclear Cells ◽  
Muramyl Dipeptide ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Gram Negative Bacteria ◽  
Gram Positive ◽  
Potent Inducer ◽  
Gram Negative ◽  
Content Type ◽  
Gram Positive Bacteria

ABSTRACTMatrix metallopeptidase 9 (MMP-9) is a protease involved in the degradation of extracellular matrix collagen. Evidence suggests that MMP-9 is involved in pathogenesis duringStreptococcus pneumoniaeinfection. However, not much is known about the induction of MMP-9 and the regulatory processes involved. We show here that the Gram-positive bacteria used in this study induced large amounts of MMP-9, in contrast to the Gram-negative bacteria that were used. An important pathogen-associated molecular pattern (PAMP) for Gram-positive bacteria is muramyl dipeptide (MDP). MDP is a very potent inducer of MMP-9 and showed a dose-dependent MMP-9 induction. Experiments using peripheral blood mononuclear cells (PBMCs) from Crohn's disease patients with nonfunctional NOD2 showed that MMP-9 induction byStreptococcus pneumoniaeand MDP is NOD2 dependent. Increasing amounts of lipopolysaccharide (LPS), an important PAMP for Gram-negative bacteria, resulted in decreasing amounts of MMP-9. Moreover, the induction of MMP-9 by MDP could be counteracted by simultaneously adding LPS. The inhibition of MMP-9 expression by LPS was found to be regulated posttranscriptionally, independently of tissue inhibitor of metalloproteinase 1 (TIMP-1), an endogenous inhibitor of MMP-9. Collectively, these data show thatStreptococcus pneumoniaeis able to induce large amounts of MMP-9. These high MMP-9 levels are potentially involved inStreptococcus pneumoniaepathogenesis.

Endotoxin acts synergistically with C. difficile toxin B to increase IL-1 β production: A potential role for the intestinal biome in modifying the severity of C. difficile colitis

2021 ◽  
Author(s):  
P Htwe ◽  
H H Aung ◽  
B Kywe ◽  
P T Niang ◽  
T S Oo ◽  
...  
Keyword(s):  
Potential Role ◽  
Mononuclear Cells ◽  
Cytokine Production ◽  
Intestinal Microbiome ◽  
Peripheral Blood Mononuclear ◽  
Toxin B ◽  
E Coli ◽  
Tnf Α ◽  
Blood Mononuclear Cells ◽  
Stool Samples

Abstract. Background Inflammation is a crucial driver of host damage in patients with C. difficile colitis. We examined the potential for the intestinal microbiome to modify inflammation in patients with C. difficile colitis via the effects of gut-derived endotoxin on cytokine production. Methods Endotoxin from E. coli and P. aeruginosa as well as stool-derived endotoxin was tested for their ability to enhance IL-1β and TNFα- production by toxin B-stimulated peripheral blood mononuclear cells. Inflammasome and TLR-4 blocking studies were done to discern the importance of these pathways, while metagenomic studies were done to characterize predominant organisms from stool samples. Results Endotoxin significantly enhanced the ability of C. difficile toxin B to promote IL-1β production but not TNF- α. The magnitude of this effect varied by endotoxin type and was dependent on combined inflammasome and TLR-4 activation. Stool-derived endotoxin exhibited a similar synergistic effect on IL-1 β production with less synergy observed for stools that contained a high proportion of gamma-proteobacteria. Conclusions The ability of endotoxin to enhance IL-1 β production highlights a manner by which the microbiome can modify inflammation and severity of C. difficile disease. This information may be useful in devising new therapies for severe C. difficile colitis.

Psychological Stress and Cytokine Production in Multiple Sclerosis

2011 ◽  
Vol 15 (2) ◽  
pp. 226-233 ◽  
Author(s):  
Matthew Sorenson ◽  
Linda Janusek ◽  
Herbert Mathews
Keyword(s):  
Multiple Sclerosis ◽  
Psychological Stress ◽  
Perceived Stress ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Cytokine Production ◽  
Mood Disturbance ◽  
Peripheral Blood Mononuclear ◽  
Disease Symptoms ◽  
Tnf Α

Objective:Psychological variables such as perceived stress appear to play a role in symptom onset or disease exacerbation in multiple sclerosis (MS). The authors sought to determine if perceived stress is indeed associated with the expression of pro-inflammatory cytokines and disease symptoms in individuals with MS. To do so, the authors examined the relationships among disease symptomatology, perceived stress, and cytokine production from peripheral blood mononuclear cells in 42 outpatients with MS and 36 normative controls.Method:The authors drew peripheral blood from all subjects prior to the completion of a series of psychological instruments. The authors measured stress using the Perceived Stress scale and negative mood with the Profile of Mood States. Disease symptoms were measured using the Multiple Sclerosis Symptom Checklist. Cytokine production was induced separately by lipopolysaccharide and a combination of phytohemagglutinin and phorbol-12-myristate-13-acetate.Results:In MS subjects, the induced production of interleukin (IL)-6 and IL-10 positively correlated with psychological stress, mood disturbance, and disease symptomatology. In contrast, psychological stress in control subjects significantly correlated with level of tumor necrosis factor-alpha (TNF-α), and mood disturbance correlated with levels of TNF-α and interferon-gamma. As well, compared to controls, MS subjects exhibited a significant fourfold increase in the production of IL-12.Conclusion:There is, in those with MS, a pattern of IL-6 and IL-10 production that correlates significantly with perceived stress and disease symptomatology.

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