scholarly journals Human Newborn Monocytes Demonstrate Distinct BCG-Induced Primary and Trained Innate Cytokine Production and Metabolic Activation In Vitro

2021 ◽  
Vol 12 ◽  
Author(s):  
Asimenia Angelidou ◽  
Joann Diray-Arce ◽  
Maria-Giulia Conti ◽  
Mihai G. Netea ◽  
Bastiaan A. Blok ◽  
...  
Keyword(s):  
Cytokine Production ◽  
Colorimetric Assay ◽  
Age Groups ◽  
Metabolic Activation ◽  
Human Monocyte ◽  
Autologous Serum ◽  
Live Attenuated Vaccine ◽  
Cytokine Responses ◽  
The Impact

BackgroundNewborns exhibit distinct immune responses and are at high risk of infection. Neonatal immunization with BCG, the live attenuated vaccine against tuberculosis (TB), is associated with broad protection against a range of unrelated pathogens, possibly reflecting vaccine-induced training of innate immune cells (“innate memory”). However, little is known regarding the impact of age on BCG-induced innate responses.ObjectiveEstablish an age-specific human monocyte in vitro training platform to characterize and compare BCG-induced primary and memory cytokine responses and immunometabolic shifts.Design/MethodsHuman neonatal and adult CD33-selected monocytes were stimulated for 24h with RPMI (control) or BCG (Danish strain) in 10% autologous serum, washed and cultured for 5 additional days, prior to re-stimulation with the TLR4 agonist LPS for another 24h. Supernatants were collected at Day 1 (D1) to measure primary innate responses and at Day 7 (D7) to assess memory innate responses by ELISA and multiplex cytokine and chemokine assays. Lactate, a signature metabolite increased during trained immunity, was measured by colorimetric assay.ResultsCytokine production by human monocytes differed significantly by age at D1 (primary, BCG 1:750 and 1:100 vol/vol, p<0.0001) and D7 (innate memory response, BCG 1:100 vol/vol, p<0.05). Compared to RPMI control, newborn monocytes demonstrated greater TNF (1:100, 1:10 vol/vol, p<0.01) and IL-12p40 (1:100 vol/vol, p<0.05) production than adult monocytes (1:100, p<0.05). At D7, while BCG-trained adult monocytes, as previously reported, demonstrated enhanced LPS-induced TNF production, BCG-trained newborn monocytes demonstrated tolerization, as evidenced by significantly diminished subsequent LPS-induced TNF (RPMI vs. BCG 1:10, p <0.01), IL-10 and CCL5 production (p<0.05). With the exception of IL-1RA production by newborn monocytes, BCG-induced monocyte production of D1 cytokines/chemokines was inversely correlated with D7 LPS-induced TNF in both age groups (p<0.0001). Compared to BCG-trained adult monocytes, newborn monocytes demonstrated markedly impaired BCG-induced production of lactate, a metabolite implicated in immune training in adults.ConclusionsBCG-induced human monocyte primary- and memory-innate cytokine responses were age-dependent and accompanied by distinct immunometabolic shifts that impact both glycolysis and training. Our results suggest that immune ontogeny may shape innate responses to live attenuated vaccines, suggesting age-specific approaches to leverage innate training for broad protection against infection.

scholarly journals P068 The combination of JAK1 and TPL2 or IRAK4 inhibitors is more effective than single agents in reducing TLR-mediated cytokine responses in human monocyte populations

2021 ◽  
Vol 15 (Supplement_1) ◽  
pp. S173-S173
Author(s):  
E Hornsby ◽  
A N Yadon ◽  
A Clarke ◽  
E Grant ◽  
J O Lindsay ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Cytokine Production ◽  
Inflammatory Responses ◽  
Human Monocyte ◽  
Signalling Pathways ◽  
Tnf Alpha ◽  
Monocyte Subsets ◽  
Cytokine Responses ◽  
Erk Phosphorylation ◽  
The Impact

Abstract Background Dysregulated Toll-like receptor (TLR)-mediated responses in intestinal monocyte-derived cells contribute to pathology in inflammatory bowel disease (IBD). JAK inhibitors regulate the production of inflammatory mediators and are an emerging therapy in IBD. Recent data suggest that combination therapies may be more effective than single agents, and therapies targeting aspects of TLR signalling pathways are being developed. We assessed the impact of inhibitors of IRAK4, TPL2 and JAK1 (IRAK4i, TPL2i and JAK1i, respectively) either alone or in combination on TLR-mediated cytokine responses and associated signalling pathways in classical (CD14+CD16-), intermediate (CD14+CD16+) and non-classical (CD14lowCD16+) human blood monocyte subsets. Methods Peripheral Blood Mononuclear Cells (PBMCs) or CD14+ monocytes from healthy volunteers were pre-incubated with TPL2i, IRAK4i or JAK1i individually or with JAK1i in combination with TPL2i or IRAK4i for 1 hour at 37°C. Cells were then stimulated at 37°C with agonists of TLR4 (E.coli lipopolysaccharide; LPS) or TLR2 (Pam3CYSK4). Intra-cellular staining and flow cytometry were used to measure phosphorylation of signalling molecules (NF-kappaB p65, p38 MAPK and ERK) or cytokines (TNF-alpha and IL-1-beta) in the three monocyte subsets after stimulation for 15 minutes or 3 hours respectively. Results Pre-incubation of PBMCs or CD14+ monocytes with IRAK4i, TPL2i or JAK1i individually led to a significant dose-dependent reduction in TLR-stimulated cytokine production (the frequency of TNF-alpha+ cells and the per cell level of IL-1-beta) within the classical, intermediate and non-classical monocyte subsets. The combination of JAK1i with IRAK4i or TPL2i had an additive effect. TPL2i reduced LPS-stimulated ERK phosphorylation, but p65 and p38 phosphorylation in response to LPS was not affected by any of the inhibitors. Conclusion Small molecule inhibitors of TPL2, IRAK4 and JAK1 dampen TLR4- and TLR2- mediated inflammatory responses in human monocyte subsets, by affecting pathways independent of p65 or p38. This is due to a cell intrinsic effect on CD14+ monocytes. TPL2 inhibition may in part act to dampen cytokine production in monocytes through the reduction of ERK phosphorylation. Combining JAK1i with IRAK4i or TPL2i may be more effective in reducing inflammatory responses than single agents. Further work is required to elucidate the mechanisms by which IRAK4i and JAK1i dampen TLR-mediated inflammation in monocyte subsets.

scholarly journals Keratinocytes Regulate the Threshold of Inflammation by Inhibiting T Cell Effector Functions

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1606
Author(s):  
Peter Seiringer ◽  
Stefanie Eyerich ◽  
Kilian Eyerich ◽  
Daniela Dittlein ◽  
Anna Caroline Pilz ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
T Cell ◽  
Cytokine Production ◽  
Nickel Sulfate ◽  
Human Keratinocytes ◽  
T Cell Proliferation ◽  
Effector Functions ◽  
The Impact ◽  
Cell Effector

Whilst the importance of keratinocytes as a first-line defense has been widely investigated, little is known about their interactions with non-resident immune cells. In this study, the impact of human keratinocytes on T cell effector functions was analyzed in an antigen-specific in vitro model of allergic contact dermatitis (ACD) to nickel sulfate. Keratinocytes partially inhibited T cell proliferation and cytokine production. This effect was dependent on the keratinocyte/T cell ratio and was partially reversible by increasing the number of autologous dendritic cells. The inhibition of T cell proliferation by keratinocytes was independent of the T cell subtype and antigen presentation by different professional antigen-presenting cells. Autologous and heterologous keratinocytes showed comparable effects, while the fixation of keratinocytes with paraformaldehyde abrogated the immunosuppressive effect. The separation of keratinocytes and T cells by a transwell chamber, as well as a cell-free keratinocyte supernatant, inhibited T cell effector functions to the same amount as directly co-cultured keratinocytes, thus proving that soluble factor/s account for the observed suppressive effects. In conclusion, keratinocytes critically control the threshold of inflammatory processes in the skin by inhibiting T cell proliferation and cytokine production.

scholarly journals Human monocyte isolation methods influence cytokine production from in vitro generated dendritic cells

Immunology ◽  
2005 ◽  
Vol 114 (2) ◽  
pp. 204-212 ◽  
Author(s):  
Eyad Elkord ◽  
Paul E. Williams ◽  
Howard Kynaston ◽  
Anthony W. Rowbottom
Keyword(s):  
Dendritic Cells ◽  
Cytokine Production ◽  
Human Monocyte ◽  
Isolation Methods ◽  
Monocyte Isolation

scholarly journals THE IMPACT OF ELECTROMAGNETIC RADIATION OF MOBILE PHONE FOR LYMPHOCYTE ACTIVATION IN VITRO

2019 ◽  
Vol 96 (10) ◽  
pp. 965-970 ◽  
Author(s):  
Maria S. Blyakher ◽  
E. A. Tulskaya ◽  
I. V. Kapustin ◽  
I. M. Fedorova ◽  
T. K. Lopatina ◽  
...  
Keyword(s):  
Electromagnetic Radiation ◽  
Mobile Phone ◽  
Blood Cells ◽  
Cytokine Production ◽  
Venous Blood ◽  
Peripheral Blood Lymphocytes ◽  
Mean Values ◽  
Healthy Donors ◽  
The Impact

The character of the influence of the electromagnetic radiation (EMR) of mobile phone on the activation of lymphocytes in vitro was investigated. This is important, since modern human is exposed to a complex combination of electric and magnetic fields (EMF) of different frequencies. The object of the study were whole venous blood and lymphocytes isolated from 21 adult donors (aged of from 20 to 55 years) - 10 were healthy donors and 11 were healthy persons 7 days after their vaccination with meningococcal polysaccharide vaccine. In the study the influence of phone’s EMR on the functional activity of peripheral blood lymphocytes was determined by the flow cytometry method with the use of monoclonal antibodies of Beckman Coulter company (by the identification and calculation the number of basic and activated lymphocyte subpopulations). The changes of cytokines production by blood cells exposed to mobile phone electromagnetic radiation were determined in supernatants by measuring their concentration using EIA kits produced by JSC “Vector-Best” (Russia) and LLC “cytokine” (Russia). The results of the study of the effects of electromagnetic radiation of mobile phone on blood cells revealed changes in the percentage of lymphocytes carrying the early activation marker CD69 significantly to be more frequently and were observed with greater intensity in the group of donors which were vaccinated compared to healthy donors. Under the influence of phone’s EMR mean values of cytokine production determined in the supernatants samples did not changed in both groups, but in the group of healthy donors mean values of cytokines production were 1,5 - 2 times higher than in the group of persons following immunization. The increase or decrease in cytokine production under the influence of phone’s EMR occurred regardless of the initial level of its production in the surveyed donor. The changes of the cytokine production (IFNγ, TNFα, IL-6 and IL-8) by blood cells under the influence of phone’s EMR happen individually; this should be considered when deciding on the presence or absence of phone’s EMR impact on the status of lymphocytes.

Effect of oxidant challenge on contractile function of the aging rat diaphragm

1997 ◽  
Vol 272 (2) ◽  
pp. E201-E207 ◽  
Author(s):  
J. M. Lawler ◽  
C. C. Cline ◽  
Z. Hu ◽  
J. R. Coast
Keyword(s):  
Relaxation Time ◽  
Xanthine Oxidase ◽  
Contractile Function ◽  
Age Groups ◽  
Low Frequency ◽  
Fiber Bundles ◽  
Stimulation Protocol ◽  
Time To Peak ◽  
The Impact

Although controversial, growing evidence indicates that reactive oxygen species (ROS) alter contractions of skeletal muscle, including the diaphragm. However, the impact of ROS on contractility of the aging diaphragm is unknown. The xanthine oxidase (0.01 U/ml) system was used as an ROS generator, imposing an oxidant challenge. Contractile function [twitch tension; twitch time to peak tension; twitch one-half relaxation time; tension at 10 and 20 Hz; maximal tetanic tension (Po) at 100 Hz] of costal diaphragm fiber bundles from young (4 mo) and old (25 mo) Fischer 344 rats was examined in vitro before and after treatment with control Krebs solution [young control (YC) and old control (OC)], or with xanthine oxidase (XO; 0.01 U/ml) plus hypoxanthine (0.29 mg/ml) substrate [young XO treated (YXO) and old XO treated (OXO)]. Contractile function of fiber bundles was reassessed after oxidant challenge in an unfatigued state (Post-u) or 10 min after a fatiguing stimulation protocol (Post-f). Oxidant challenge in the unfatigued fiber bundles increased twitch tension and tension at 10 and 20 Hz in YXO, but not OXO, without increasing Po. Conversely, XO significantly depressed fatigued diaphragm twitch and low-frequency tension in both OXO and YXO, compared with controls. Po was depressed Post-f in OXO but not YXO. Oxidant challenge also increased twitch one-half relaxation time of the fatigued diaphragm in both age groups. Furthermore, fiber bundles from old rats suffered greater fatigue during the stimulation protocol. We conclude that the response to oxidant challenge and increased contractile demand is impaired in the aging diaphragm.

scholarly journals An in vitro model for dengue virus infection that exhibits human monocyte infection, multiple cytokine production and dexamethasone immunomodulation

2007 ◽  
Vol 102 (8) ◽  
pp. 983-990 ◽  
Author(s):  
Sônia Regina Nogueira Ignácio Reis ◽  
André Luiz Franco Sampaio ◽  
Maria das Graças Muller Henriques ◽  
Mariana Gandini ◽  
Elzinandes Leal Azeredo ◽  
...  
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
In Vitro Model ◽  
Cytokine Production ◽  
Human Monocyte ◽  
Dengue Virus Infection ◽  
Vitro Model ◽  
Monocyte Infection

Effect of recombinant Panton–Valentine leukocidin in vitro on apoptosis and cytokine production of human alveolar macrophages

2010 ◽  
Vol 56 (3) ◽  
pp. 229-235 ◽  
Author(s):  
Benquan Wu ◽  
Wenxian Zhang ◽  
Jing Huang ◽  
Hui Liu ◽  
Tiantuo Zhang
Keyword(s):  
Alveolar Macrophages ◽  
Cytokine Production ◽  
Lavage Fluid ◽  
Effector Cells ◽  
Tnf Α ◽  
Human Alveolar Macrophages ◽  
The Impact ◽  
Panton Valentine Leukocidin ◽  
Valentine Leukocidin

Panton–Valentine leukocidin (PVL) is associated with rare cases of necrotizing pneumonia that occur in otherwise healthy individuals. Human alveolar macrophages (HAMs) are major effector cells in host defense against infections. However, the impact of PVL on HAMs is uncertain. We evaluated the role of PVL in cytotoxicity and production of inflammatory cytokines secreted by HAMs. HAMs were purified from bronchoalveolar lavage fluid. Recombinant PVL (rPVL) was used in the study to interfere with HAM apoptosis and cytokine production in vitro. Hoechst 33342 fluorescence staining, transmission electron microscopy examination, and flow cytometry indicated that rPVL (10 nmol/L) treatment resulted in HAMs with markedly apoptotic characteristics, and HAMs treated with rPVL at 100 nmol/L showed clear indication of necrosis. A treatment of rPVL at 10 nmol/L elicited the secretion of IL-10 by HAMs relative to untreated control cells, but there was a slight decrease in the constitutive secretion of tumor necrosis factor (TNF)-α. Our results indicate that PVL-treated samples decreased HAM viability, leading to apoptosis at low concentrations and necrosis at high concentrations. In addition, PVL-treated cells released increased amounts of IL-10 and decreased amounts of TNF-α under apoptosis-inducing concentrations. Therefore, we speculated that PVL could play a negative role in HAM function at lower concentrations.

scholarly journals Lysozyme enhances monocyte-mediated tumoricidal activity: a potential amplifying mechanism of tumor killing

Blood ◽  
1981 ◽  
Vol 58 (5) ◽  
pp. 994-999 ◽  
Author(s):  
P LeMarbre ◽  
JJ Rinehart ◽  
NE Kay ◽  
R Vesella ◽  
HS Jacob
Keyword(s):  
Tumor Cell ◽  
Metabolic Activation ◽  
Human Monocyte ◽  
Cell Interaction ◽  
Competitive Inhibitor ◽  
Mononuclear Phagocyte ◽  
Mononuclear Phagocytes ◽  
Tumoricidal Activity

The mononuclear phagocyte is well established as an in vitro cytotoxic effector cell for certain human tumors. The mechanism(s) for this action remains unclear. Increased levels of lysozyme, a cationic enzyme synthesized in large amounts by mononuclear phagocytes, are associated with increased resistance to transplantable animal tumors. In this study, we provide evidence that human lysozyme, isolated from the urine of leukemic patients, has marked potentiating effects on human monocyte- tumor-cell cytocidal activity. In addition, lysozyme-exposed monocytes incorporate increased quantities of leucine, suggesting that monocytes are capable of amplifying their own metabolic activation by secreting an endogenous constituent. Tri-N-acetyl-glucosamine, a competitive inhibitor for the active site of lysozyme, inhibits cytocidal activity. Conversely, protamine, an extraneous albeit similarly positively charged molecule, increases monocyte-mediated tumor cytotoxicity; this protamine effect is negated by heparin. We conclude that lysozyme, at least partially by its positive charge, is capable of enhancing in vitro monocyte tumor cell cytotoxicity; its in vivo secretion may potentiate monocyte-tumor-cell interaction.

P4467PCSK9 and Lp(a) levels of children born after assisted reproduction technologies: results from a pilot study

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
I Koutagiar ◽  
C Vlachopoulos ◽  
D Terentes-Printzios ◽  
I Kosteria ◽  
S Sakka ◽  
...  
Keyword(s):  
Cardiovascular Risk ◽  
Assisted Reproduction ◽  
Age Groups ◽  
Blood Lipid ◽  
Blood Lipid Profile ◽  
Vitro Fertilization ◽  
Assisted Reproduction Technologies ◽  
Patient Series ◽  
The Impact

Abstract Background Since the introduction of Assisted Reproduction Technologies (ART) in clinical practice several studies have addressed concerns regarding the long-term health of the offspring and have revealed indications of an adverse cardiovascular/cardiometabolic outcome. Proprotein convertase subtilisin/kexin type 9 (PCSK9) and lipoprotein (a) (Lp[a]) levels have been associated with cardiovascular risk. Purpose To investigate PCSK9 and Lp(a) levels of children born after ART compared with naturally conceived (NC) controls. Methods In this case-control study, 73 sex- and age-matched children (mean age 98±35 months) of ART (intracytoplasmic sperm injection [ICSI]: n=33, classic in vitro fertilization [IVF]: n=40) and 73 NC children were assessed. Blood lipid profile, including PCSK9 and Lp(a) levels, was measured. Children were grouped according to age (<8 years, 8–10 years, ≥10 years). Results In the univariate model of the overall population, circulating PCSK9 levels were related to total cholesterol (r=0.186, P=0.025), LDL-C (r=0.180, P=0.029) and SBP (r=0.199, P=0.021). Similarly, circulating Lp(a) levels were related to age (r=0.269, P=0.001), apoB (r=0.214, P=0.01), birth weight (r=−0.183, P=0.037), height (r=0.263, P=0.001), waist-to-hip ratio (r=−0.350, P<0.001), HOMA-IR (r=0.319, P<0.001), insulin (r=0.316, P<0.001), and hsCRP (ρ=0.241, P=0.018). No significant differences were observed regarding lipid biomarkers between ART and NC children. (p=0.515 for PCSK9 and p=0.277 for Lp(a) values). It is noteworthy that a significant interaction was found between age groups and conception method (P<0.001) showing that PCSK9 levels increase with age in ART children, while they decline with age in NC offspring (Figure 1). IVF children showed higher levels of adjusted mean Lp(a) than ICSI (13.5 vs. 6.8 mg/dl, P=0.010) and NC children (12.3 vs. 8.3 mg/dl, P=0.048). Mean LogPCSK9 concentrations with the st Conclusions PCSK9 and Lp(a) levels did not differ between ART and NC children. Nonetheless, PCSK9 levels increase with age in ART children indicating a gradual deterioration of lipidemic profile that could lead to increased cardiovascular risk. Moreover, our results imply that ART method may be of importance given that classic IVF is associated with higher levels of Lp(a). The impact of the method of conception on PCSK9 and Lp(a) values should be validated in larger patient series.

A pilot study on the effect of Lactobacillus casei Zhang on intestinal microbiota parameters in Chinese subjects of different age

2014 ◽  
Vol 5 (3) ◽  
pp. 295-304 ◽  
Author(s):  
L.Y. Kwok ◽  
L. Wang ◽  
J. Zhang ◽  
Z. Guo ◽  
H. Zhang
Keyword(s):  
Lactobacillus Casei ◽  
Age Groups ◽  
Short Chain Fatty Acids ◽  
Fermented Milk ◽  
In Vitro Tests ◽  
Probiotic Properties ◽  
Lactobacillus Casei Zhang ◽  
The Impact ◽  
Chinese Subjects

Ageing of the population is an imminent global problem. Lactobacillus casei Zhang (LcZ) was isolated from Inner Mongolian fermented milk, koumiss. LcZ possesses numerous probiotic properties in in vitro tests and in animal models. However, it has never been tested in any human trial. In the current study, the impact of oral consumption of LcZ on different age groups was tested. Chinese subjects, including 10 young, 7 middle-aged and 7 elderly volunteers (with mean age of 24.3, 47.6 and 64.7, respectively), were recruited. Each subject took 10.6 log10 cfu LcZ daily for a continuous period of 28 days. Several parameters, including the amounts of LcZ and four selected groups of bacteria, change of bacterial diversity, short chain fatty acids (SCFA) and total bile acids (TBA), were monitored in faecal samples collected from the subjects before starting, during and after stopping oral LcZ consumption. The consumption of LcZ exhibited beneficial effects to the subjects by modulating faecal microbiota in a temporal manner with a prolonged elevation of SCFA and reduction of TBA. The potentially harmful Pseudomonas and Acinetobacter genera were suppressed by the probiotic administration. Furthermore, a moderately divergent response was observed in the indigenous gut populations of Bifidobacterium and Bacteroides fragilis group in different age subjects. Taken together, the current study has provided proof on the positive effect of probiotic consumption and crucial insights into the design and application of probiotic-based products to users of different age segments.

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