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Human exposure to engineered nanomaterials (ENMs) is inevitable due to the plethora of applications they are being manufactured for and integrated within. ENMs demonstrate plentiful advantages in terms of industrial approaches, as well as from a consumer perspective. However, despite such positives, doubts remain over the human health implications...
Introduction: Eosinophils have been long implicated in anti-parasite immunity and allergic diseases and, more recently, in regulating adipose tissue homeostasis. The metabolic processes that govern eosinophils, particularly upon activation, are unknown. Methods: Peripheral blood eosinophils were isolated for analysis of metabolic processes using...
Background and Aims: Ascites and spontaneous bacterial peritonitis (SBP) are frequent complications of liver cirrhosis. In spite of the clinical impact, knowledge about ascites as an immune cell compartment in liver disease is limited. Therefore, we analyzed NK cells in blood, ascites, and liver. Methods: Mononuclear cells from blood, ascites, and...
Autoimmune hepatitis (AIH) is an immune-mediated disease with no curative treatment. Regulatory T cell (Treg) therapy is potentially curative in AIH given the critical role of Tregs in preventing autoimmunity. To work effectively, adoptively transferred Tregs must migrate to and survive within the inflamed liver. We conducted a proof-of-concept stu...
Metabolic pathways that regulate T-cell function show promise as therapeutic targets in diverse diseases. Here, we show that at rest cultured human effector memory and central memory CD4+ T-cells have elevated levels of glycolysis and oxidative phosphorylation (OXPHOS), in comparison to naïve T-cells. Despite having low resting metabolic rates, nai...
Linking immunometabolic adaptation to T-cell function provides insight for the development of new therapeutic approaches in multiple disease settings. T-cell activation and downstream effector functions of CD4+ and CD8+ T-cells are controlled by the strength of interaction between the T-cell receptor (TCR) and peptides presented by human leukocyte...
A biomarker is an accurately and reproducibly quantifiable biological characteristic that provides an objective measure of health status or disease. Benefits of biomarkers include identification of therapeutic targets, monitoring of clinical interventions, and development of personalized (or precision) medicine. Challenges to the use of biomarkers...
Leukocytes respond rapidly to pathogenic and other insults with responses ranging from cytokine production through to migration and phagocytosis. These are bioenergetically expensive and increased glycolytic flux provides ATP rapidly to support these essential functions. However, much of this work is from animal studies. To better understand the re...
Questions and Answers
Question & Answers (5)
Does anyone have any experience activating human naive CD4 T cells (0.25 x10^6)? I am trying to activate them with immobilized anti-CD3 (2ug/mL) with soluble CD28 (20ug/mL) with RPMI/5% FBS but it doesn't appear to be working. I have tried both free anti-CD3 and 28 also and measured IFNg as output with no luck. Can anyone please advise, cheers?
I am looking to stimulate neutrophils with LPS (tried 10 ng/mL, 0.1 ug/mL and 1 ug/mL) with 0.25 x 10^6 cells per well (200 ul of a 96 well plate) in RPMI + Glutamax. After performing the IL-8 ELISA there was no difference between untreated and LPS treated for all concentrations. Culture times were 6, 9 and 24h. The ELISA was functioning properly but no IL-8 detected at all. The only thing I can think of is I did not include FCS in the culture media however the cells looked alive. Any advice greatly appreciated.
I can get the positive control to work but nothing else does.
Neutrophils 2 x 10^5 per well
Add 50ng/mL GM-CSF for 15 min
Add 10ng/mL LPS for 1 hr
Spin plate, remove super 50uL with 50uL TMB for 10 min
Basically the positive (lysed with water) works fine, the neutrophils appear activated but are not degranulating as I have no difference between control and GM-CSF/LPS treated? Any advice is greatly appreciated