The effects of recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3 on the secretory capacity of human blood eosinophils

Abstract

SUMMARYThe effects of recombinant human GM-CSF and interleukin-3 (IL-3) on human blood eosinophil survival, activation, and secretion were studied. Purified normal density eosinophils from patients with the idiopathic hypereosinophilic syndrome (HES) survived in culture for 7 days (50% viable) in the presence of 50 nM GM-CSF or 50 nm IL-3. Neutrophils did not survive after 4 days. No eosinophils survived in the absence of GM-CSF or IL-3. In two out of five patients studied, the cultured eosinophils became elongated with numerous processes. In all five patients the cells became adherent, but there were no morphological signs of degranulation. Both GM-CSF and IL-3 activated eosinophils, transforming the storage form of eosinophil cationic protein (ECP) into the secreted form. The proportion of activated cells increased from less than 20% to over 50% after 4 days in culture. However, GM-CSF and IL-3 did not induce secretion on their own. On the other hand, when GM-CSF/IL-3-activated eosinophils were exposed to known secretory stimuli, there was a six-fold increase in the amount of ECP released when the cells were stimulated with sepharose coated with C3b, and a two-fold increase when they were stimulated with sepharose-activated whole autologous serum. Eosinophils from patients taking steroids were unable to secrete their granule contents, even though they became activated by GM-CSF and IL-3. A novel finding was that sepharose-activated whole serum was an extremely potent secretory signal for ECP, releasing up to 50% of the total ECP content. These studies showed that GM-CSF and IL-3 prime eosinophil effector function by initiating granule solubilization which is the first step in the secretory event, without affecting the subsequent extracellular release of granule proteins.

miR-143 inhibits interleukin-13-induced inflammatory cytokine and mucus production in nasal epithelial cells from allergic rhinitis patients by targeting IL13Rα1

“…GM-CSF can also stimulate hematopoietic stem cells to differentiate into granulocytes and monocytes and can promote the survival and activation of airway inflammatory cells [20,21]. As a C-C chemotactic protein, eotaxin can recruit immune cells, such as eosinophils, Th2 cells, basophils, and mast cells, to inflammatory sites in allergic diseases by specifically binding to the chemokine receptor CCR3 [7,22].…”

Alternaria Fungus Induces the Production of GM-CSF, Interleukin-6 and Interleukin-8 and Calcium Signaling in Human Airway Epithelium through Protease-Activated Receptor 2

“…GM-CSF is known as a pleiotropic and proinflammatory cytokine and is a major survival and activating factor for hematopoietic cells including mature macrophages, eosinophils and neutrophils [30]. The respiratory epithelium produces significant amounts of GM-CSF, and infiltrating leukocytes can be induced to synthesize GM-CSF as an autocrine growth factor by inflammatory and chemotactic stimuli.…”

Inhibition of IL-32 and TSLP production through the attenuation of caspase-1 activation in an animal model of allergic rhinitis by Naju Jjok (Polygonum tinctorium)

“…Eol-1 cells (3x10 6 ) were treated with NJJ (1, 10, or 100 mg/ml) for 1 h prior to stimulation with GM-CSF and incubated for 4 h. Total RNA was isolated from the cells and nasal mucosal tissue according to the manufacturer's specifications using an easy-BLUE™ RNA extraction kit (iNtRON Biotech, Sungnam, Korea). The concentration of total eluted RNA was determined by spectrophotometry.…”

“…Caspase-1 assay. The Eol-1 cells (3x10 6 ) were treated with NJJ (1, 10 or 100 µg/ml) for 1 h prior to stimulation with GM-CSF and incubated for 2 h. Caspase-1 assay was used to analyze the nasal mucosal tissue and cell extracts. Caspase-1 activity was measured according to the manufacturer's specifications using the caspase assay kit (R&D Systems Inc.).…”

“…Activated endothelial cells express intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on their cell surfaces (5). GM-CSF is an important activating factor for eosinophils and neutrophils, and is known as a pleiotropic and pro-inflammatory cytokine (6). GM-CSF induced the expression of IL-32 in eosinophils (7).…”