Dual Phase Priming by IL-3 for Leukotriene C4 Generation in Human Basophils: Difference in Characteristics Between Acute and Late Priming Effects

Abstract

Previous studies have suggested that enhancement of mediator release from human basophils by IL-3 occurs in at least two phases, and the current studies further characterize the signaling changes that accompany these two phases of the basophil in response to IL-3. The test stimulus for these studies was anaphylatoxin split product of C component (C5a), which does not induce leukotriene C4 release without prior IL-3 treatment. Functionally, IL-3 priming occurs after 5 min, disappears by 2 h, and returns by 18 h. In contrast, the kinetics of cytosolic phospholipase A2 (cPLA2) and extracellular signal-regulated kinase (ERK1/2) phosphorylation, induced by IL-3, do not show the second rise by 18 h. The kinetics of cPLA2 and ERK1/2 phosphorylation following stimulation with C5a are the same for cells that were not treated with IL-3 as for those treated for 18 h, i.e., a lag in phosphorylation of cPLA2 and ERK1/2 lasting 30 s before its eventual rise. Previous studies showed that a 5-min treatment with IL-3 induced little change in the C5a-induced cytosolic calcium response, while 24 h of treatment resulted in a marked and sustained cytosolic calcium elevation during the C5a-induced response. The first phase of the IL-3 priming effect (5–15 min of treatment) was unaffected by cycloheximide, while the second phase (18 h) was inhibited. These data suggest that early IL-3 priming results from preconditioning cPLA2, i.e., causing its phosphorylation, while late priming results from a qualitative change in the cytosolic calcium response.

Granzyme B, a novel mediator of allergic inflammation: its induction and release in blood basophils and human asthma

“…We chose 4 growth factors, IL-3, IL-5, GM-CSF, and NGF, which all acutely prime basophils with similar efficacy, 10,25,26 as well as cytokines thought to enhance cytotoxic functions of lymphocytes (IL-2, interferon-␣, and interferon-␥). The efficacy of IL-3 to induce GzmB was rather unique since only weak effects were observed in 31,32 and the induction of GzmB expression shows an identical pattern and order of efficacy of the cytokines (IL-3 Ͼ Ͼ Ͼ NGF Ͼ IL-5 Ͼ GM-CSF), while IL-3 and NGF were similarly effective in enhancing C5a-induced degranulation. A similar pattern of responses was found using different doses of IL-3.…”

“…The most efficient cytokine inducing GzmB together with other phenotypic and functional alterations is IL-3. [30][31][32] An identical concentration range of IL-3 induces GzmB expression and the persistent facilitation of LTC 4 formation. When the effect of different cytokines was compared, we observed an identical order of efficacy.…”

“…10,11,[13][14][15]25 Furthermore, IL-3 has the longest duration of action and can also induce phenotypic alterations of mature basophils. 12,15,[30][31][32] From the Institute of Immunology, Inselspital, University of Bern, Switzerland; the Department of Pneumology, University Medical Clinic, Rostock, Germany; and Sanquin Research, Amsterdam, the Netherlands. For personal use only.…”

C5a-induced in vitro basophil activation in patients with chronic urticaria: a pilot study

“…Nevertheless, the results of their study may have been influenced by technical problems, as basophile histamine release after C5a stimulation was very low in the healthy controls, who released only around 4% of the histamine content after C5a stimulation. In contrast, an earlier study by Miura and MacGlashan [17], for example, and a recent study by Marsland et al [18] have shown that basophils of normal donors release about 40% of their histamine content after C5a stimulation. Calcineurin inhibitors were able to inhibit the in vitro release of histamine when basophils were stimulated with urticaria sera but not when stimulated with C5a [18].…”

IgE-Dependent and IgE-Independent Stimulation of Human Basophils Increases the Presence of Immature FcεRIα by Reversing Degradative Pathways

“…We found that MG-132 and other proteasome inhibitors induced significant apoptosis of basophils cultured for only 18–24 h. This result led to the conclusion that results using agents that altered apoptosis would be difficult to interpret. Although 18 h of treatment of basophils with cycloheximide does not appear to cause changes in viability [29], incubation with protein synthesis or transcriptional inhibition reverse the effects of IL-3 [29], and IL-3 promotes survival in basophils [28,30]. In light of this, the actinomycin D and cycloheximide data should be interpreted cautiously.…”