Human Intestinal Mast Cells Are Capable of Producing Different Cytokine Profiles: Role of IgE Receptor Cross-Linking and IL-4

Abstract

Mast cells are recognized as a new type of immunoregulatory cells capable of producing different cytokines. So far, little is known about the cytokine profile of mature human mast cells isolated from intestinal tissue and cultured in the presence of stem cell factor (SCF). We observed that these cells express the proinflammatory cytokines TNF-α, IL-1β, IL-6, IL-8, IL-16, and IL-18 without further stimulation. Both IgE-dependent and IgE-independent agonists (e.g., Gram-negative bacteria) enhanced expression of TNF-α. Another set of cytokines consisting of IL-3, IL-5, IL-9, and IL-13 was expressed following activation by IgE receptor cross-linking. If mast cells were cultured in the presence of IL-4 and SCF, the production and release of IL-3, IL-5, and IL-13 was increased up to 4-fold compared with mast cells cultured with SCF alone. By contrast, IL-6 expression was completely blocked in response to culture with IL-4. In summary, our data show that mature human mast cells produce proinflammatory cytokines that may be up-regulated following triggering with IgE-independent agonists such as bacteria, whereas activation by IgE receptor cross-linking results in the expression of Th2-type cytokines. IL-4 enhances the expression of Th2-type cytokines but does not affect or even down-regulates proinflammatory cytokines.

IL-4-Induced Priming of Human Intestinal Mast Cells for Enhanced Survival and Th2 Cytokine Generation Is Reversible and Associated with Increased Activity of ERK1/2 and c-Fos

“…We reported recently that IL-4, known to regulate B and T lymphocyte differentiation, strongly enhances growth and IgE-dependent mediator release of human intestinal mast cells (5). Moreover we found that IL-4 shifts human intestinal mast cell cytokine production toward a Th2 profile (6,7). Long-term challenge for 14 days with IL-4 and SCF compared with SCF alone enhanced the expression of IL-3, IL-5, and IL-13, which became detectable even without stimulation by Fc⑀RI cross-linking, whereas suppressing the production of IL-6.…”

“…The methods of mechanical and enzymatic tissue dispersion yielding single cell preparations containing 4 Ϯ 2% (mean Ϯ SD) mast cells have been described in detail elsewhere (4). After overnight incubation in culture medium (RPMI 1640 supplemented with 10% heat-inactivated FCS, 25 mM HEPES, 2 mM glutamine, 100 g/ml streptomycin, 100 g/ml gentamicin, 100 U/ml penicillin, and 0.5 g/ml amphotericin; all cell culture reagents were from Invitrogen Life Technologies), mast cells were enriched by positive selection of c-kit expressing cells using magnetic cell separation (MACS system; Miltenyi Biotec) and the mAb YB5.B8 (BD Pharmingen) as described (5,6). The fraction containing the c-kit-positive cells (mast cell purity 60 Ϯ 25%) was cultured at a density of 2 ϫ 10 5 mast cells per milliliter in medium supplemented with 50 ng/ml recombinant human SCF (Amgen) alone or in combination with 10 ng/ml IL-4 (Novartis Pharmaceuticals) and/or with 50 ng/ml IL-13 (PeproTech).…”

IL-18 contributes to the spontaneous development of atopic dermatitis-like inflammatory skin lesion independently of IgE/stat6 under specific pathogen-free conditions

“…Although the mechanism underlying AD is still elusive, activated T cells, basophils, and mast cells seem to play a crucial role in induction of AD. Allergen-dependent cross-linkage of FcR on basophils and mast cells activates them to produce Th2-related cytokines, such as IL-4, IL-13, and IL-5 and chemical mediators (3)(4)(5)(6)(7)(8), suggesting the importance of antigen (Ag)͞Ag-specific IgE in activation of basophils and mast cells (acquired type allergic response). However, we recently demonstrated the alternative, IgE-independent activation pathway (9,10).…”

Cytokine Production by Skin-Derived Mast Cells: Endogenous Proteases Are Responsible for Degradation of Cytokines

“…Intestinal mast cells produce IL-1␤, IL-3, IL-5, IL-6, IL-8, IL-9, IL-13, IL-16, IL-18, and TNF-␣ (18). Numerous studies have also examined cytokine production by the human mast cell leukemia cell line-1 (HMC-1) and by mast cells derived in vitro from cord blood or bone marrow progenitors (8, 19 -29).…”