Identification, purification, and characterization of a mast cell-associated cytolytic factor related to tumor necrosis factor.

Abstract

The role of mast cells and mast-cell-derived factors in natural cytotoxic reactions was investigated. Cultured and freshly isolated murine mast cells are shown to be cytotoxic to WEHI-164 and YAC-1 targets in 18-hr viability assays but not in 4-hr assays. Here, we describe a cytotoxic factor in murine mast cells that is immunologically related to tumor necrosis factor (TNF). This TNF-like factor lyses WEHI-164 cells with a slow time course requiring 16-20 hr for the lytic reaction to complete. Antibodies specific for human and murine TNF and human lymphotoxin partially block mast cell lysis of WEHI-164 cells. These antibodies react on immunoblots with one major mast cell protein band of 50 kDa. Immunoblot analysis shows this factor in cloned and uncloned cultured mouse mast cells and in mature "connective tissue-type" mast cells freshly purified from rat or mouse peritoneal cavities. The amount of this factor is greatly enhanced in cells that have been stimulated with a combination of phorbol ester/concanavalin A or bacterial lipopolysaccharide. Subcellular fractionation analysis of mast cells with Percoll gradients reveals two pools of TNF-related cytotoxic activity that are associated with free cytosolic material and granule fractions. In contrast to cytotoxic T lymphocytes and natural killer cells, granuleenriched fractions of mast cells do not contain any hemolytic activity. The localization of the TNF-like molecule in mast cell granules may play a strategical role in the rapid delivery of this mediator to the target cell membrane following cell surface stimulation and degranulation.

Augmentation of reverse arthus reaction by mast cells in mice.

“…Antihistamines will decrease plasma leakage during immune complex reactions (34). It has recently been reported that mast cells upon stimulation with a number of agents, including IgE/Ag, calcium ionophore, and PMA, will cause the induction of various cytokines, such as IL-1, IL-3, IL-4, IL-6, GM-CSF, and TNFa (35)(36)(37) (42).…”

Tumour necrosis factor: a cytokine with multiple biological activities

“…In fact, although the major source of this cytokine is macrophages (Beutler & Cerami, 1987;Aggarwel et al, 1985), the molecule can also be released by other cell types. These include monocytes stimulated by gamma interferon (Beutler & Cerami, 1987), T lymphocytes and T cell lines following stimulation with phorbol esters and anti-CD3 antibodies in different combinations (Granger & Williams, 1968;Cutri et al, 1987;Scheurich et al, 1987;Sung et al, 1988a;Turner et al, 1987), B lymphocytes (Williamson et al, 1983;Sung et al, 1988b), large granular lymphocytes (Ostensen et al, 1987;Peters et al, 1986) and mast cells (Young et al, 1987).…”

Resident mast cells are important for eotaxin-induced eosinophil accumulation in vivo

“…Bone marrow-derived MC line was prepared from BALB/c mice as reported [14,15]. Cells were grown in DMEM supplemented with 10% FCS, antibiotics, and 0.1% mercaptoethanol and passaged every other day to keep their concentration in the range of 0.3 to 0.5 ϫ 10 6 /mL.…”