鼻アレルギー患者鼻粘膜上皮細胞培養上清中の好塩基性細胞および好酸球分化増殖因子
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We examined the conditions, kinetics and cell sources for basophil/mast cell and eosinophil colony stimulating activity (CSA) production by nasal epithelial cells taken from house dust (HD) nasal allergic patients. Epithelial scrapings removed from HD nasal allergic patients were grown to confluence over 7 days as a monolayer of epithelial cells in medium supplemented with fetal calf serum (FCS) on collagen coated microwell plates. CSA in nasal epithelial cells conditioned medium (CM) was assessed with density-gradient separated, nonadherent peripheral blood mononuclear cells in 14-day and 21-day methylcellulose assays. In the 14-day methylcellulose assays, the number of Eo-type colonies in the presence of either 10% or 5% CM was significantly higher than the background number of Eo-type colonies (negative control). Comparison of Gm CSA among 1%, 5% and 10% CM with negative, revealed no significant differences. We also compared the Eo-type CSA in the presence of 10% CM in the 14 -day methylcellulose assay with 21-day methylcellulose assay. There was no significant difference in the number of Eo-type colony between the 14-day and 21-day methylcellulose assays. We also examined the composition of the cells in the colonies in the 14-day and 21-day methylcellulose cultures. The percentage of metachromatic granule containing cells in a Eo-type colony in 14-day methylcellulose assay was significantly higher than in a Eo-type colony in 21-day methylcellulose assay. On the other hand, the percentage of eosinophilic granule containing cells in a Eo-type colony in 14-day methylcellulose assay was significantly lower than in a Eo-type colony in 21-day methylcellulose assay. We also measured histamine content in individual Eo-type colonies in 14-day and 21-day methylcellulose culture. Average histamine content in a Eo-type colony was 1.8±2. 2ng (N=15) in 14-day methylcellulose culture and 0. 7±0.8ng (N=15) in 21-day methylcellulose culture. These studies confirmed the existence of regulatory growth factors in the nasal epithelial cell CM taken from HD nasal allergic patients that stimulate and/or inhibit the growth and differentiation of human basophils/mast cells and eosinophils from common, committed progenitor cells and point to the existence of a unique microenvironment for basophil/mast cell and eosinophil development provided by nasal allergic epithelium.
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