Differential expression of secretory granule proteases in mouse mast cells exposed to interleukin 3 and c-kit ligand.
Differential expression of secretory granule proteases in mouse mast cells exposed to interleukin 3 and c-kit ligand.
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DOI：
10.1084/jem.175.4.1003
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发表时间：
1992-04-01
期刊：
影响因子：
--
通讯作者：
Stevens RL
中科院分区：
文献类型：
--
作者：
Gurish MF;Ghildyal N;McNeil HP;Austen KF;Gillis S;Stevens RL
现在已经确定，存在于粘膜和浆膜环境中的肥大细胞（MC）亚类可以根据其特异性分泌颗粒定位蛋白酶和蛋白多糖的表达来相互区分。此外，现在可以使用最近开发的基因特异性探针和重组细胞因子来鉴定调节编码颗粒这些成分的基因表达的造血和结缔组织来源的细胞因子。当用重组白细胞介素3 （il -3）培养骨髓源性MC (BMMC)，并在缺乏或存在重组c-kit配体（rKL）的情况下用该细胞因子维持时，它们仍然是红素-，几乎不产生35s标记的肝素蛋白聚糖，并且含有比MMCP-6 mRNA更高水平的小鼠MC蛋白酶(MMCP) -5 mRNA和肥大细胞羧肽酶A (MC- cpa) mRNA。它们不含MMCP-4或-2 mRNA，这两种基因分别在祖细胞分化为浆膜和粘膜MCs的后期表达。相比之下，BMMC与rKL单独或在含有rIL-3的培养基中依次培养，rKL表达高水平的MMCP-4和-6 mRNA，以及编码MMCP-5和MC-CPA的转录本。尽管rkl开发的BMMC是红花素+，并产生大量的35s标记的肝素蛋白多糖，但相对于浆膜MC，它们只含有极少量的组胺和MC- cpa酶活性。这些研究首次表征了使用任何重组细胞因子衍生的BMMC群体的转录颗粒表型，以证明组织化学染色和颗粒成熟之间的解离。并证明细胞因子对晚表达蛋白酶基因的拮抗调节作用。
It is now established that the subclasses of mast cells (MC) that reside in mucosal and serosal environments can be distinguished from one another in terms of their expression of specific secretory granule- localized proteases and proteoglycans. Further, the hematopoietic- and connective tissue-derived cytokines that regulate expression of the genes that encode these constituents of the granule can now be identified using recently developed gene-specific probes and recombinant cytokines. When bone marrow-derived MC (BMMC) were developed with recombinant interleukin 3 (rIL-3) and maintained with this cytokine in the absence or presence of recombinant c-kit ligand (rKL), they remained safranin-, produced almost no 35S-labeled heparin proteoglycans, and contained greater levels of mouse MC protease (MMCP) -5 mRNA and mast cell carboxypeptidase A (MC-CPA) mRNA than MMCP-6 mRNA. They did not contain MMCP-4 or -2 mRNA, genes expressed late in the differentiation of progenitor cells into serosal and mucosal MCs, respectively. In contrast, BMMC developed with rKL alone or by sequential culture in medium containing rIL-3 followed by rKL expressed high levels of MMCP-4 and -6 mRNA, as well as the transcripts that encode MMCP-5 and MC-CPA. Although rKL-developed BMMC were safranin+ and produced substantial amounts of 35S-labeled heparin proteoglycans, they contained only minimal amounts of histamine and MC-CPA enzymatic activity relative to serosal MC. These are the first studies to characterize the transcriptional granule phenotype of a population of BMMC derived using any recombinant cytokine, to demonstrate a dissociation between histochemical staining and granule maturation, and to demonstrate antagonistic regulation of late expressed protease genes by a cytokine.