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从体内肥大细胞缺陷的小鼠体外获得的小鼠骨髓来源的肥大细胞(mBMMC)表达与来自其正常同窝小鼠的mBMMC和浆膜肥大细胞相同的一组颗粒蛋白酶。

Mouse bone marrow-derived mast cells (mBMMC) obtained in vitro from mice that are mast cell-deficient in vivo express the same panel of granule proteases as mBMMC and serosal mast cells from their normal littermates.

作者信息

Eklund K K, Ghildyal N, Austen K F, Friend D S, Schiller V, Stevens R L

机构信息

Department of Medicine, Harvard Medical School, Boston, Massachusetts.

出版信息

J Exp Med. 1994 Jul 1;180(1):67-73. doi: 10.1084/jem.180.1.67.

Abstract

The ear, skin, and purified serosal mast cells of WBB6F1/J-(+/+) (WB-(+/+)) and WCB6F1/J-(+/+) (WC-(+/+)) mice contain high steady-state levels of the transcripts that encode mouse mast cell protease (mMCP) 2, mMCP-4, mMCP-5, mMCP-6, and mouse mast cell carboxypeptidase A (mMC-CPA). In contrast, no mast cell protease transcripts are present in abundance in the ear and skin of WBB6F1/J-W/Wv (W/Wv) and WCB6F1/J-Sl/Sld (Sl/Sld) mice which are mast cell-deficient in vivo due to defects in their c-kit and c-kit ligand genes, respectively. We now report that the immature bone marrow-derived mast cells (mBMMC) obtained in vitro with recombinant interleukin 3 (rIL-3) or WEHI-3 cell conditioned medium from WB-(+/+), WC-(+/+), W/Wv, and Sl/Sld mice all contain high steady-state levels of the mMCP-2, mMCP-4, mMCP-5, mMCP-6, and mMC-CPA transcripts. As assessed immunohistochemically, mMCP-2 protein and mMCP-5 protein are also present in the granules of mBMMC from WB-(+/+), WC-(+/+), and W/Wv mice. That Sl/Sld and W/Wv mBMMC contain high steady-state levels of five granule protease transcripts expressed by the mature serosal, ear, and skin mast cells of their normal +/+ littermates suggests that c-kit-mediated signal transduction is not essential for inducing transcription of these protease genes. Because rIL-4 inhibits the rIL-10-induced expression of mMCP-1 and mMCP-2 in BALB/cJ mBMMC, the ability of rIL-4 to influence protease mRNA levels in WC-(+/+) mBMMC and W/Wv mBMMC was investigated. Although rIL-10 induced expression of the mMCP-1 transcript in WC-(+/+) and W/Wv mBMMC, rIL-4 was not able to suppress the steady-state levels of the mMCP-1 transcript or any other protease transcript in these cultured mast cells. Thus, not only do BALB/cJ mBMMC express fewer granule proteases than mBMMC from mast cell-deficient strains and their normal littermates but the subsequent induction of late-expressed proteases in BALB/cJ mBMMC is more tightly regulated by IL-3 and IL-4.

摘要

WBB6F1/J-(+/+)(WB-(+/+))和WCB6F1/J-(+/+)(WC-(+/+))小鼠的耳朵、皮肤及纯化的浆膜肥大细胞中,编码小鼠肥大细胞蛋白酶(mMCP)2、mMCP-4、mMCP-5、mMCP-6和小鼠肥大细胞羧肽酶A(mMC-CPA)的转录本处于较高的稳态水平。相比之下,WBB6F1/J-W/Wv(W/Wv)和WCB6F1/J-Sl/Sld(Sl/Sld)小鼠的耳朵和皮肤中,肥大细胞蛋白酶转录本并不丰富,这两种小鼠由于其c-kit和c-kit配体基因存在缺陷,在体内缺乏肥大细胞。我们现在报告,用重组白细胞介素3(rIL-3)或来自WB-(+/+)、WC-(+/+)、W/Wv和Sl/Sld小鼠的WEHI-3细胞条件培养基在体外获得的未成熟骨髓来源肥大细胞(mBMMC),均含有高水平的mMCP-2、mMCP-4、mMCP-5、mMCP-6和mMC-CPA转录本。免疫组织化学评估显示,WB-(+/+)、WC-(+/+)和W/Wv小鼠的mBMMC颗粒中也存在mMCP-2蛋白和mMCP-5蛋白。Sl/Sld和W/Wv的mBMMC含有其正常+/+同窝小鼠成熟浆膜、耳朵和皮肤肥大细胞表达的五种颗粒蛋白酶转录本的高水平稳态,这表明c-kit介导的信号转导对于诱导这些蛋白酶基因的转录并非必不可少。由于rIL-4抑制BALB/cJ mBMMC中rIL-10诱导的mMCP-1和mMCP-2表达,因此研究了rIL-4影响WC-(+/+) mBMMC和W/Wv mBMMC中蛋白酶mRNA水平的能力。尽管rIL-10诱导了WC-(+/+)和W/Wv mBMMC中mMCP-1转录本的表达,但rIL-4无法抑制这些培养的肥大细胞中mMCP-1转录本或任何其他蛋白酶转录本的稳态水平。因此,不仅BALB/cJ mBMMC表达的颗粒蛋白酶比来自肥大细胞缺陷品系及其正常同窝小鼠的mBMMC少,而且BALB/cJ mBMMC中晚期表达蛋白酶的后续诱导受IL-3和IL-4的调控更为严格。

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