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首页> 外文期刊>Allergy >Alteration and acquisition of Siglecs during in vitro maturation of CD34+ progenitors into human mast cells.
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Alteration and acquisition of Siglecs during in vitro maturation of CD34+ progenitors into human mast cells.

机译：CD34 +祖细胞体外成熟进入人肥大细胞期间，Siglecs的改变和获得。

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Using human mast cells (MC) derived by culture of CD34+ peripheral blood precursors, a comprehensive study was performed of expression of 11 known Siglecs. Analysis was initially performed at the mRNA level using gene arrays. Positive results were then validated at the protein level using indirect immunofluorescence and flow cytometry, and for some Siglecs, Western blot analysis was also used. Culture-derived MC expressed mRNA for CD22 (Siglec-2), CD33 (Siglec-3), Siglec-5, Siglec-6, Siglec-8 and Siglec-10. Flow cytometry confirmed surface expression of all these molecules except for CD22 and Siglec-10, where levels were low or undetectable. However, Western blotting was able to detect MC expression of CD22 and Siglec-10, suggesting that these proteins were mostly cytoplasmic. CD34+ precursor cells from peripheral blood constitutively expressed surface CD33, Siglec-5 and Siglec-10. As they matured into MC, their constitutive levels of CD33 changed little, Siglec-5 and Siglec-10 declined, and Siglec-6 and Siglec-8 appeared de novo, all in parallel with accumulation of histamine and other MC markers, such as surface expression of FcepsilonRIalpha, and CD51. Phenotypic analysis of LAD-2 MC yielded a similar pattern of Siglec expression except that CD22 expression was particularly prominent. Finally, immunohistochemistry confirmed expression of these same Siglecs by mature tryptase-positive MC in human lung tissues. These data demonstrate an extensive and previously unappreciated pattern of Siglec expression on human MC. Whether engagement and signaling through these inhibitory Siglecs can impact MC biology will require further investigation.
机译：使用通过培养CD34 +外周血前体细胞衍生的人肥大细胞（MC），进行了11种已知Siglecs表达的全面研究。最初使用基因阵列在mRNA水平进行分析。然后，使用间接免疫荧光和流式细胞术在蛋白质水平上验证了阳性结果，对于某些Siglecs，还使用了蛋白质印迹分析。培养来源的MC表达CD22（Siglec-2），CD33（Siglec-3），Siglec-5，Siglec-6，Siglec-8和Siglec-10的mRNA。流式细胞仪证实除CD22和Siglec-10以外，所有这些分子的表面表达都很低，后者水平较低或无法检测到。但是，蛋白质印迹能够检测CD22和Siglec-10的MC表达，表明这些蛋白主要是细胞质的。来自外周血的CD34 +前体细胞组成性表达表面CD33，Siglec-5和Siglec-10。当它们成熟为MC时，其CD33的组成水平变化不大，Siglec-5和Siglec-10下降，并且从头出现Siglec-6和Siglec-8，与组胺和其他MC标记物（如表面）的积累平行FcepsilonRIalpha和CD51的表达。 LAD-2 MC的表型分析产生了类似的Siglec表达模式，只是CD22表达特别突出。最后，免疫组化证实了成熟的类胰蛋白酶阳性的MC在人肺组织中也表达了这些相同的Siglecs。这些数据证明了在人MC上Siglec表达的广泛且以前未被认识的模式。通过这些抑制性Siglecs的参与和信号传导是否会影响MC生物学，还需要进一步研究。

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