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  • 1
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Mast cells (MCs) arise from haematopoietic stem cells. We have recently reported that CD34+ progenitors derived from human bone marrow (BM) develop into tryptase+, chymase+ MCs when cultured in the presence of recombinant human stem cell factor (rhSCF) and recombinant human IL-6 (rhIL-6). In an experiment for the expression of chymase during differentiation, chymase+ cells were detected in human BM, but tryptase+ cells were not found.Objective The purpose of this study was to show the appearance of chymase+ cells in CD34+ cells with an origin different from MC differentiation.Methods CD34+ cells from human BM were sorted with anti-CD117 monoclonal antibody (mAb), and cytospins of CD34+, CD34+CD117+, or CD34+CD117− were prepared. These cells were cultured with rhSCF+rhIL-6 for 12 weeks. Some of the cells were subjected to either histological stain with Wright–Giemsa or immunocytochemistry with anti-chymase mAb. Real-time RT-PCR was also performed to compare the transcriptional level of chymase from each cell preparation.Results Chymase was expressed in CD34+ cells as well as human MCs by immunocytochemistry. Substantial CD34+CD117− cells, but not CD34+CD117+ cells, were stained immunocytochemically with anti-chymase mAb. For 1 week culture with rhSCF+rhIL-6, no cells expressed chymase in any preparation. Real-time RT-PCR revealed positivity for chymase mRNA in CD34+ cells, but it reduced at 1 week of culture, and increased as cells developed into MCs. Chymase mRNA in CD34+CD117+ cells was negligible compared with that in CD34+CD117−. Tryptase mRNA was below the detectable level in CD34+ cells, and increased along with MC differentiation. After 12 weeks of culture, CD34+CD117+ developed predominantly into MCs, whereas CD34+CD117− developed into monocytes/macrophages.Conclusion Our findings suggested that chymase is present not only in MCs but also in CD34+CD117− BM progenitors, but that its origin is different from the MC lineage.
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  • 2
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background CD34+ progenitor cells develop into tryptase+, CD117+ mast cells when cultured in the presence of recombinant human stem cell factor (rhSCF). However, spontaneous IgE receptor (FcɛRI) expression during human mast cell development is not well examined.Objective Here, the expression and function of FcɛRI in and on human bone marrow-derived mast cells (HBMMCs) during development were investigated.Methods and Results At 4 weeks of culture, predominant cells expressed high-affinity IgE receptor α chain (FcɛRIα) on the cell surface determined by flow cytometry, but CD117 was less expressed. Immunocytochemistry with antitryptase mAb and anti-FcɛRIα mAb revealed intracellular and surface expression of FcɛRIα at 2 weeks of culture, but tryptase was less expressed. FcɛRIα mRNA transcript preceded that of tryptase mRNA at 2 weeks of culture determined by real-time RT-PCR, and FcɛRIα, FcɛRIβ, FcɛRIγ, and tryptase mRNA increased along with differentiation. FcɛRIα cross-link on HBMMC and 4-week-old mast cells/mast cell precursors induced the release of IL-5 and granulocyte macrophage-colony stimulating factor, which was enhanced by rhSCF.Conclusion These data indicated that HBMMC constitutively and spontaneously expressed functional FcɛRI subunits at the early stage of differentiation, probably because of the differences in the ability and functional property of progenitors.
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  • 3
    ISSN: 1420-9071
    Keywords: Key words.klotho; endothelial function; nitric oxide production; hypertension; renal failure; cytokine.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The human aging process is associated with vascular endothelial dysfunction. However, humoral factors which might protect against endothelial dysfucntion during aging have not yet been identified. We recently identified the klotho gene as a possible regulator of human aging. In the present study using the klotho-deficient heterozygous mouse, we examined whether the Klotho protein is a humoral factor protecting against endothelial dysfunction. We further cloned rat klotho cDNA and investigated whether klotho mRNA expression in rat kidney is altered under pathological conditions such as hypertension, hyperlipidemia, renal failure, and inflammatory stress. The Klotho protein itself, or its metabolites, promotes endothelial NO production in aorta as well as arterioles, and klotho mRNA in kidney is downregulated under sustained circulatory stress.
    Type of Medium: Electronic Resource
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