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对在粒细胞-巨噬细胞集落刺激因子或巨噬细胞集落刺激因子中培养的骨髓细胞来源的巨噬细胞中Ia抗原表达的分析。

Analysis of Ia antigen expression in macrophages derived from bone marrow cells cultured in granulocyte-macrophage colony-stimulating factor or macrophage colony-stimulating factor.

作者信息

Falk L A, Wahl L M, Vogel S N

机构信息

Department of Microbiology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814.

出版信息

J Immunol. 1988 Apr 15;140(8):2652-60.

PMID:3128601
Abstract

Bone marrow cells from C3H/HeJ mice were cultured in recombinant granulocyte-macrophage-CSF (rGM-CSF) or recombinant or purified (natural) preparations of macrophage-CSF (CSF-1) for 7 days, the adherent macrophages removed enzymatically, recultured in the absence of growth factor, and examined for their differentiative and functional characteristics. Expression of Ia Ag differed markedly in these two populations. Antibody plus complement-mediated cytotoxicity indicated that the percent of Ia-positive macrophages was low (approximately 15 to 20%) in both populations. Treatment of cultures with rIFN-gamma increased the percentage of Ia-positive cells in both populations; however, more CSF-1-derived macrophages were induced to become Ia positive than rGM-CSF-derived macrophages. In contrast, total Ia expression and Ia density per cell, as measured by ELISA and quantitative immunofluorescence, respectively, showed that medium-treated rGM-CSF-derived macrophages exhibited greater total Ia expression and higher Ia density per cell than CSF-1-derived macrophages. Treatment of CSF-1-derived macrophages with rIFN-gamma increased total Ia expression per culture to the levels exhibited by rGM-CSF-derived cells, although the density of Ia Ag per cell remained lower than in rGM-CSF-derived cells (medium or rIFN-gamma-treated). Northern blot and slot blot analysis of cytoplasmic RNA extracted from freshly harvested rGM-CSF- or CSF-1-derived cells indicated that the differences seen in basal Ia expression were also reflected at the level of steady-state, Ia-specific mRNA. The functional capacity of these two macrophage populations to stimulate Ag-specific T cell proliferation was also assessed. rGM-CSF- or CSF-1-derived macrophages were first cultured in the absence or presence of rIFN-gamma, Ag-pulsed, and irradiated before co-culture with Ag-primed, purified T cells. Ag-induced T cell proliferation was significantly greater in rGM-CSF- than in CSF-1-derived macrophages. Treatment of either population with rIFN-gamma had only a minimal effect on the ability of either macrophage population to stimulate Ag-specific T cell proliferation. These findings suggest that the development of mature macrophages from bone marrow progenitors under the influence of either GM-CSF or CSF-1 may, in part, underlie the functional heterogeneity of different macrophage populations.

摘要

将C3H/HeJ小鼠的骨髓细胞在重组粒细胞-巨噬细胞集落刺激因子(rGM-CSF)或巨噬细胞集落刺激因子(CSF-1)的重组或纯化(天然)制剂中培养7天,通过酶法去除贴壁巨噬细胞,在无生长因子的情况下重新培养,并检测其分化和功能特性。这两个群体中Ia抗原的表达有显著差异。抗体加补体介导的细胞毒性表明,两个群体中Ia阳性巨噬细胞的百分比都很低(约15%至20%)。用rIFN-γ处理培养物可增加两个群体中Ia阳性细胞的百分比;然而,与rGM-CSF来源的巨噬细胞相比,更多CSF-1来源的巨噬细胞被诱导成为Ia阳性。相反,分别通过ELISA和定量免疫荧光测量的总Ia表达和每个细胞的Ia密度显示,经培养基处理的rGM-CSF来源的巨噬细胞比CSF-1来源的巨噬细胞表现出更高的总Ia表达和更高的每个细胞的Ia密度。用rIFN-γ处理CSF-1来源的巨噬细胞可使每个培养物的总Ia表达增加到rGM-CSF来源细胞所表现出的水平,尽管每个细胞的Ia抗原密度仍低于rGM-CSF来源的细胞(培养基或rIFN-γ处理的)。对新鲜收获的rGM-CSF或CSF-1来源的细胞提取的细胞质RNA进行Northern印迹和狭缝印迹分析表明,在基础Ia表达中观察到的差异也反映在稳态Ia特异性mRNA水平上。还评估了这两个巨噬细胞群体刺激抗原特异性T细胞增殖的功能能力。rGM-CSF或CSF-1来源的巨噬细胞首先在无或有rIFN-γ的情况下培养,用抗原脉冲处理并照射,然后与抗原致敏的纯化T细胞共培养。抗原诱导的T细胞增殖在rGM-CSF来源的巨噬细胞中比在CSF-1来源的巨噬细胞中显著更大。用rIFN-γ处理任何一个群体对任何一个巨噬细胞群体刺激抗原特异性T细胞增殖的能力只有最小的影响。这些发现表明,在GM-CSF或CSF-1的影响下,骨髓祖细胞发育为成熟巨噬细胞可能部分是不同巨噬细胞群体功能异质性的基础。

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