Basu Sunanda, Zhang Hui-Hua, Quilici Cathy, Dunn Ashley R
Ludwig Institute for Cancer Research, Melbourne Tumor Biology Branch, Royal Melbourne Hospital, Victoria, Australia.
Stem Cells Dev. 2004 Feb;13(1):39-50. doi: 10.1089/154732804773099245.
Previously, we have reported that although unperturbed granulocyte colony-stimulating factor (GCSF)-deficient (G-CSF-/-) mice are neutropenic, when challenged with Candida albicans, they develop a profound neutrophilia. In an attempt to understand the basis of Candida-induced neutrophilia in G-CSF-deficient mice, we have modified the Dexter bone marrow culture system to produce an in vitro model that mimics emergency granulopoiesis in vivo. In this model, stromal cultures are overlaid with bone marrow cells in the presence or absence of heat-inactivated (HI) Candida. Irrespective of the genotype of mice used as a source of bone marrow-derived stromal cells, stimulation of these cultures with HI Candida led to a significantly greater recovery of cells compared to unstimulated stromal cultures. In addition, there was a marked increase in the number of colony-forming units granulocyte-macrophage (CFU-GM), as well as in the percentage of granulocytes in the population of nonadherent cells recovered from HI Candida-stimulated cultures. The conditioned medium generated from stromal cultures derived from either wild-type or G-CSF-/- mice exposed to HI Candida, when applied to bone marrow cells in a soft agar clonogenic assay stimulated M-, GM-, and G- type colonies. Interleukin-3 (IL-3) and GM-CSF could not be detected in the conditioned medium from either HI Candida stimulated or unstimulated stromal cultures. However, IL-6 was detected in the conditioned media from both wild-type and G-CSF-/- stromal cultures. Addition of anti-IL-6 antibody significantly impaired granulopoiesis in unstimulated and HI Candida-stimulated, wild type, and G-CSF-/- stromal cultures. Conditioned medium generated from G-CSF/IL-6-deficient stromal cells had the capacity to stimulate bone marrow cells to form colonies comprised of granulocytes and macrophages in soft agar clonogenic assay. This study demonstrates that stromal cells can be stimulated with HI Candida and gives an insight into Candida mediated granulopoiesis.
此前，我们曾报道，尽管未受干扰的粒细胞集落刺激因子（GCSF）缺陷（G-CSF-/-）小鼠存在中性粒细胞减少症，但在用白色念珠菌攻击时，它们会出现严重的中性粒细胞增多。为了理解G-CSF缺陷小鼠中念珠菌诱导的中性粒细胞增多的基础，我们对德克斯特骨髓培养系统进行了改良，以建立一个体外模型，该模型可模拟体内的应急粒细胞生成。在这个模型中，在存在或不存在热灭活（HI）念珠菌的情况下，将骨髓细胞覆盖在基质培养物上。无论用作骨髓来源的基质细胞的小鼠基因型如何，与未刺激的基质培养物相比，用HI念珠菌刺激这些培养物会导致细胞回收率显著更高。此外，集落形成单位粒细胞-巨噬细胞（CFU-GM）的数量以及从HI念珠菌刺激的培养物中回收的非贴壁细胞群体中粒细胞的百分比均显著增加。从暴露于HI念珠菌的野生型或G-CSF-/-小鼠的基质培养物中产生的条件培养基，在软琼脂克隆形成试验中应用于骨髓细胞时，可刺激M-、GM-和G-型集落。在HI念珠菌刺激或未刺激的基质培养物的条件培养基中均未检测到白细胞介素-3（IL-3）和粒细胞-巨噬细胞集落刺激因子（GM-CSF）。然而，在野生型和G-CSF-/-基质培养物的条件培养基中均检测到了IL-6。添加抗IL-6抗体显著损害了未刺激和HI念珠菌刺激的野生型和G-CSF-/-基质培养物中的粒细胞生成。从G-CSF/IL-6缺陷的基质细胞产生的条件培养基有能力在软琼脂克隆形成试验中刺激骨髓细胞形成由粒细胞和巨噬细胞组成的集落。这项研究表明，HI念珠菌可刺激基质细胞，并深入了解念珠菌介导的粒细胞生成。
