Hieronymus Thomas, Gust Tatjana C, Kirsch Ralf D, Jorgas Thorsten, Blendinger Gitta, Goncharenko Mykola, Supplitt Kamilla, Rose-John Stefan, Müller Albrecht M, Zenke Martin
Institute for Biomedical Engineering-Cell Biology, University Medical School Rheinisch-Westfälische Technische Hochschule Aachen, Aachen, Germany.
J Immunol. 2005 Mar 1;174(5):2552-62. doi: 10.4049/jimmunol.174.5.2552.
Dendritic cells (DC) represent key regulators of the immune system, yet their development from hemopoietic precursors is poorly defined. In this study, we describe an in vitro system for amplification of a Flt3(+)CD11b(+) progenitor from mouse bone marrow with specific cytokines. Such progenitor cells develop into both CD11b(+) and CD11b(-) DC, and CD8alpha(+) and CD8alpha(-) DC in vivo. Furthermore, with GM-CSF, these progenitors synchronously differentiated into fully functional DC in vitro. This two-step culture system yields homogeneous populations of Flt3(+)CD11b(+) progenitor cells in high numbers and allows monitoring the consecutive steps of DC development in vitro under well-defined conditions. We used phenotypic and functional markers and transcriptional profiling by DNA microarrays to study the Flt3(+)CD11b(+) progenitor and differentiated DC. We report here on an extensive analysis of the surface Ag expression of Flt3(+)CD11b(+) progenitor cells and relate that to surface Ag expression of hemopoietic stem cells. Flt3(+)CD11b(+) progenitors studied exhibit a broad overlap of surface Ags with stem cells and express several stem cell Ags such as Flt3, IL-6R, c-kit/SCF receptor, and CD93/AA4.1, CD133/AC133, and CD49f/integrin alpha(6). Thus, Flt3(+)CD11b(+) progenitors express several stem cell surface Ags and develop into both CD11b(+) and CD11b(-) DC, and CD8alpha(+) and CD8alpha(-) DC in vivo, and thus into both of the main conventional DC subtypes.
树突状细胞（DC）是免疫系统的关键调节因子，但其从造血前体的发育过程尚不清楚。在本研究中，我们描述了一种体外系统，可利用特定细胞因子从小鼠骨髓中扩增Flt3(+)CD11b(+)祖细胞。这种祖细胞在体内可发育为CD11b(+)和CD11b(-) DC，以及CD8α(+)和CD8α(-) DC。此外，在粒细胞-巨噬细胞集落刺激因子（GM-CSF）作用下，这些祖细胞在体外可同步分化为功能完全的DC。这种两步培养系统可大量产生均一的Flt3(+)CD11b(+)祖细胞群体，并允许在明确的条件下体外监测DC发育的连续步骤。我们使用表型和功能标记以及DNA微阵列进行转录谱分析，以研究Flt3(+)CD11b(+)祖细胞和分化的DC。我们在此报告对Flt3(+)CD11b(+)祖细胞表面抗原表达的广泛分析，并将其与造血干细胞的表面抗原表达相关联。所研究的Flt3(+)CD11b(+)祖细胞与干细胞在表面抗原上有广泛重叠，并表达几种干细胞抗原，如Flt3、白细胞介素-6受体、c-kit/干细胞因子受体，以及CD93/AA4.1、CD133/AC133和CD49f/整合素α(6)。因此，Flt3(+)CD11b(+)祖细胞表达几种干细胞表面抗原，在体内可发育为CD11b(+)和CD11b(-) DC，以及CD8α(+)和CD8α(-) DC，从而发育为两种主要的传统DC亚型。
