Inflammation Immunobiology Section, Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Eur J Immunol. 2013 Aug;43(8):2217-28. doi: 10.1002/eji.201343371. Epub 2013 Jun 14.
Here, we describe a novel method via which ex vivo cultured mouse bone marrow derived eosinophils (bmEos) can be adoptively transferred into recipient mice in order to study receptor-dependent recruitment to lung tissue in vivo. Intratracheal instillation of recombinant human eotaxin-2 (hCCL24) prior to introduction of bmEos via tail vein injection resulted in an approximately fourfold increase in Siglec F-positive/CD11c-negative eosinophils in the lungs of eosinophil-deficient ΔdblGATA recipient mice compared with controls. As anticipated, bmEos generated from CCR3-gene-deleted mice did not migrate to the lung in response to hCCL24 in this model, indicating specific receptor dependence. BmEos generated from GFP-positive BALB/c mice responded similarly to hCCL24 in vitro and were detected in lung tissue of BALB/c WT as well as BALB/c ΔdblGATA eosinophil-deficient recipient mice, at approximately fourfold (at 5 h post-injection) and approximately threefold (at 24 h postinjection) over baseline, respectively. Comparable results were obtained with GFP-positive C57BL/6 bmEos responding to intratracheal hCCL24 in C57BL/6 ΔdblGATA recipient mice. The use of ex vivo cultured bmEos via one or more of these methods offers the possibility of manipulating bmEos prior to transfer into a WT or gene-deleted recipient host. Thus, this chemotaxis model represents a novel and robust tool for pharmacological studies in vivo.
在这里,我们描述了一种新方法,通过该方法可以将体外培养的小鼠骨髓来源的嗜酸性粒细胞(bmEos)过继转移到受体小鼠中,以研究体内受体依赖性向肺组织募集。在通过尾静脉注射引入 bmEos 之前,向嗜酸性粒细胞缺陷型 ΔdblGATA 受体小鼠的气管内滴注重组人 eotaxin-2(hCCL24),导致 Siglec F 阳性/CD11c 阴性嗜酸性粒细胞在肺中的数量增加了约四倍与对照组相比。正如预期的那样,在该模型中,来自 CCR3 基因缺失小鼠的 bmEos 不会响应 hCCL24 迁移到肺部,表明存在特异性受体依赖性。从 GFP 阳性 BALB/c 小鼠生成的 bmEos 在体外对 hCCL24 有类似的反应,并在 BALB/c WT 以及 BALB/c ΔdblGATA 嗜酸性粒细胞缺陷型受体小鼠的肺组织中检测到,分别约为基线的四倍(注射后 5 小时)和约三倍(注射后 24 小时)。来自 GFP 阳性 C57BL/6 的 bmEos 对 C57BL/6 ΔdblGATA 受体小鼠气管内 hCCL24 的反应也得到了类似的结果。通过这些方法中的一种或多种使用体外培养的 bmEos,可以在将其转移到 WT 或基因缺失受体宿主之前对其进行操作。因此,该趋化性模型代表了体内药理学研究的一种新颖而强大的工具。