title: Chemotaxis of bone marrow derived eosinophils in vivo: a novel method to explore receptor-dependent trafficking in the mouse.
abstract: 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.
authors.affiliation: Inflammation Immunobiology Section, Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA. eva.sturm@medunigraz.at
authors.forename: Eva M
authors.initials: EM
authors.lastname: Sturm
authors.forename: Kimberly D
authors.initials: KD
authors.lastname: Dyer
authors.forename: Caroline M
authors.initials: CM
authors.lastname: Percopo
authors.forename: Akos
authors.initials: A
authors.lastname: Heinemann
authors.forename: Helene F
authors.initials: HF
authors.lastname: Rosenberg
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology; D039521:CD11c Antigen / Q000096:biosynthesis; D002478:Cells, Cultured; D054423:Chemokine CCL24 / Q000494:pharmacology; D002634:Chemotaxis, Leukocyte / Q000276:immunology*; D004804:Eosinophils / Q000166:cytology / Q000276:immunology* / Q000637:transplantation; D049452:Green Fluorescent Proteins / Q000235:genetics; D008168:Lung / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D054397:Receptors, CCR3 / Q000235:genetics
chemical_list: C515484:CCL24 protein, human; D039521:CD11c Antigen; C515351:Ccr3 protein, mouse; D054423:Chemokine CCL24; D054397:Receptors, CCR3; D049452:Green Fluorescent Proteins
keywords: Bone morrow derived eosinophils; CCL24; Chemotaxis; Immunology; Mouse model