title: Illuminating dynamic neutrophil trans-epithelial migration with micro-optical coherence tomography.
abstract: A model of neutrophil migration across epithelia is desirable to interrogate the underlying mechanisms of neutrophilic breach of mucosal barriers. A co-culture system consisting of a polarized mucosal epithelium and human neutrophils can provide a versatile model of trans-epithelial migration in vitro, but observations are typically limited to quantification of migrated neutrophils by myeloperoxidase correlation, a destructive assay that precludes direct longitudinal study. Our laboratory has recently developed a new isotropic 1-μm resolution optical imaging technique termed micro-optical coherence tomography (μOCT) that enables 4D (x,y,z,t) visualization of neutrophils in the co-culture environment. By applying μOCT to the trans-epithelial migration model, we can robustly monitor the spatial distribution as well as the quantity of neutrophils chemotactically crossing the epithelial boundary over time. Here, we demonstrate the imaging and quantitative migration results of our system as applied to neutrophils migrating across intestinal epithelia in response to a chemoattractant. We also demonstrate that perturbation of a key molecular event known to be critical for effective neutrophil trans-epithelial migration (CD18 engagement) substantially impacts this process both qualitatively and quantitatively.
authors.affiliation: Department of Dermatology, Harvard Medical School, Boston, MA, USA.|Wellman Center for Photomedicine, Boston, MA, USA.
authors.forename: Kengyeh K
authors.initials: KK
authors.lastname: Chu
authors.affiliation: Department of Pediatrics, Harvard Medical School, Boston, MA, USA.|Mucosal Immunology and Biology Research Center, Boston, MA, USA.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Kusek
authors.affiliation: Department of Dermatology, Harvard Medical School, Boston, MA, USA.|Wellman Center for Photomedicine, Boston, MA, USA.
authors.forename: Linbo
authors.initials: L
authors.lastname: Liu
authors.affiliation: Wellman Center for Photomedicine, Boston, MA, USA.|Mucosal Immunology and Biology Research Center, Boston, MA, USA.
authors.forename: Avira
authors.initials: A
authors.lastname: Som
authors.affiliation: Department of Pediatrics, Harvard Medical School, Boston, MA, USA.|Mucosal Immunology and Biology Research Center, Boston, MA, USA.
authors.forename: Lael M
authors.initials: LM
authors.lastname: Yonker
authors.affiliation: Department of Dermatology, Harvard Medical School, Boston, MA, USA.|Wellman Center for Photomedicine, Boston, MA, USA.
authors.forename: Huimin
authors.initials: H
authors.lastname: Leung
authors.affiliation: Department of Dermatology, Harvard Medical School, Boston, MA, USA.|Wellman Center for Photomedicine, Boston, MA, USA.
authors.forename: Dongyao
authors.initials: D
authors.lastname: Cui
authors.affiliation: Wellman Center for Photomedicine, Boston, MA, USA.
authors.forename: Jinhyeob
authors.initials: J
authors.lastname: Ryu
authors.affiliation: Mucosal Immunology and Biology Research Center, Boston, MA, USA.
authors.forename: Alexander D
authors.initials: AD
authors.lastname: Eaton
authors.affiliation: Department of Dermatology, Harvard Medical School, Boston, MA, USA.|Wellman Center for Photomedicine, Boston, MA, USA.|Department of Pathology, Massachusetts General Hospital, Boston, MA, USA.
authors.forename: Guillermo J
authors.initials: GJ
authors.lastname: Tearney
authors.affiliation: Department of Pediatrics, Harvard Medical School, Boston, MA, USA.|Mucosal Immunology and Biology Research Center, Boston, MA, USA.
authors.forename: Bryan P
authors.initials: BP
authors.lastname: Hurley
mesh_terms: D002448:Cell Adhesion; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte / Q000502:physiology; D018920:Coculture Techniques; D015179:Colorectal Neoplasms / Q000473:pathology*; D004848:Epithelium / Q000502:physiology*; D006801:Humans; D009504:Neutrophils / Q000166:cytology / Q000502:physiology*; D009195:Peroxidase / Q000378:metabolism*; D041623:Tomography, Optical Coherence / Q000379:methods*; D057705:Transendothelial and Transepithelial Migration
chemical_list: D009195:Peroxidase