antigen-presenting cells
DESCRIPTION
Use of Intravital Multi-Photon Microscopy to Study In Vivo Migratory Kinetics of Corneal Bone Marrow-Derived Cells Pedram Hamrah, M.D. , Dimosthenis Mantopoulos, MD; Lixin Zheng, MD; Ulrich von Andrian, MD, PhD - PowerPoint PPT PresentationTRANSCRIPT
Use of Intravital Multi-Photon Microscopy to Study In Vivo Migratory Kinetics of Corneal
Bone Marrow-Derived Cells
Pedram Hamrah, M.D.Pedram Hamrah, M.D., Dimosthenis Mantopoulos, MD; Lixin Zheng, MD; Ulrich von Andrian, MD, PhD
Massachusetts Eye & Ear Infirmary, Department of Ophthalmology & Immune Massachusetts Eye & Ear Infirmary, Department of Ophthalmology & Immune Disease Institute, Harvard Medical SchoolDisease Institute, Harvard Medical School
Financial Disclosure: The authors have no financial disclosure related to this projectFinancial Disclosure: The authors have no financial disclosure related to this project
Support: Support: NEI K12-EY016335, New England Corneal Transplant Research Fund, Falk Medical Research Trust
Antigen-presenting Cells
Sentinels of the immune system
Dendritic cells, macrophages and B cells
Dendritic Cells and macrophages are the
professional antigen-presenting cells
(APC) of the cornea
Implicated in corneal transplantation and
allergic immunity, microbial keratitis, and
dry eye disease
Corneal, unlike limbal, APC are universally MHC class II-negative and immature in the epithelium and stroma
Limbus
Green = Class II (Maturation marker)
Limbus
Red = CD45 (Bone marrow marker)
Green = Class II
Periphery
Red = CD11c(DC marker)
Green = CD80 (B7 costimulatory marker)
Center
Red = CD11c
Green = CD80
Hamrah et al., Novel Characterization of MHC class II-negative Population of Resident Corneal Langerhans cell-type Dendritic Cells. Invest Ophthalmol Vis Sci, 2002; 43:639-646
Hamrah et al., The Corneal Stroma is Endowed with Significant Numbers of Resident Dendritic Cells. Invest Ophthalmol Vis Sci, 2003; 44:581-589
Hamrah and Dana, Immune homeostasis of the eye: Antigen Presenting Cells in the Eye and Ocular Surface. Encyclopedia of the Eye. Elsevier. In press
Advantages of Multi-Photon Microscopy
Deeper imaging into scattering specimens
Reduced out of plane photobleaching and
photodamage in optically thick specimens
Access to nonlinear signals other than
fluorescence such as second harmonic
scattering
Purpose and Methods
The purpose of this study was to dissect migratory properties of
antigen presenting cells by novel multi-photon intravital microscopy.
Multi-photon intravital microscopy (MPM) of the cornea was applied
to investigate localization and trafficking properties of corneal APCs
in transgenic mice in steady state and inflammation in vivo.
CD11c-eYFP
MHC class II-eGFPDay 5 Inflammation
MHC class II-eGFPDay 5 inflammation
Normal Inflamed
Results Intravital MPM studies of the normal cornea demonstrated that
APCs were sparsely distributed centrally and more dense in the
periphery
Epithelial and stromal APCs were distinguished by second
harmonic generation that visualizes stromal collagen
While APCs demonstrated continuous sampling motions in steady
state, cells generally did not migrate laterally
During inflammation, increased numbers of APCs were
demonstrated, exhibiting extreme morphological changes
An increase in lateral and vertical migration was shown particularly
in stromal subpopulations
Conclusions Our studies are the first to demonstrate long-term migratory kinetics of corneal
APCs in steady state and inflammation through high-resolution intravital multi-
photon microscopy.
Collectively, these models allow for dissecting molecular regulation of APC
recruitment to, and migration in the cornea.