Flow cytometric identification of a minority population of MHC class II positive cells in the normal rat retina distinct from CD45lowCD11b/c+CD4low parenchymal microglia
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AIMS: This study aimed to isolate and classify by flow cytometry, the cell surface phenotype of microglia in the normal rat retina with a view to identifying putative antigen presenting cells (APC) within the retina, which has to date not been possible by immunohistochemistry. METHODS: Normal rat retinal microglia were isolated and classified using a modification of an isolation technique employing graduated Percoll density gradient cell separation and flow cytometric phenotypic criteria used for CNS microglia. RESULTS: Retinal microglia can be defined by flow cytometry on the basis of their CD45lowCD11b/c+CD4low cell surface expression. Constitutive MHC class II expression in the normal rat retina was confined almost exclusively to a very minor population of cells expressing neither low (microglia) nor high levels of CD45. Three colour flow cytometric analysis confirmed that these MHC class II positive cells were ED2+. CONCLUSIONS: Using this sensitive isolation technique we have identified the cell surface characteristics of ramified, resident microglia, and found that they do not constitutively express MHC class II. There is, however, constitutive MHC class II expression on a phenotypically distinct population of cells (CD45low/highED2+). We propose these cells are the counterpart of the perivascular macrophages found in the CNS which present antigen to extravasating T cells, although their exact retinal location can only be confirmed by immunohistochemical analysis. The role of parenchymal microglia as APC remains undefined. Future isolation of microglia and putative perivascular cells using this technique will help identify the role these cells play in the initiation and perpetuation of immune responses within the retina.
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Co-expression of CD45 and CD11b has effectively been used to distinguish microglia and macrophages.
“However, although the co-expression of CD45 and CD11b has effectively been used to distinguish microglia and macrophages,17, 18, 19, 20,49 microglia can increase CD45 expression following activation, which may limit the resolution between these two populations in the inflamed retina.50,51”
From Characterizing the cellular immune response to subretinal AAV gene therapy in the murine retina (2021) · cites this paper as [49]
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Self-citation
- Authors
- Dick, A. D., Ford, A. L., Forrester, J. V., & Sedgwick, J. D.
- Affiliations
- Centenary Institute (AU)
- Venue
- British Journal of Ophthalmology
- Publisher
- BMJ
- Published
- 1995-09-01
- Volume
- 79
- Issue
- 9
- Pages
- 834-840
- ISSN
- 0007-1161 (ISSN-L)
- PMC
- PMC505270
- OpenAlex
- W2078901795
- Cited by
- 129
- References
- 26
- Citations by year
- 2026: 4 · 2025: 2 · 2024: 3 · 2023: 1 · 2021: 15 · 2020: 4 · 2019: 2 · 2018: 7 · 2017: 3 · 2016: 4 · 2015: 7 · 2014: 5 · 2013: 2 · 2012: 4
- FWCI
- 1.64
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- British Journal of Ophthalmology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- UCL Discovery (University College London) (submittedVersion)
- HighWire Press Open Archive (submittedVersion)
- Funders
- Wellcome Trust
- Topics
- Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
- Health Sciences › Medicine › Ophthalmology › Ocular Diseases and Behçet’s Syndrome
- Health Sciences › Medicine › Ophthalmology › Glaucoma and retinal disorders
- Keywords
- retinal microglia, MHC class II expression, antigen-presenting cells, ED2, CD45, CD11b, perivascular macrophages, retinal immune response, microglia isolation, rat retina
- MeSH terms
- Animals, Central Nervous System, Flow Cytometry, Major Histocompatibility Complex, Phenotype, Rats, Inbred Lew, Retina, CD4 Antigens, Integrin alphaXbeta2, Macrophage-1 Antigen, Leukocyte Common Antigens, Microglia, Rats
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