Distribution and Quantification of Choroidal Macrophages in Human Eyes With Age-Related Macular Degeneration.
OBJECTIVE
Increasing evidence suggests a role for macrophages in the pathogenesis of age-related macular degeneration (AMD). This study examined choroidal macrophages and their activation in postmortem eyes from subjects with and without AMD.


METHODS
Choroids were incubated with anti-ionized calcium-binding adapter molecule 1 (anti-IBA1) to label macrophages, anti-human leukocyte antigen-antigen D-related (anti-HLA-DR) as a macrophage activation marker, and Ulex europaeus agglutinin lectin to label blood vessels. Whole mounts were imaged using confocal microscopy. IBA1- and HLA-DR-positive (activated) cells were counted in submacula, paramacula, and nonmacula, and cell volume and sphericity were determined using computer-assisted image analysis.


RESULTS
In aged control eyes, the mean number of submacular IBA1+ and HLA-DR+ macrophages was 433/mm2 and 152/mm2, respectively. In early AMD eyes, there was a significant increase in IBA1+ and HLA-DR+ cells in submacula compared to those in controls (P = 0.0015 and P = 0.008, respectively). In eyes with neovascular AMD, there were significantly more HLA-DR+ cells associated with submacular choroidal neovascularization (P = 0.001). Mean cell volume was significantly lower (P ≤ 0.02), and sphericity was significantly higher (P ≤ 0.005) in all AMD groups compared to controls.


CONCLUSIONS
The average number of IBA1+ macrophages in submacular and paramacular choroid was significantly higher in early/intermediate AMD compared to that in aged controls. HLA-DR+ submacular macrophages were significantly increased in all stages of AMD, and they were significantly more round and smaller in size in the submacular AMD choroid, suggesting their activation. These findings support the concept that AMD is an inflammatory disease.
Wilmer Ophthalmological Institute, Johns Hopkins Hospital, Baltimore, Maryland.
D Scott
DS
McLeod
Wilmer Ophthalmological Institute, Johns Hopkins Hospital, Baltimore, Maryland.
Imran
I
Bhutto
Wilmer Ophthalmological Institute, Johns Hopkins Hospital, Baltimore, Maryland.
Malia M
MM
Edwards
Ophthalmic Epidemiology and Genetics Service, New England Eye Center, Tufts Medical Center, Boston, Massachusetts.
Rachel E
RE
Silver
Ophthalmic Epidemiology and Genetics Service, New England Eye Center, Tufts Medical Center, Boston, Massachusetts 3Department of Ophthalmology, Tufts University School of Medicine, Boston, Massachusetts.
Johanna M
JM
Seddon
Wilmer Ophthalmological Institute, Johns Hopkins Hospital, Baltimore, Maryland.
Gerard A
GA
Lutty
D000368:Aged; D000369:Aged, 80 and over; D016022:Case-Control Studies; D002452:Cell Count; D002829:Choroid / Q000473:pathology*; D020256:Choroidal Neovascularization / Q000473:pathology*; D005260:Female; D006801:Humans; D008264:Macrophages / Q000473:pathology*; D008268:Macular Degeneration / Q000473:pathology*; D008297:Male