In vivo quantification of leukocyte behavior in the retina during endotoxin-induced uveitis.
OBJECTIVE
The interaction between leukocytes and vascular endothelial cells plays an important role in various inflammatory disorders. This study evaluated leukocyte behavior in the retina during endotoxin-induced uveitis (EIU) in vivo.


METHODS
EIU was induced in female Lewis rats by footpad injection of lipopolysaccharide (LPS). The time-course changes of retinal leukocyte behavior were followed at 1.5, 3, 4.5, 6, 12, 24, 48, 72, and 120 hours after LPS treatment using acridine orange digital fluorography, consisting of high-resolution images from a scanning laser ophthalmoscope and a fluorescent nuclear dye of acridine orange.


RESULTS
Major retinal vessels were significantly dilated (P < 0.05) at 4.5 hours after LPS injection. The vasodilation, marked in veins, became maximum at 24 hours and subsided at 72 hours. Leukocytes were observed rolling along the walls of major veins at 4.5 hours. The number of rolling leukocytes gradually increased and reached a peak level of 33.8 +/- 3.4 cells/minute per major vein at 12 hours. Leukocyte rolling was still observed at 72 hours. No rolling of leukocytes was observed along the arterial walls throughout any experiments. The velocities of rolling leukocytes were determined at 6, 12, 24, and 48 hours. The leukocyte rolling velocity at 6 hours was significantly slower (33.3 +/- 2.8 microns/second, P < 0.05) than at the other three times (average, 46.6 microns/second). Cellular infiltration into the vitreous cavity was detected at 24 hours and reached its maximum at 48 hours.


CONCLUSIONS
This study demonstrates that it is possible to evaluate EIU by investigating retinal leukocyte behavior and that vasodilation of major retinal vessels and leukocyte-endothelial interactions precede inflammatory cell emigration into the vitreous. This method may be useful to quantify the severity of inflammation in EIU.
Miyamoto
K
K
Department of Ophthalmology and Visual Sciences, Kyoto University Graduate School of Medicine, Japan.
Ogura
Y
Y
Hamada
M
M
Nishiwaki
H
H
Hiroshiba
N
N
Honda
Y
Y
D000165:Acridine Orange; D000818:Animals; D001082:Aqueous Humor / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D004731:Endotoxins / Q000633:toxicity*; D005136:Eye Proteins / Q000378:metabolism; D005260:Female; D005451:Fluorescein Angiography / Q000379:methods; D005456:Fluorescent Dyes; D005471:Fluoroscopy; D007091:Image Processing, Computer-Assisted; D007834:Lasers; D007958:Leukocyte Count; D007962:Leukocytes / Q000166:cytology / Q000502:physiology*; D019721:Ophthalmoscopes; D051381:Rats; D011917:Rats, Inbred Lew; D012171:Retinal Vessels / Q000473:pathology / Q000503:physiopathology*; D012486:Salmonella typhimurium; D015866:Uveitis, Posterior / Q000139:chemically induced / Q000473:pathology / Q000503:physiopathology*
D004731:Endotoxins; D005136:Eye Proteins; D005456:Fluorescent Dyes; D000165:Acridine Orange