Effect of monoclonal antibodies on monocyte trafficking in retinal vessels in EAU. Blocking antibodies to CD62L, CD44, and PSGL-1 significantly reduced rolling and sticking of monocytes in inflamed vessels. Freshly isolated EGFP bone marrow–derived monocytes (8 × 106) were injected intravenously into mice immunized 21 to 24 days previously with peptide to induce EAU. After 48 hours, cell trafficking in the retinal vasculature was analyzed by SLO. Retinal images were recorded for 15 minutes, and then mice were injected intravenously with 30 μg/mouse of rat anti–mouse antibody, and recording continued for a further 20 minutes. Data were then compared before and after antibody treatment. (A) Recirculation of adoptively transferred cells was expressed as the number of transferred EGFP monocytes detected in the same section of the retinal vessel before and after antibody infusion. (B) Rolling efficiency, expressed as the percentage of rolling fluorescent cells among the total number of fluorescent cells entering a venule before and after antibody infusion. (C) Sticking efficiency, expressed as the percentage of fluorescent monocytes within the same venule that remained adherent for at least 20 seconds. *P < .05; **P < .01; Student paired t test; n was at least 16 vessels from 3 mice.
