ALZET 2001D osmotic pumps, one containing mouse anti–ICAM-1 IgG2a
mAb (2 mg/ml) from clone BIRR0001 and one containing mouse nonimmune
IgG2a mAb (2 mg/ml) (R&D Systems, Minneapolis, MN), were implanted in
the superotemporal quadrant of each eye. The surgeon was masked to the
identity of the two pumps. Six hours after implantation, animals were
anesthetized, and 0.5% proparacaine (Alcon, Ft. Worth, TX) and 0.3%
ofloxacin (Allergan, Hormigueros, PR) eye drops were topically applied.
After pump placement, 2 μg of human recombinant VEGF
(VEGF165), diluted in 100 μl of pyrogen-free Dulbecco’s phosphate-buffered saline (PBS; Sigma),
was injected into the vitreous body through the inferonasal pars plana
of each eye with a 30-gauge needle. To normalize intraocular pressure,
100 μl of aqueous humor was removed with a 30-gauge needle. Animals
were killed 24 hours after implantation and MPO activity was measured
in ocular tissues. To ensure that the intravitreous injection did not
provide an intraocular conduit for the antibodies, 2 animals were
implanted with ALZET 2001D osmotic pumps containing FITC–IgG, and
a 30-gauge needle was used to perforate the inferonasal sclera. The
fluorescence in ocular tissues, 24 hours later, was compared with that
in animals without the perforation.