Inhibitory effect of a complementary peptide on ulceration in the alkali-injured rabbit cornea.
OBJECTIVE
Two tripeptide chemoattractants, acetyl-proline-glycine-proline (Ac-PGP) and methyl-proline-glycine-proline (Me-PGP), are the primary triggers for early neutrophil invasion into the alkali-injured cornea. In the present study the effectiveness of a complementary peptide designed to inhibit the PGP chemoattractants (arginine-threonine-arginine [RTR] tetrameric peptide) and an apo A-1 mimicking peptide (5F) was investigated in the alkali-injured rabbit eye.


METHODS
(L)-RTR tetramer, (D)-RTR tetramer, and 5F were tested in vitro for their effects on neutrophil polarization. Synthetic 5F was also tested in vitro for its effect on the neutrophil respiratory burst. In the alkali-injured rabbit eye model, the right corneas of 48 rabbits were exposed to 1 N NaOH for 35 seconds. Sixteen animals were randomly assigned to each of three groups: phosphate-buffered saline (PBS) control; 800 microM RTR (dextrorotatory) tetramer in PBS alternating each hour with 1.5 mM RTR (levorotatory) tetramer in PBS; and 12 microM 5F in PBS. One topical drop of each substance was administered hourly (14 times per day) for 33 days. The experiment was continued until day 42 with no additional drops administered.


RESULTS
(L)-RTR tetramer and (D)-RTR tetramer inhibited neutrophil polarization activated by the PGP chemoattractants in vitro. Synthetic 5F did not inhibit neutrophil polarization in the presence of Ac-PGP or the respiratory burst of neutrophils in the presence of a metabolic stimulant derived from alkali-degraded corneas. During the entire animal experiment, statistically fewer ulcers occurred in the RTR tetramer group than in the PBS control group (43.8% vs. 87.5%, P = 0.0046). The frequency of ulceration in the 5F group (68.8%) was not significantly different from the PBS control group.


CONCLUSIONS
The reduction in the frequency of corneal ulceration by the RTR tetramer possibly resulted from its complementary binding to Ac-PGP and Me-PGP in the cornea shortly after alkali injury, leading to a reduction in the early and late infiltration of neutrophils. RTR tetramer appears to hold enough promise to warrant additional study as a therapeutic drug for the alkali-injured eye.
Haddox
J L
JL
Eye Research Laboratories, Sight Savers of Alabama, Birmingham 35209, USA. sightsavers@mindspring.com
Pfister
R R
RR
Sommers
C I
CI
Blalock
J E
JE
Villain
M
M
D000818:Animals; D016375:Antisense Elements (Genetics) / Q000627:therapeutic use; D016632:Apolipoprotein A-I / Q000737:chemistry; D002057:Burns, Chemical / Q000276:immunology / Q000517:prevention & control*; D002630:Chemotactic Factors / Q000037:antagonists & inhibitors*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D000529:Complementary Therapies; D003315:Cornea / Q000187:drug effects / Q000276:immunology; D003320:Corneal Ulcer / Q000139:chemically induced / Q000276:immunology / Q000517:prevention & control*; D005126:Eye Burns / Q000139:chemically induced*; D016231:Fluorescence Polarization Immunoassay; D009504:Neutrophils / Q000502:physiology*; D009842:Oligopeptides / Q000037:antagonists & inhibitors / Q000138:chemical synthesis / Q000627:therapeutic use*; D011392:Proline / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors; D011817:Rabbits; D016897:Respiratory Burst / Q000187:drug effects; D012972:Sodium Hydroxide
D016375:Antisense Elements (Genetics); D016632:Apolipoprotein A-I; D002630:Chemotactic Factors; D009842:Oligopeptides; C113164:prolyl-glycyl-proline; D012972:Sodium Hydroxide; D011392:Proline