Improvement of HSV-1 necrotizing keratitis with amniotic membrane transplantation.
OBJECTIVE
Stromal herpes simplex virus keratitis (HSK) is an immune-mediated disease. Previous studies have indicated that T cells, neutrophils, and macrophages contribute to the tissue damage in HSK. It has been shown that human amniotic membrane promotes epithelial wound healing and has diverse anti-inflammatory effects. In this study, the effect of amniotic membrane transplantation (AMT) on corneal wound healing and on inflammation in mice with necrotizing HSK was examined.


METHODS
BALB/c mice were corneally infected with 10(5) plaque-forming units (PFU) of HSV-1 (KOS strain). In 16 mice that exhibited severe ulcerating HSK, the cornea was covered with a preserved human amniotic membrane as a patch. Corneas in 16 infected mice remained uncovered and served as a control. On days 2 and 7 after surgery, the amniotic membrane was removed (eight mice in each group), the HSV-1-infected cornea was evaluated clinically, and the eye was enucleated. Tissue sections were analyzed histologically for epithelialization and cellular infiltration and immunohistochemically with anti-CD3 mAb to T cells, anti-CD11b mAb to both macrophages and neutrophils, or anti-F4/80 mAb to macrophages.


RESULTS
Profound regression of corneal inflammation and rapid closure of epithelial defects were observed clinically within 2 days in the amniotic membrane-covered eyes, whereas HSV-1 keratitis and ulceration progressed in all mice in the control group (P < 0.001). Histologically, corneal edema and inflammatory infiltration, and immunohistochemically the number of CD3(+), CD11b(+), and F4/80(+) cells in the cornea were markedly decreased at 2 and 7 days after amniotic membrane application, compared with the uncovered control corneas (P < 0.001).


CONCLUSIONS
AMT promotes rapid epithelialization and reduces stromal inflammation and ulceration in HSV-1 keratitis. AMT in mice with HSV necrotizing stromal keratitis appears to be a useful model for investigating the effect and the action mechanism of human amniotic membrane.
Heiligenhaus
A
A
Department of Ophthalmology, University of Essen, Germany. arnd.heligenhaus@t-online.de
Bauer
D
D
Meller
D
D
Steuhl
K P
KP
Tseng
S C
SC
D000650:Amnion / Q000637:transplantation*; D000818:Animals; D017252:CD3 Complex / Q000032:analysis; D003315:Cornea / Q000276:immunology / Q000382:microbiology / Q000601:surgery*; D018259:Herpesvirus 1, Human / Q000502:physiology*; D006801:Humans; D007124:Immunoenzyme Techniques; D016849:Keratitis, Herpetic / Q000276:immunology / Q000382:microbiology / Q000601:surgery*; D016177:Macrophage-1 Antigen / Q000032:analysis; D008264:Macrophages / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009504:Neutrophils / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D014945:Wound Healing
D017252:CD3 Complex; D016177:Macrophage-1 Antigen