Increased severity of HSV-1 keratitis and mortality in mice lacking the 2-5A-dependent RNase L gene.
OBJECTIVE
The2',5'-oligoadenylate-dependent RNase L gene functions in the interferon-inducible RNA decay pathway known as the 2-5A system. The purpose of this study was to determine whether the absence of this gene affects the pathogenesis of herpes simplex virus type 1 (HSV-1) ocular infection in the mouse.


METHODS
HSV-1 (strain McKrae) was applied bilaterally to unscarified corneas of RNase L-null mice and congenic controls. To evaluate the severity of herpetic keratitis, slit lamp examinations (SLE) were performed every other day for 14 days. To study corneal histology and apoptosis, HSV-1-inoculated RNase-L-null and congenic control mice, as well as mock-inoculated mice (apoptosis negative control), were killed at 6 and 18 hours postinoculation (PI). Uninoculated mice that underwent corneal scarification (apoptosis positive control) were killed 2 hours after scarification. Eyes were dissected and the corneas processed for light and transmission electron microscopy and the TUNEL assay.


RESULTS
In comparison with the congenic control mice, RNase L-null mice showed significantly more severe herpetic keratitis (PI day 8, SLE score, mean +/- SEM: 3.27 +/- 0.10 vs. 2.34 +/- 0.06; P: < 0.001) and significantly higher mortality (PI day 14, 70% vs. 20%; P: < 0.001). Few apoptotic cells were seen in HSV-1-infected RNase L-null mice, although DNA fragmentation consistent with apoptosis was detected in the corneas of congenic control mice 6 and 18 hours after HSV-1 inoculation and in uninfected mice with scarified corneas. Signs of apoptosis were not present in the mock-infected corneas. Electron microscopic evidence of keratocytic apoptosis was detected only in the uninfected scarified corneas and the HSV-1-infected congenic control corneas.


CONCLUSIONS
The increased severity of ocular disease and increased mortality in the RNase L-null mice provides evidence, for the first time, that the 2-5A system contributes to protection during ocular herpetic infection. The reduced frequency of apoptosis in these mice suggests that one possible mechanism for this protective effect could be the induction of apoptosis in corneal cells as a means of reducing the spread of infectious virus.
Zheng
X
X
Department of Ophthalmology, LSU Eye Center, Louisiana State University Health Sciences Center, New Orleans, Louisiana, USA.
Silverman
R H
RH
Zhou
A
A
Goto
T
T
Kwon
B S
BS
Kaufman
H E
HE
Hill
J M
JM
D000818:Animals; D003315:Cornea / Q000648:ultrastructure / Q000821:virology*; D053938:DNA Fragmentation; D004722:Endoribonucleases / Q000502:physiology*; D018259:Herpesvirus 1, Human / Q000472:pathogenicity*; D020287:In Situ Nick-End Labeling; D016849:Keratitis, Herpetic / Q000201:enzymology / Q000401:mortality* / Q000473:pathology / Q000821:virology*; D051379:Mice; D018345:Mice, Knockout; D014774:Virulence; D014779:Virus Replication
D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease