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Production1
Department of Anatomy/Cell Biology, Wayne State University School of Medicine, Detroit, MI 48201
Pseudomonas aeruginosa keratitis destroys the cornea
in susceptible (B6), but not resistant (BALB/c) mice. To determine
mechanisms mediating resistance, the role of IFN-
, IL-12, and IL-18
was tested in BALB/c mice. RT-PCR analysis detected IFN- mRNA
expression levels in cornea that were significantly increased at 1–7
days postinfection. IL-18 mRNA was detected constitutively in cornea
and, at 1–7 days postinfection, levels were elevated significantly,
while no IL-12 mRNA was similarly detected. To test whether IL-18
contributed to IFN- production, mice were treated with
anti-IL-18 mAb. Treatment decreased corneal IFN- mRNA levels,
and bacterial load and disease increased/worsened, compared with
IgG-treated mice. To stringently examine the role of IFN- in
bacterial killing, knockout (-/-) vs wild-type (wt) mice also were
tested. All corneas perforated, and bacterial load was increased
significantly in -/- vs wt mice. Because disease severity was
increased in IFN--/- vs IL-18-neutralized mice,
and since IL-18 also induces production of TNF, we tested for TNF-
in both groups. ELISA analysis demonstrated significantly elevated
corneal TNF- protein levels in IFN--/- vs wt mice
after infection. In contrast, RT-PCR analysis of IL-18-neutralized vs
IgG-treated infected mice revealed decreased corneal TNF- mRNA
expression. Next, to resolve whether TNF was required for bacterial
killing, TNF- was neutralized in BALB/c mice. No difference in
corneal bacterial load was detected in neutralized vs IgG-treated mice.
These data provide evidence that IL-18 contributes to the resistance
response by induction of IFN- and that IFN- is required for
bacterial killing.
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