1. Angiogenesis in Viral Immunoinflammatory Lesions Barry T. Rouse University of Tennessee Part 1

2. The Smokies’ Lost Tribe “Of course we do not have herpes”

3. Herpetic Stromal Keratitis An immunoinflammatory reaction in the eye that appears to represent a Type 1 CD4+ T cell mediated reaction in a tissue, the cornea, that normally lacks a vascular system and has been considered as one site of immunological privilege.

5. HSV Keratitis in humans Most common infectious cause of blindness in USA At least 400,000 cases About 20% of cases have the immunoinflammatory stromal form Requires chronic treatment and may lead to corneal transplantation

6. Pathogenesis of Stromal Keratitis Complex involves multiple humoral and cellular participants Neovascularization a major feature Principal orchestrators of the lesion are CD4+ T cells of the type 1 phenotype

7. HSK is caused by CD4+ type 1 T cells but IL-12 expression in the eye prior to HSV infection diminishes rather than exaggerates the lesions. -Lee et. Al. 2002

8. 0 25 50 75 100 05101520 IL-12 DNA vector DNA 0 1 2 3 4 5 05101520 Expression of IL-12 DNA prior to infection inhibits HSK severity and incidence Incidence (%) Severity Days post infection

9. How does the IL-12 effect function? Antiviral effect via IFN-  - No Immunosuppression via iNOS - No Modulation of immunity - No Indirect effect on angiogenesis

10.  -actin IP-10 IFN-  MIG IL-10 Gene expression in cornea at day 3 post infection by RT PCR BALB/c GKO naive Virus + vector Virus + IL-12 Virus + vector Virus + IL-12

11. 0 5 10 15 20 Angiogenic scoring 05101520 Days post infection IL-12 DNA IP-10 DNA vector DNA IL-12 and IP-10 DNA both inhibit ocular angiogenesis IL-12 DNA effect reversed by anti IP-10 + anti MIG

12. INTERPRETATION IL-12 expression causes upregulation via IFN-  induction of IP- 10 (and MIG). These chemokines bind to heparan sulfate and block the angiogenic effect of FGF. Consequence : inhibited angiogenesis which could be necessary to develop stromal keratitis following virus infection.

13. Which molecules are induced in the cornea that can account for HSV-driven neovascularization? Is preventing angiogenesis a logical therapeutic goal for herpetic keratitis?

14. Viruses- neovascularization VirusLesionMechanism HHV8Kaposi’s SarcomaEncodes multiple angiogenic proteins HBVHepatitisActivates HIF-1a that turns on VEGF promoter HIVKaposi’s SarcomaTat activates VEGF-R2 HPVCervical CancerIncreases VEGF via indirect complex mechanism HSVHerpetic Stromal Keratitis Mysterious but unraveling

15. Productive Infection By HSV Most host mRNA and protein synthesis rapidly shut down A few host proteins are made and secreted. These include IL-1 and IL-6.

16. To continue open the Part 2 from Supercourse-Vet htpp://centaur.vri.cz