|Budget Amount *help
¥2,400,000 (Direct Cost : ¥2,400,000)
Fiscal Year 1999 : ¥1,200,000 (Direct Cost : ¥1,200,000)
Fiscal Year 1998 : ¥1,200,000 (Direct Cost : ¥1,200,000)
1. Human neutrophil chemotaxis induced by herpes simplex virus (HSV) infection
Using in-vitro system with human corneal epithelial cells immortalized with SV-40, human corneal stromal cells, human neutrophils, and Boyden chamber, human neutrophil chemotaxis was found to be induced by HSV-infected corneal epithelial cells and stromal cells after infection of HSV via interleukin-8.
2. Cytokine production from human peripheral blood T lymphocytes of herpetic keratitis patients
(1) T lymphocytes and antigen-presenting cells primed with UV-inactivated HSV and irradiated with γ-ray, culture media were collected, and the amounts of cytokines were measured with ELISA.
(2) T lymphocytes of herpetic keratitis patients produced Th1 cytokines such as IFN-γ and IL-2, but not Th2 cytokines (actually minimal level of IL-10 and under-detection level of IL-4)
(3) Patients with herpetic recurrences of stromal type within 1 year produced higher level of Th1 cytokines and indicated significantly higher ratio o
f IFN-γ/IL-10 and IL-2/IL-10 than those with herpetic recurrences of epithelial type within 1 year.
3. The efficacy of herpes vaccine in murine herpetic keratitis model
To investigate the utility of immune therapy with herpes vaccine, we constructed chimera-protein consisted of HSV glycoprotein-D and human IL-2, and study its efficacy using primary corneal HSV infection model in mice.
(1) Systemic and local (subconjunctival) administration of gD-IL-2 protein inhibited the development of stromal type of herpetic keratitis.
(2) Subconjunctival injection of gD-IL-2 DNA induced humoral and cellular immunity against HSV.
(3) Subconjunctival injection of gD-IL-2 DNA inhibited the development of stromal type of herpetic keratitis.
(4) Subconjunctival injection of gD-IL-2 DNA suppressed viral proliferation in eyes, trigeminal ganglia, and brains.
4. Immunohistochemical study in murine herpetic keratitis model
Using murine herpetic keratitis model, cytokines, adhesion molecules, infiltrated cells, and major histocompatibility antigens in cornea and local lymph nodes were immunohistochemically studied, and dense localization of Th1 cytokines and γδ-T cells was ascertained. Less