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Subcutaneous injection of type 2 herpes simplex virus (HSV-2) (10(3) PFU) to newborn rabbits produced severe skin lesions and wide dissemination of the virus to various organs including the eye. Ocular lesions were characterized by retinal folds and choroiditis. HSV could be isolated from mononuclear cells (MNCs) of infected animal blood. Newborn rabbits treated with monoclonal antibody (MAb) against glycoprotein D (gD) of HSV on days 0, 2 and 4 postinfection had little or no skin lesions (0.0-2.3 mm) compared to controls (2.8-13.0 mm). In addition, the MAb treatment significantly suppressed dissemination of the virus to the eye (0% in MAb-treated vs 83% in control) and other organs and reduced the rate of chorioretinitis (0% in MAb-treated vs 50% in control). The treatment of HSV-infected MNCs with MAb resulted in 91-100% reduction in infectivity of the cells. The results suggest that anti-gD MAb protects newborn rabbits from HSV-2 eye infection by neutralizing the virus in skin and inactivating HSV-infected MNCs in blood.

To isolate and characterize fetal trabecular meshwork (FTM) cells for study in culture. Cultured adult trabecular meshwork (TM) cells often possess a slower rate of growth and restricted number of population doublings, limiting the ability to perform expanded testing.Fetal eyes from 24-week gestation abortions were delicately dissected to isolate the developing trabecular meshwork tissue. Three primary cultures were achieved and passaged. Light microscopy was used to compare the FTM cells to two cultured adult TM cell lines. Immunocytochemistry and Western blot analysis were utilized to identify specific protein expression.The FTM cells demonstrated similar microscopic characteristics to adult TM cells, including monolayer formation, cobblestone pattern, and comparable size. FTM cells exhibited faster, more consistent doubling times when compared with adult TM cells. They grew rapidly even after passage 8, whereas their adult counterparts slowed significantly with each successive passage and failed to reach confluence at passages 4 to 5. Immunofluorescent staining was positive for actin, vimentin, fibronectin, laminin, aquaporin-1, CD-44, and myocilin in both FTM and adult TM cells. In both fetal and adult cells, Western blots showed substantial increase in myocilin after exposure to dexamethasone.Characterization by microscopy and immunocytochemistry suggest that FTM cells have properties similar to adult TM cells. Fetal tissues may be a useful source of abundant, rapidly dividing FTM cells for in vitro investigation. The ability to do expanded research in this field may contribute to a better understanding of the molecular mechanisms in glaucoma development.

We studied the protective effects of passive immunization with virus specific antibody in newborn rabbits inoculated subcutaneously with type 2 herpes simplex virus (HSV-2). Newborn rabbits given anti-HSV-2 antibody intraperitoneally (IP) on days 0, 2 and 4 post infection had smaller herpetic skin lesions and reduced mortality when compared to controls. In addition, the IP treatment using this schedule reduced virus growth in the skin lesions and virus dissemination, so that it decreased the frequency of herpetic retinitis. When the IP antibody administration was started at 24 hours post virus inoculation, according to the schedule days 1, 3 and 5, there was less protection; larger skin lesions, higher mortality, and greater evidence of virus dissemination. Also HSV-infected mononuclear cells (MNCs) treated with anti-HSV serum resulted in a significant reduction in the number of infected MNCs. The results of these studies suggest that anti-HSV-2 antibody contributes to protection against HSV-2 infection of skin as well as eyes, probably by inactivation of the virus locally at the skin inoculation site, and by combating the hematogenous spread of HSV-infected MNCs as well as free virus to various organs including the eye.