Antibody-dependent cellular cytotoxicity mediated by polymorphonuclear leukocytes and mononuclear cells against HSV-1 infected primary cultures of rabbit corneal epithelium.

Kwon BS, Colborn GL, Gangarosa LP, Hill JM

Abstract

The roles of rabbit polymorphonuclear leukocytes (PMNL) and mononuclear cells (MC) for the regulation of ocular herpes simplex virus-1 (HSV-1) infection were studied. The antibody-dependent cellular cytotoxicity (ADCC) mediated by PMNL and MC from normal rabbit peripheral blood was assessed kinetically employing a specific 51Cr release assay. The HSV-1 infected primary cultures of rabbit corneal epithelium (PRCE) were used as the target cells to obtain a homologous assay system. The PRCE was prepared by an epithelium outgrowth technique and identified by electron microscopy. The expression of the surface HSV-1 antigens on PRCE was examined by indirect immunofluorescent staining; the cell population stained by fluorescein increased from 40% at 3 hr postinfection (PI) to 100% at 8 hr PI. To determine how early the cytotoxicity occurs, PRCE were infected with HSV-1 for 2 hrs. After 2 hrs, the ADCC was checked every 10 min for the first 40 min and then at 1, 2 and 4 hr of incubation. The cytotoxicity was apparent at 10 min postincubation and reached 46% by PMNL and 40% by MC at 4 hr postincubation (6 hr PI). Significant cytotoxic effect (26% by PMNL and 16% by MC) occurred as early as 3 hr PI. When the one-step growth cycle of HSV-1 was studied in the PRCE, HSV-1 had an eclipse period of 4 hr and a rise period of 8 hr. This suggests that rabbit PMNL and MC have the potential to eliminate the HSV-1 infected rabbit corneal epithelium before HSV matures in the cells.

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