Quantitative Analysis of Transforming Growth Factor-beta1 in Human Fibroblasts Induced with Staphylococcus enterotoxin B and Lipopolysaccharide

Lee, Seong Geun; Kim, Kwang Hyuk; Kim, Uk Kyu; Kim, Jong Ryoul; Chung, In Kyo; Yang, Dong Kyu

Abstract

TGF-beta1 is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to infection control. The objective of this study is to investigate production of TGF-beta in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of TGF-beta1 which may be responsible for infection control. The fibroblasts were originated from gingiva and facial dermis in 26 year-old male patient. In the presence of LPS(0.0lmicrogram, 0.1microgram, 1.0microgram), SEB(0.0lmicrogram, 0.1microgram, 1.0microgram) respectively, cells(5x103ml) were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, cells(2.5x105ml) were cultivated in EMEM with LPS(0.01, 0.1 and 1.0microgram), SEB(0.01, 0.1 and 1.0microgram) respectively and LPS(0.1microgram) and SEB(0.1microgram) in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and TGF-beta1 was assayed in duplicate. The results were as follows. 1. In gingival fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly since 3 days after incubation, compared with the control and the production of TGF-beta1 occurred very significantly at 1 day after incubation, compared with the control. 2. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of TGF-beta1 was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of TGF-beta1 was increased very significantly at 1 day after incubation, compared with the control. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of TGF-beta1 very significantly. The gingival and facial dermal fibroblasts have different phenotype each other The orchestrated understanding of fibroblast proliferation and TGF-beta1 production play an important part in host defense against the bacterial infection and may prevent tissue necrosis such as necrotizing fasciitis and life-threatening syndrome such as multiple organ failure.