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E. Normando, B. Stampachiacchiere, R. Sgrulletta, A. Micera, A. Lambiase, S. Bonini; Evidence for a Modulatory Action of NGF on Toll–Like Receptor Expression by Primary Culture of Ocular Cicatricial Pemphigoid–Derived Fibroblasts . Invest. Ophthalmol. Vis. Sci. 2006;47(13):4942.
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Toll like receptors (TLR) play a key role in innate immunity, promoting efficient immune responses through the interaction between the innate and adaptive immune system and between immune and resident cells (including fibroblasts). Nerve growth factor (NGF), other than affecting a variety of immune cells, modulates conjunctival fibroblasts activity. The purpose of this study is to investigate, at molecular and biochemical levels, changes of TLR–4 and TLR–9 expression by ocular cicatricial pemphigoid (OCP)–derived fibroblasts (FBs) compared to healthy ones, and to evaluate whether NGF influences the TLRs expression by OCP–derived FBs.
primary culture of OCP–derived FBs (n=3) and healthy FBs (n=3) were evaluated for TLR–4 and 9 mRNA and proteins expression by relative real time–PCR and confocal analysis respectively. Moreover, healthy and OCP–derived FBs were exposed to different concentrations of NGF (from 1 to 100 ng/ml) and changes of TLR–4 and TLR–9 mRNA and protein expression were evaluated. Statistical analysis was performed using ANOVA test.
: OCP–derived FBs showed a marked increase of TLR–4 mRNA expression (86 fold increase) when compared to healthy derived FBs (p<0.05). A lower increase of TLR–9 mRNA (8 fold) was observed in OCP–derived FBs (p<0.05); molecular data are confirmed by confocal analysis. Short term (24hrs) exposure to increasing concentration of NGF significantly modulate TLR–4 and 9 expression by both healthy–derived and OCP–derived FBs. NGF exposure significantly decreased TLR–4 mRNA expression in a dose dependent fashion; by the way only low or high concentration of NGF significantly decreased TLR–9 mRNA expression whereas 10ng/ml NGF induced a significant increase of TLR–9.
In OCP–derived FBs, compared to healthy FBs, both TLR4 and 9 mRNA/protein expression appear up–regulated. Moreover, NGF seems to influence the expression of TLR 4 in a dose–dependent way.
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