CNL, a ricin B-like lectin from mushroom Clitocybe nebularis, induces maturation and activation of dendritic cells via toll-like receptor 4 pathway
Švajger, Urban ...
A novel lectin, isolated from basidiomycete mushroom Clitocybe nebularis and termed C. nebularis lectin (CNL), exhibits an immunostimulatory effect on the most potent antigen presenting cells, the ...dendritic cells (DCs). Treatment of human monocyte-derived DCs with CNL in doses from 1-10 [micro]g/ml resulted in a dose-dependent induction of overall DC maturation characteristics. Exposure of DCs to CNL for 48h resulted in extensive up-regulation of co-stimulatory molecules CD80, CD86, as well as the maturation marker CD83 and HLA-DR molecules. Such CNL-matured DCs (CNL-DCs) were capable of inducing a Th1-polarized response in naive CD4+CD45RA+ T cells in 5-day allogeneic co-cultures. The allostimulatory potential of CNL-DCs was significantly increased relative to untreated controls, as was their capacity to produce several pro-inflammatory cytokines such as IL-6, IL-8 and TNF-[alpha]. By using a specific TLR4 signaling inhibitor, CLI-095, as well as Myd88 inhibitory peptide, we have shown that DC activation by CNL is completely dependent on the TLR4 activation pathway. Furthermore, activation of TLR4 by CNL was confirmed via TLR4 reporter assay. Measurement of p65 Nf-kB and p38 MAPK phosphorylation levels following CNL stimulation of DCs revealed primarily an increase in Nf-kB activity, with less effect on the induction of p38 MAPK signaling than that of LPS-matured DCs. CNL had the ability to activate human DCs in such a way as to subsequently direct Th1-type T cell responses. Our results encourage the use of mushroom-derived lectins for use in therapeutic strategies, with aims such as to strengthen anti-tumor immune responses.
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