Abstract
Recently, we reported that a native Fasciola hepatica fatty acid binding protein (FABP) termed Fh12 is a powerful anti-inflammatory protein capable of suppressing the LPS-induced expression of inflammatory markers in vivo and in vitro. Because the purification of a protein in native form is, in many situations not cost-beneficial and unsuitable for industrial grade scale-up, this study accomplished the task of optimizing the expression and purification of a recombinant form of FABP (Fh15). Additionally, we ascertained whether this molecule could exhibit a similar suppressive effect on TLR-stimulation and inflammatory cytokine expression from macrophages than those previously demonstrated for the native molecule. Results demonstrated that Fh15 suppresses the expression of IL-1β and TNFα in murine macrophages and THP1 Blue CD14 cells. Additionally, Fh15 suppress the LPS-induced TLR4 stimulation. This effect was not impaired by a thermal denaturing process or blocked by the presence of anti-Fh12 antibodies. Fh15 also suppressed the stimulation of various TLRs in response to whole bacteria extracts, suggesting that Fh15 could have a broad spectrum of action. These results support the possibility of using Fh15 as an excellent alternative for an anti-inflammatory drug in preclinical studies in the near future.
Highlights
Fatty acid binding proteins (FABPs) in platyhelminths constitute a multigenic family of cytoplasmic proteins with isoforms localized in tegument and parenchymal cells
We recently reported that a native 12 kDa member of the F. hepatica FABP (Fh12) significantly suppresses the cytokine storm and other inflammatory mediators induced by lipopolysaccharide (LPS), which is the potent endotoxin of Gram-negative bacteria
Because E. coli is the first choice of host when a protein has to be expressed, the main goal of this study was to optimize the expression of a recombinant FABP termed Fh15 in E. coli, and determine whether Fh15 is able to mimic the anti-inflammatory properties showed by Fh12
Summary
Fatty acid binding proteins (FABPs) in platyhelminths constitute a multigenic family of cytoplasmic proteins with isoforms localized in tegument and parenchymal cells. Parasitic trematodes are unable to synthesize lipids de novo, in particular long-chain fatty acids and cholesterol[1, 2] They use carriers to uptake such lipids directly from the host and transport them to specific destinations within parasite, a process in which FABP could play an important role. During the infection, F. hepatica antigens, having FABP as a constituent could be saturating CD14 located on the surface of macrophages making them refractory to subsequent stimuli. Based on this particular mode of action, we considered that Fh12 is an attractive molecule with potential to develop a drug against sepsis, UC or any other inflammatory disease in which TLR4 is involved. These results support the possibility of testing Fh15 as an anti-inflammatory drug in preclinical studies in the near future
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