Fasciola hepatica is refractory to complement killing by preventing attachment of mannose binding lectin (MBL) and inhibiting MBL-associated serine proteases (MASPs) with serpins

The complement system is a first-line innate host immune defence against invading pathogens. It is activated via three pathways, termed Classical, Lectin and Alternative, which are mediated by antibodies, carbohydrate arrays or microbial liposaccharides, respectively. The three complement pathways converge in the formation of C3-convertase followed by the assembly of a lethal pore-like structure, the membrane attack complex (MAC), on the pathogen surface. We found that the infectious stage of the helminth parasite Fasciola hepatica, the newly excysted juvenile (NEJ), is resistant to the damaging effects of complement. Despite being coated with mannosylated proteins, the main initiator of the Lectin pathway, the mannose binding lectin (MBL), does not bind to the surface of live NEJ. In addition, we found that recombinantly expressed serine protease inhibitors secreted by NEJ (rFhSrp1 and rFhSrp2) selectively prevent activation of the complement via the Lectin pathway. Our experiments demonstrate that rFhSrp1 and rFhSrp2 inhibit native and recombinant MBL-associated serine proteases (MASPs), impairing the primary step that mediates C3b and C4b deposition on the NEJ surface. Indeed, immunofluorescence studies show that MBL, C3b, C4b or MAC are not deposited on the surface of NEJ incubated in normal human serum. Taken together, our findings uncover new means by which a helminth parasite prevents the activation of the Lectin complement pathway to become refractory to killing via this host response, in spite of presenting an assortment of glycans on their surface. Author summaryThe mammalian complement system plays a central role in the defence against invasive pathogens. This response is initiated by recognition of specific molecules attached on the surface of bacteria, virus and parasites. Antigen-antibody complex, sugars and lipopolysaccharides are recognized by initiators of the Classical, Lectin and Alternative complement pathway, respectively, which leads to activation of the complement response and subsequent inflammation, opsonisation, damage and elimination of the invasive organism. Therefore, during infection with helminth parasites such as Fasciola hepatica, which is covered in antigenic sugars, it would be expected that complement attack, especially via the Lectin pathway, would eliminate them, thereby preventing establishment of infection. We discovered that the infective stage of F. hepatica, the newly excysted juvenile (NEJ), is resistant to the damaging effects of complement by using unique means that consists of: (1) preventing binding of the main initiator of this cascade, the mannose binding lectin (MBL), to the parasite surface; and (2) secreting two protease inhibitors (serpins), FhSrp1 and FhSrp2, that inactivate the main serine proteases involved in the Lectin pathway activation, namely MASPs. The elucidation of how F. hepatica and other helminths evade complement attack is a tractable approach for the discovery of novel anti-parasite interventions.