BackgroundSchistosomiasis, a severe disease caused by parasites of the genus Schistosoma, is prevalent in 74 countries, affecting more than 250 million people, particularly children. We have previously shown that the Schistosoma mansoni gut-derived cysteine peptidase, cathepsin B1 (SmCB1), administered without adjuvant, elicits protection (>60%) against challenge infection of S. mansoni or S. haematobium in outbred, CD-1 mice. Here we compare the immunogenicity and protective potential of another gut-derived cysteine peptidase, S. mansoni cathepsin L3 (SmCL3), alone, and in combination with SmCB1. We also examined whether protective responses could be boosted by including a third non-peptidase schistosome secreted molecule, glyceraldehyde 3-phosphate dehydrogenase (SG3PDH), with the two peptidases.Methodology/Principal findingsWhile adjuvant-free SmCB1 and SmCL3 induced type 2 polarized responses in CD-1 outbred mice those elicited by SmCL3 were far weaker than those induced by SmCB1. Nevertheless, both cysteine peptidases evoked highly significant (P < 0.005) reduction in challenge worm burden (54–65%) as well as worm egg counts and viability. A combination of SmCL3 and SmCB1 did not induce significantly stronger immune responses or higher protection than that achieved using each peptidase alone. However, when the two peptidases were combined with SG3PDH the levels of protection against challenge S. mansoni infection reached 70–76% and were accompanied by highly significant (P < 0.005) decreases in worm egg counts and viability. Similarly, high levels of protection were achieved in hamsters immunized with the cysteine peptidase/SG3PDH-based vaccine.Conclusions/SignificanceGut-derived cysteine peptidases are highly protective against schistosome challenge infection when administered subcutaneously without adjuvant to outbred CD-1 mice and hamsters, and can also act to enhance the efficacy of other schistosome antigens, such as SG3PDH. This cysteine peptidase-based vaccine should now be advanced to experiments in non-human primates and, if shown promise, progressed to Phase 1 safety trials in humans.

No abstract

Graphical abstract

Several reports have documented the reproducible and considerable efficacy of the cysteine peptidase-based schistosomiasis vaccine in the protection of mice and hamsters against infection with Schistosoma mansoni and Schistosomahaematobium, respectively. Here, we attempt to identify and define the protection mechanism(s) of the vaccine in the outbred CD-1 mice-S. mansoni model. Mice were percutaneously exposed to S. mansoni cercariae following immunization twice with 0 or 10 μg S. mansoni recombinant cathepsin B1 (SmCB1) or L3 (SmCL3). They were examined at specified intervals post infection (pi) for the level of serum antibodies, uric acid, which amplifies type 2 immune responses and is an anti-oxidant, lipids, in particular, arachidonic acid (ARA), which is an endoschistosomicide and ovocide, as well as uric acid and ARA in the lung and liver. Memory IgG1, IgG2a, and IgG2b antibodies to the cysteine peptidase immunogen were detectable at and following day 17 pi. Serum, lung, and liver uric acid levels in immunized mice were higher than in naïve and unimmunized mice, likely as a consequence of cysteine peptidase-mediated catabolic activity. Increased circulating uric acid in cysteine peptidase-immunized mice was associated with elevation in the amount of ARA in lung and liver at every test interval, and in serum starting at day 17 pi. Together, the results suggest the collaboration of humoral antibodies and ARA schistosomicidal potential in the attrition of challenge S. mansoni (p < 0.0005) at the liver stage, and ARA direct parasite egg killing (p < 0.005). The anti-oxidant and reactive oxygen species-scavenger properties of uric acid may be responsible for the cysteine peptidase vaccine protection ceiling. This article represents a step towards clarifying the protection mechanism of the cysteine peptidase-based schistosomiasis vaccine.

Introduction: Vaccination of hamsters with Schistosoma mansoni adjuvant-free recombinant cathepsin B1 (SmCB1) and L3 (SmCL3) have been shown to elicit highly significant (P < 0.005) protection against challenge Schistosoma haematobium that was not very superior to that achieved by the cysteine peptidase, papain. Sterilizing immunity might, however, be induced if hamsters were vaccinated against S. haematobium infection with a homologous cysteine peptidase, i.e., S. haematobium cathepsin L (ShCL). Methods: Standards methods, techniques, and primers based on the published nucleotide sequence of ShCL were used to clone, amplify and express DNAs encoding the target enzyme in a bacterial expression vector. Repeat immunization trials were performed using recombinant ShCL alone or in combination with the vaccine candidate S. mansoni recombinant glyceraldehyde 3-phosphate dehydrogenase, in parallel with S. mansoni lecucine aminopeptidase. Results: The results together indicated that our adjuvant-free, cysteine peptidase-based vaccine elicits highly significant (P < 0.0001) reduction in challenge worm burden and parasite egg viability. Protection was associated with whole blood cultures release of type 1, type 2, and type 17 cytokines, and modest, yet significant (P < 0.05) humoral response to ShCL. Conclusion: Sterilizing immunity was, however, not achieved in any trial, likely because of the preponderant role of cysteine peptidasesinduced nonspecific factors in amplifying and antagonizing its protective potential. Experiments are planned in an aim to identify these elusive factors and their exact role.