Deletion of Parasite Immune Modulatory Sequences Combined with Immune Activating Signals Enhances Vaccine Mediated Protection against Filarial Nematodes

Abstract

Filarial nematodes are tissue-dwelling parasites that can be killed by Th2-driven immune effectors, but that have evolved to withstand immune attack and establish chronic infections by suppressing host immunity. As a consequence, the efficacy of a vaccine against filariasis may depend on its capacity to counter parasite-driven immunomodulation. We immunised mice with DNA plasmids expressing functionally-inactivated forms of two immunomodulatory molecules expressed by the filarial parasite Litomosoides sigmodontis: the abundant larval transcript-1 (LsALT) and cysteine protease inhibitor-2 (LsCPI). The mutant proteins enhanced antibody and cytokine responses to live parasite challenge, and led to more leukocyte recruitment to the site of infection than their native forms. The immune response was further enhanced when the antigens were targeted to dendritic cells using a single chain Fv-αDEC205 antibody and co-administered with plasmids that enhance T helper 2 immunity (IL-4) and antigen-presenting cell recruitment (Flt3L, MIP-1α). Mice immunised simultaneously against the mutated forms of LsALT and LsCPI eliminated adult parasites faster and consistently reduced peripheral microfilaraemia. A multifactorial analysis of the immune response revealed that protection was strongly correlated with the production of parasite-specific IgG1 and with the numbers of leukocytes present at the site of infection. We have developed a successful strategy for DNA vaccination against a nematode infection that specifically targets parasite-driven immunosuppression while simultaneously enhancing Th2 immune responses and parasite antigen presentation by dendritic cells. Filarial infections are endemic in more that 80 countries, affecting over 120 million people and putting 1 billion more at risk. Antifilarial drugs must be administered regularly to infected people to control the disease, but they are contraindicated in under 6 year-olds and in pregnant women. Further, reports of drug resistance are now accumulating. A vaccine would therefore greatly help fight these diseases. Live attenuated L3 filariae larvae can evoke a protective immunity but their production is impractical and use in humans unacceptable while the efficacy of sub-unit vaccines has been poor. Filariae secrete proteins capable of suppressing their host's immune response, and have the potential to interfere with immunisation. We therefore decided to vaccinate hosts against secreted parasite products that modulate host immune responses rather than against structural components of the worms, and to boost the host's immune system by directly enhancing the uptake of parasite material by antigen presenting cells. This strategy generated substantial protection against both adult and offspring of a filarial parasite in mice. This provides a strong proof of principle for the anti-immunomodulatory approach we have developed.