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Onchocerca-derived small RNAs in host nodules and serum

ABSTRACT: We report the presence of Onchocerca ochengi and O. volvulus derived small RNAs in bovine nodule fluids and human serum and plasma, respectively. Further comparisons with other related filarial nematodes like Litomosoides sigmodontis and Dirofilaria immitis reveal common and distictive signatures associated to the Onchocerca species.

ORGANISM(S): Bos taurus Homo sapiens

PROVIDER: GSE63933 | GEO | 2015/04/24

SECONDARY ACCESSION(S): PRJNA269539

REPOSITORIES: GEO

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Project description:Filarial nematodes (superfamily Filarioidea) are responsible for an annual global health burden of approximately 6.3 million disability-adjusted life-years, which represents the greatest single component of morbidity attributable to helminths affecting humans. No vaccine exists for the major filarial diseases, lymphatic filariasis and onchocerciasis; in part because research on protective immunity against filariae has been constrained because the human-parasitic species cannot complete their lifecycles in laboratory mice. However, the rodent filaria Litomosoides sigmodontis has become a popular experimental model, as BALB/c mice are fully permissive for its development and reproduction. Here, we provide a comprehensive analysis of excretory-secretory products from L. sigmodontis across five lifecycle stages. Applying intensity-based quantification, we determined the abundance of 302 unique excretory-secretory proteins, of which 64.6% were present in quantifiable amounts only from gravid adult female nematodes. This lifecycle stage, together with immature first-stage larvae (microfilariae), released four proteins that have not previously been evaluated as vaccine candidates: a predicted 28.5 kDa filaria-specific protein, a zonadhesin and SCO-spondin-like protein, a vitellogenin, and a protein containing six metridin-like ShK toxin domains. Female nematodes also released two proteins derived from the obligate Wolbachia symbiont. Notably, excretory-secretory products from all parasite stages contained several uncharacterised members of the transthyretin-like protein family. Furthermore, biotin labelling revealed that redox proteins and enzymes involved in purinergic signalling were enriched on the adult nematode cuticle. Comparison of the L. sigmodontis adult secretome with that of the human–infective filarial nematode Brugia malayi (reported previously in three independent published studies) identified differences that suggest a considerable underlying diversity of potential immunomodulators. The molecules identified in L. sigmodontis excretory-secretory products show promise not only for vaccination against filarial infections, but for the amelioration of allergy and autoimmune diseases.

Project description:The zoonotic mosquito-borne filarial nematode Dirofilaria repens causes subcutaneous and ocular infections in dogs, cats and humans. Microfilariae (mf) are taken up by mosquitoes from infected vertebrate hosts which develop in the mosquito to the infectious third stage larvae (L3). These are transmitted to new vertebrate hosts and develop over two further moults to adult worms. The aims of the project were 1) the de novo sequencing and annotation of the D. repens genome and 2) comparative transcriptome analyses of the developmental stages mf and L3 as we hypothesized remarkable differences in gene expression between the different developmental stages. Genomic DNA was obtained from adult male D. repens. RNA was extracted from microfilariae from naturally infected dogs and from L3 produced in Aedes aegypti mosquitoes fed on blood spiked with microfi-lariae. The 99.59 MB genome was around 17% larger than that of the related species D. immitis (dog heartworm) and contained 8.9% fewer predicted genes (10,357). Around 1.8% of identified proteins (206/11,262) could not be mapped to D. immitis. Out of these, 6 (2.9 %) presented an ortholog in all other considered filarial nematodes (e.g. Loa loa) and Caenorhabditis elegans. A sig-nificantly higher number of D. repens proteins as compared to D. immitis mapped to the filarial nem-atode L. loa, reflecting the similarity in biology of D. repens and L. loa. A total of 876 genes were differentially expressed, of which 591 could be annotated in UniProtKB/Swiss-Prot. In particular, 155 genes with a UniProtKB/Swiss-Prot annotation to C. elegans and filarial nematodes were upregulat-ed in the L3 and 57 in the mf stage, respectively. Fifteen GO BP (Gene Ontology Biological Pro-cesses) were significantly enriched for the L3 group and 12 for the mf. These data provide first in-sight into the differential gene expression profiles of this filarial nematode and can serve for future investigations of metabolic processes and stage-specific diagnostics.

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Onchocerca-derived small RNAs in host nodules and serum

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