Extracellular Onchocerca-derived small RNAs in host nodules and blood

Particle and Fibre Toxicology

Amy H Buck⁷  Achim Hoerauf¹  David W Taylor⁵  Vincent N Tanya⁶  Germanus S Bah⁴  Henrietta F Ngangyung⁴  Samuel Wanji³  Alexander Debrah⁸  Mark Blaxter⁷  Kenneth M Pfarr¹  Alasdair Ivens⁷  Simon A Babayan²  Benjamin L Makepeace⁹  Juan F Quintana⁷ 

[1] Institute of Medical Microbiology, Immunology and Parasitology, University Hospital of Bonn, Bonn, Germany;Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK;Research Foundation in Tropical Diseases and Environment and University of Buea, Buea, Cameroon;Institut de Recherche Agricole pour le Développement, Regional Centre of Wakwa, Ngaoundéré, Cameroon;Division of Pathway Medicine, School for Biomedical Sciences, University of Edinburgh, Little France, Edinburgh, UK;Cameroon Academy of Sciences, Yaoundé, Cameroon;Centre for Immunity, Infection and Evolution, Ashworth Laboratories, University of Edinburgh, West Mains Road, Edinburgh, UK;Kumasi Centre for Collaborative Research (KCCR) and Kwame Nkrumah University of Science and Technology, Kumasi, Ghana;Institute of Infection and Global Health, University of Liverpool, Liverpool, Merseyside, UK

关键词: Host-pathogen;    Onchocerciasis;    Filarial nematode;    Extracellular RNA;    microRNAs;   

Others  :  1147322
DOI  :  10.1186/s13071-015-0656-1

 received in 2014-12-05, accepted in 2015-01-12,  发布年份 2015

【 摘 要 】

Background

microRNAs (miRNAs), a class of short, non-coding RNA can be found in a highly stable, cell-free form in mammalian body fluids. Specific miRNAs are secreted by parasitic nematodes in exosomes and have been detected in the serum of murine and dog hosts infected with the filarial nematodes Litomosoides sigmodontis and Dirofilaria immitis, respectively. Here we identify extracellular, parasite-derived small RNAs associated with Onchocerca species infecting cattle and humans.

Methods

Small RNA libraries were prepared from total RNA extracted from the nodule fluid of cattle infected with Onchocerca ochengi as well as serum and plasma from humans infected with Onchocerca volvulus in Cameroon and Ghana. Parasite-derived miRNAs were identified based on the criteria that sequences unambiguously map to hairpin structures in Onchocerca genomes, do not align to the human genome and are not present in European control serum.

Results

A total of 62 mature miRNAs from 52 distinct pre-miRNA candidates were identified in nodule fluid from cattle infected with O. ochengi of which 59 are identical in the genome of the human parasite O. volvulus. Six of the extracellular miRNAs were also identified in sequencing analyses of serum and plasma from humans infected with O. volvulus. Based on sequencing analysis the abundance levels of the parasite miRNAs in serum or plasma range from 5 to 127 reads/per million total host miRNA reads identified, comparable to our previous analyses of Schistosoma mansoni and L. sigmodontis miRNAs in serum. All six of the O. volvulus miRNAs identified have orthologs in other filarial nematodes and four were identified in the serum of mice infected with L. sigmodontis.

Conclusions

We have identified parasite-derived miRNAs associated with onchocerciasis in cattle and humans. Our results confirm the conserved nature of RNA secretion by diverse nematodes. Additional species-specific small RNAs from O. volvulus may be present in serum based on the novel miRNA sequences identified in the nodule fluid. In our analyses comparison to European control serum illuminates the scope for false-positives, warranting caution in criteria that should be applied to identification of biomarkers of infection.

【 授权许可】

   
2015 Quintana et al.; licensee BioMed Central.

【 图 表 】

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