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Open Access Research article

Analysis of salivary transcripts and antigens of the sand fly Phlebotomus arabicus

Jitka Hostomská1, Věra Volfová1, Jianbing Mu2, Mark Garfield3, Iva Rohoušová1, Petr Volf1, Jesus G Valenzuela2* and Ryan C Jochim2*

Author Affiliations

1 Charles University in Prague, Faculty of Science, Department of Parasitology, Vinicna 7, 128 44 Praha 2, Czech Republic

2 Vector Molecular Biology Unit, Laboratory of Malaria and Vector Research, NIAID, NIH, Rockville, MD, 20852, USA

3 Research Technologies Branch, NIAID, NIH, Rockville, MD, 20852, USA

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BMC Genomics 2009, 10:282  doi:10.1186/1471-2164-10-282

Published: 25 June 2009

Abstract

Background

Sand fly saliva plays an important role in blood feeding and Leishmania transmission as it was shown to increase parasite virulence. On the other hand, immunity to salivary components impedes the establishment of infection. Therefore, it is most desirable to gain a deeper insight into the composition of saliva in sand fly species which serve as vectors of various forms of leishmaniases. In the present work, we focused on Phlebotomus (Adlerius) arabicus, which was recently shown to transmit Leishmania tropica, the causative agent of cutaneous leishmaniasis in Israel.

Results

A cDNA library from salivary glands of P. arabicus females was constructed and transcripts were sequenced and analyzed. The most abundant protein families identified were SP15-like proteins, ParSP25-like proteins, D7-related proteins, yellow-related proteins, PpSP32-like proteins, antigen 5-related proteins, and 34 kDa-like proteins. Sequences coding for apyrases, hyaluronidase and other putative secreted enzymes were also represented, including endonuclease, phospholipase, pyrophosphatase, amylase and trehalase. Mass spectrometry analysis confirmed the presence of 20 proteins predicted to be secreted in the salivary proteome. Humoral response of mice bitten by P. arabicus to salivary antigens was assessed and many salivary proteins were determined to be antigenic.

Conclusion

This transcriptomic analysis of P. arabicus salivary glands is the first description of salivary proteins of a sand fly in the subgenus Adlerius. Proteomic analysis of P. arabicus salivary glands produced the most comprehensive account in a single sand fly species to date. Detailed information and phylogenetic relationships of the salivary proteins are provided, expanding the knowledge base of molecules that are likely important factors of sand fly-host and sand fly-Leishmania interactions. Enzymatic and immunological investigations further demonstrate the value of functional transcriptomics in advancing biological and epidemiological research that can impact leishmaniasis.