Multiple sequence alignment of PPTSP2.5 and the SP2.5-like proteins from <i>Phlebotomus duboscqi</i> (Pd), <i>Phlebotomus perniciosus</i> (Pr), <i>Phlebotomus tobbi</i> (Pt) and <i>Phlebotomus arabicus</i> (Pa).

<p>Underlining indicates the predicted signal peptide sequence; black shading represents identity between amino acids of the predicted mature peptide.</p

Phylogenetic analysis of the small OBP family of proteins from <i>Phlebotomus papatasi</i> Israeli strain (Pp), <i>P. duboscqi</i> (Pd), <i>P. sergenti</i> (Ps), <i>P. tobbi</i> (Pt), <i>P. perniciosus</i> (Pp), <i>P. ariasi</i> (Pa), <i>P. arabicus</i> (Pa), <i>P. argentipes</i> (Pa), and <i>Lutzomyia longipalpis</i> (Ll), <i>Aedes aegypti</i> (Aa), and <i>Anopheles gambiae</i> (Ag).

<p>Accession numbers are next to each species designation, and node values indicate branch support.</p

Sequence alignment of PPTSP15 with the PPSP15 from <i>Phlebotomus papatasi</i> Israeli strain (PpAAL11047).

<p>Black shading represents identical amino acids.</p

Alignment of Lufaxin, the Factor Xa inhibitor from the saliva of <i>Lutzomyia longipalpis</i>, and the PPTSP34 salivary protein from <i>Phlebotomus papatasi</i> Tunisian strain.

<p>Black background shading represents identical amino acids.</p

Updating the Salivary Gland Transcriptome of <em>Phlebotomus papatasi</em> (Tunisian Strain): The Search for Sand Fly-Secreted Immunogenic Proteins for Humans

<div><h3>Introduction</h3><p>Sand fly saliva plays an important role in both blood feeding and outcome of <em>Leishmania</em> infection. A cellular immune response against a <em>Phlebotomus papatasi</em> salivary protein was shown to protect rodents against <em>Leishmania major</em> infection. In humans, <em>P. papatasi</em> salivary proteins induce a systemic cellular immune response as well as a specific antisaliva humoral immune response, making these salivary proteins attractive targets as markers of exposure for this <em>Leishmania</em> vector. Surprisingly, the repertoire of salivary proteins reported for <em>P. papatasi</em>–a model sand fly for <em>Leishmania</em>-vector-host molecular interactions–is very limited compared with other sand fly species. We hypothesize that a more comprehensive study of the transcripts present in the salivary glands of <em>P. papatasi</em> will provide better knowledge of the repertoire of proteins of this important vector and will aid in selection of potential immunogenic proteins for humans and of those proteins that are highly conserved between different sand fly strains.</p> <h3>Methods and Findings</h3><p>A cDNA library from <em>P. papatasi</em> (Tunisian strain) salivary glands was constructed, and randomly selected transcripts were sequenced and analyzed. The most abundant transcripts encoding secreted proteins were identified and compared with previously reported sequences. Importantly, we identified salivary proteins not described before in this sand fly species.</p> <h3>Conclusions</h3><p>Comparative analysis between the salivary proteins of <em>P. papatasi</em> from Tunisia and Israel strains shows a high level of identity, suggesting these proteins as potential common targets for markers of vector exposure or inducers of cellular immune responses in humans for different geographic areas.</p> </div

Multiple sequence alignment of the large OBP (D7 related proteins PPTSP30 and PPTSP28) and the small molecular weight OBP from the salivary glands of <i>Phlebotomus papatasi</i>.

<p>Multiple sequence alignment of the large OBP (D7 related proteins PPTSP30 and PPTSP28) and the small molecular weight OBP from the salivary glands of <i>Phlebotomus papatasi</i>.</p

Multiple sequence alignment of the two distinct members of the PPTSP14.2 family of proteins from the saliva of <i>Phlebotomus papatasi</i>.

<p>Black shading represents identical amino acids.</p

Alignment of <i>Anopheles stephensi</i> D7 protein (ANST D7L1) and the sand fly D7 protein (PPTSP28a) from <i>Phlebotomus papatasi</i> Tunisian strain.

<p>Black background shading represents identical amino acids. Yellow background shading represents amino acids absent in the sand fly D7 compared with the An. stephensi D7 protein.</p

Multiple sequence alignment of yellow-related proteins from <i>Phlebotomus papatasi</i> PPTSP42 and PPTSP44 and two yellow proteins from <i>Lutzomyia longipalpis</i> LJM11 and LJM17.

<p>Black shading represents identity. *Indicates essential amino acids for LJM11 and LJM17 binding of biogenic amines.</p

Phylogenetic analysis of sand fly salivary proteins of two D7 salivary proteins identified in this cDNA library (PPTSP28 and PPTSP30) and the D7 salivary proteins from <i>Phlebotomus papatasi</i> Israeli strain, <i>P. duboscqi, P. sergenti, P. tobbi, P. perniciosus, P. ariasi, P. arabicus, Lutzomyia longipalpis</i>, and <i>P. argentipes</i>, and other Diptera including <i>Drosophila melanogaster, Glossina morsitans morsitans, Culex quinquefasciatus</i>, and <i>Anopheles gambiae</i>.

<p>Accession numbers are next to each species, and node values indicate branch support.</p