SALO, a novel classical pathway complement inhibitor from saliva of the sand fly Lutzomyia longipalpis

Ferreira, Viviana P.; Vale, Vladimir Fazito do; Pangburn, Michael K.; Abdeladhim, Maha; Mendes-Sousa, Antonio Ferreira; Coutinho-Abreu, Iliano V.; Rasouli, Manoochehr; Brandt, Ernst; Meneses, Claudio; Lima, Kolyvan Ferreira; Araújo, Ricardo Nascimento; Pereira, Marcos H.; Kotsyfakis, Michail; Oliveira, Fabiano; Kamhawi, Shaden; Ribeiro, José M. C.; Gontijo, Nelder F.; Collin, Nicolas; Valenzuela, Jesús G.

doi:10.1038/srep19300

Cited by 41 publications

(47 citation statements)

References 42 publications

(51 reference statements)

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“…Anti-Factor B #1379, an inhibitory monoclonal antibody (41), and SALO, a salivary protein produced by the sandfly Lutzomyia longipalis (54-56) that specifically inhibits the CP (42), were used to determine the effects of AP and CP inhibition on TRAP-mediated PGA formation, respectively. Addition of either anti-Factor B #1379 or SALO to whole blood significantly impaired TRAP-mediated PGA formation in each donor’s blood by an average of ~50% (Figures 3A-B and 3C-D respectively, and Figure 4A-B), as well as C3 fragment deposition (Figure 4C-D) in a mode similar to the anti-properdin monoclonals (Figures 2 and 4).…”

Section: Resultsmentioning

confidence: 99%

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Properdin-Mediated C5a Production Enhances Stable Binding of Platelets to Granulocytes in Human Whole Blood

Blatt

Saggu

Kulkarni

et al. 2016

The Journal of Immunology

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Enhanced levels of platelet/granulocyte aggregates (PGAs) are found in patients suffering from many different inflammatory vascular diseases, and their formation in animal models of vascular disease is associated with increased thromboinflammation and worsened outcomes. The complement system, a part of the innate immune system, influences PGA formation, however the mechanisms for its effects are unknown. Here, we have defined complement-mediated mechanisms that enhance PGA formation in human whole blood stimulated with thrombin receptor activating peptide (TRAP) using ex-vivo flow cytometry assays. We demonstrate that physiological properdin, a positive regulator of complement alternative pathway activity, increases PGA formation when added to TRAP-stimulated blood. All physiological properdin forms increase PGA formation, but properdin tetramers are the most efficient at increasing complement activity and PGA formation. Inhibition of endogenous properdin, either circulating in the blood or produced locally by leukocytes, impairs TRAP-mediated PGA formation to the same level as specific inhibition of either the alternative or classical pathways. In addition, blocking the interaction of C5a with its cellular receptor prevents properdin-mediated increases in PGA formation. Adding either properdin tetramers or C5a to whole blood increases CD11b expression on granulocytes and this increase is prevented by blockade of the C5a-C5a receptor axis. Finally, we demonstrate that the effects of properdin on PGA formation are tightly regulated by Factor H. Cumulatively, our data indicate that properdin enhances PGA formation via increased production of C5a, and that inhibition of properdin function has therapeutic potential to limit thromboinflammation in diseases characterized by increased PGA formation.

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Section: Resultsmentioning

confidence: 99%

“…1G6D2, which does not inhibit properdin function, was used as both a negative inhibition control and isotype control. SALO, a specific CP inhibitor, was generated as previously described (42). C5a was purchased from Complement Technologies.…”

Section: Methodsmentioning

confidence: 99%

Properdin-Mediated C5a Production Enhances Stable Binding of Platelets to Granulocytes in Human Whole Blood

Blatt

Saggu

Kulkarni

et al. 2016

The Journal of Immunology

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“…Furthermore, this vaccine candidate was recently shown to inhibit the classical pathway of complement [10]. To move this vaccine candidate towards the clinical path, we solved the structure of SALO and expressed it in P .…”

Section: Discussionmentioning

confidence: 99%

Structure of SALO, a leishmaniasis vaccine candidate from the sand fly Lutzomyia longipalpis

et al. 2017

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BackgroundImmunity to the sand fly salivary protein SALO (Salivary Anticomplement of Lutzomyia longipalpis) protected hamsters against Leishmania infantum and L. braziliensis infection and, more recently, a vaccine combination of a genetically modified Leishmania with SALO conferred strong protection against L. donovani infection. Because of the importance of SALO as a potential component of a leishmaniasis vaccine, a plan to produce this recombinant protein for future scale manufacturing as well as knowledge of its structural characteristics are needed to move SALO forward for the clinical path.Methodology/Principal findingsRecombinant SALO was expressed as a soluble secreted protein using Pichia pastoris, rSALO(P), with yields of 1g/L and >99% purity as assessed by SEC-MALS and SDS-PAGE. Unlike its native counterpart, rSALO(P) does not inhibit the classical pathway of complement; however, antibodies to rSALO(P) inhibit the anti-complement activity of sand fly salivary gland homogenate. Immunization with rSALO(P) produces a delayed type hypersensitivity response in C57BL/6 mice, suggesting rSALO(P) lacked anti-complement activity but retained its immunogenicity. The structure of rSALO(P) was solved by S-SAD at Cu-Kalpha to 1.94 Å and refined to Rfactor 17%. SALO is ~80% helical, has no appreciable structural similarities to any human protein, and has limited structural similarity in the C-terminus to members of insect odorant binding proteins. SALO has three predicted human CD4+ T cell epitopes on surface exposed helices.Conclusions/SignificanceThe results indicate that SALO as expressed and purified from P. pastoris is suitable for further scale-up, manufacturing, and testing. SALO has a novel structure, is not similar to any human proteins, is immunogenic in rodents, and does not have the anti-complement activity observed in the native salivary protein which are all important attributes to move this vaccine candidate forward to the clinical path.

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“…Salivary proteins or peptides previously identified exclusively in transcriptomes of NW sand flies include vasodilatory peptide maxadilan [61], salivary 5 0 -nucleotidase, an enzyme responsible for cleaving AMP to adenosine [62], SALO anti-complement proteins [63], the Table 3. Salivary apyrase and hyaluronidase in two S. schwetzi lineages maintained on different blood-meal sources, geckos (S-G) and mice (S-M).…”

Section: Plos Onementioning

confidence: 99%

Sergentomyia schwetzi: Salivary gland transcriptome, proteome and enzymatic activities in two lineages adapted to different blood sources

et al. 2020

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During the blood feeding, sand fly females inject saliva containing immunomodulatory and anti-haemostatic molecules into their vertebrate hosts. The saliva composition is species-specific, likely due to an adaptation to particular haemostatic pathways of their preferred host. Research on sand fly saliva is limited to the representatives of two beststudied genera, Phlebotomus and Lutzomyia. Although the members of the genus Sergentomyia are highly abundant in many areas in the Old World, their role in human disease transmission remains uncertain. Most Sergentomyia spp. preferentially attack various species of reptiles, but feeding on warm-blooded vertebrates, including humans and domestic animals, has been repeatedly described, especially for Sergentomyia schwetzi, of which salivary gland transcriptome and proteome is analyzed in the current study. Illumina RNA sequencing and de novo assembly of the reads and their annotation revealed 17,293 sequences homologous to other arthropods' proteins. In the sialome, all proteins typical for sand fly saliva were identified-antigen 5-related, lufaxin, yellowrelated, PpSP15-like, D7-related, ParSP25-like, and silk proteins, as well as less frequent salivary proteins included 71kDa-like, ParSP80-like, SP16-like, and ParSP17-like proteins. Salivary enzymes include apyrase, hyaluronidase, endonuclease, amylase, lipase A2, adenosine deaminase, pyrophosphatase, 5'nucleotidase, and ribonuclease. Proteomics analysis of salivary glands identified 631 proteins, 81 of which are likely secreted into the saliva. We also compared two S. schwetzi lineages derived from the same origin. These lineages were adapted for over 40 generations for blood feeding either on mice (S-M) or geckos (S-G), two vertebrate hosts with different haemostatic mechanisms. Altogether, 20 and 40 annotated salivary transcripts were up-regulated in the S-M and S-G lineage, respectively. Proteomic comparison revealed ten salivary proteins more abundant in the lineage S-M, whereas 66 salivary proteins were enriched in the lineage S-G.

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SALO, a novel classical pathway complement inhibitor from saliva of the sand fly Lutzomyia longipalpis

Cited by 41 publications

References 42 publications

Properdin-Mediated C5a Production Enhances Stable Binding of Platelets to Granulocytes in Human Whole Blood

Properdin-Mediated C5a Production Enhances Stable Binding of Platelets to Granulocytes in Human Whole Blood

Structure of SALO, a leishmaniasis vaccine candidate from the sand fly Lutzomyia longipalpis

Sergentomyia schwetzi: Salivary gland transcriptome, proteome and enzymatic activities in two lineages adapted to different blood sources

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