Structure of SALO, a leishmaniasis vaccine candidate from the sand fly Lutzomyia longipalpis.
BACKGROUND:Immunity 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 protecti...
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doaj-79f22f7975c54654901cb554512a58862020-11-24T21:41:57ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352017-03-01113e000537410.1371/journal.pntd.0005374Structure of SALO, a leishmaniasis vaccine candidate from the sand fly Lutzomyia longipalpis.Oluwatoyin A AsojoAlan KelleherZhuyun LiuJeroen PolletElissa M HudspethWanderson C RezendeMallory Jo GroenChristopher A SeidMaha AbdeladhimShannon TownsendWaldione de CastroAntonio Mendes-SousaDaniella Castanheira BartholomeuRicardo Toshio FujiwaraMaria Elena BottazziPeter J HotezBin ZhanFabiano OliveiraShaden KamhawiJesus G ValenzuelaBACKGROUND:Immunity 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 FINDINGS:Recombinant 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/SIGNIFICANCE:The 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.http://europepmc.org/articles/PMC5344329?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Oluwatoyin A Asojo Alan Kelleher Zhuyun Liu Jeroen Pollet Elissa M Hudspeth Wanderson C Rezende Mallory Jo Groen Christopher A Seid Maha Abdeladhim Shannon Townsend Waldione de Castro Antonio Mendes-Sousa Daniella Castanheira Bartholomeu Ricardo Toshio Fujiwara Maria Elena Bottazzi Peter J Hotez Bin Zhan Fabiano Oliveira Shaden Kamhawi Jesus G Valenzuela |
spellingShingle |
Oluwatoyin A Asojo Alan Kelleher Zhuyun Liu Jeroen Pollet Elissa M Hudspeth Wanderson C Rezende Mallory Jo Groen Christopher A Seid Maha Abdeladhim Shannon Townsend Waldione de Castro Antonio Mendes-Sousa Daniella Castanheira Bartholomeu Ricardo Toshio Fujiwara Maria Elena Bottazzi Peter J Hotez Bin Zhan Fabiano Oliveira Shaden Kamhawi Jesus G Valenzuela Structure of SALO, a leishmaniasis vaccine candidate from the sand fly Lutzomyia longipalpis. PLoS Neglected Tropical Diseases |
author_facet |
Oluwatoyin A Asojo Alan Kelleher Zhuyun Liu Jeroen Pollet Elissa M Hudspeth Wanderson C Rezende Mallory Jo Groen Christopher A Seid Maha Abdeladhim Shannon Townsend Waldione de Castro Antonio Mendes-Sousa Daniella Castanheira Bartholomeu Ricardo Toshio Fujiwara Maria Elena Bottazzi Peter J Hotez Bin Zhan Fabiano Oliveira Shaden Kamhawi Jesus G Valenzuela |
author_sort |
Oluwatoyin A Asojo |
title |
Structure of SALO, a leishmaniasis vaccine candidate from the sand fly Lutzomyia longipalpis. |
title_short |
Structure of SALO, a leishmaniasis vaccine candidate from the sand fly Lutzomyia longipalpis. |
title_full |
Structure of SALO, a leishmaniasis vaccine candidate from the sand fly Lutzomyia longipalpis. |
title_fullStr |
Structure of SALO, a leishmaniasis vaccine candidate from the sand fly Lutzomyia longipalpis. |
title_full_unstemmed |
Structure of SALO, a leishmaniasis vaccine candidate from the sand fly Lutzomyia longipalpis. |
title_sort |
structure of salo, a leishmaniasis vaccine candidate from the sand fly lutzomyia longipalpis. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS Neglected Tropical Diseases |
issn |
1935-2727 1935-2735 |
publishDate |
2017-03-01 |
description |
BACKGROUND:Immunity 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 FINDINGS:Recombinant 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/SIGNIFICANCE:The 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. |
url |
http://europepmc.org/articles/PMC5344329?pdf=render |
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