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Delivery of a functional anti-trypanosome nanobody in different tsetse fly tissues via a bacterial symbiont, Sodalis glossinidius

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Abstract
Background: Sodalis glossinidius, a vertically transmitted microbial symbiont of the tsetse fly, is currently considered as a potential delivery system for anti-trypanosomal components that reduce or eliminate the capability of the tsetse fly host to transmit parasitic trypanosomes, an approach also known as paratransgenesis. An essential step in developing paratransgenic tsetse is the stable colonization of adult flies and their progeny with recombinant Sodalis bacteria, expressing trypanocidal effector molecules in tissues where the parasite resides. Results: In this study, Sodalis was tested for its ability to deliver functional anti-trypanosome nanobodies (Nbs) in Glossina morsitans morsitans. We characterized the in vitro and in vivo stability of recombinant Sodalis (recSodalis) expressing a potent trypanolytic nanobody, i.e. Nb_An46. We show that recSodalis is competitive with WT Sodalis in in vivo conditions and that tsetse flies transiently cleared of their endogenous WT Sodalis population can be successfully repopulated with recSodalis at high densities. In addition, vertical transmission to the offspring was observed. Finally, we demonstrated that recSodalis expressed significant levels (ng range) of functional Nb_An46 in different tsetse fly tissues, including the midgut where an important developmental stage of the trypanosome parasite occurs. Conclusions: We demonstrated the proof-of-concept that the Sodalis symbiont can be genetically engineered to express and release significant amounts of functional anti-trypanosome Nbs in different tissues of the tsetse fly. The application of this innovative concept of using pathogen-targeting nanobodies delivered by insect symbiotic bacteria could be extended to other vector-pathogen systems.
Keywords
Sodalis glossinidius, (3-10), Symbiont, Paratransgenesis, Recombinant, Glossina, Delivery, Functional, Nanobody, In vivo, Midgut, EXPRESSION, MOSQUITOS, MALARIA

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Citation

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MLA
De Vooght, Linda et al. “Delivery of a Functional Anti-trypanosome Nanobody in Different Tsetse Fly Tissues via a Bacterial Symbiont, Sodalis Glossinidius.” MICROBIAL CELL FACTORIES 13 (2014): n. pag. Print.
APA
De Vooght, L., Caljon, G., De Ridder, K., & Van Den Abbeele, J. (2014). Delivery of a functional anti-trypanosome nanobody in different tsetse fly tissues via a bacterial symbiont, Sodalis glossinidius. MICROBIAL CELL FACTORIES, 13.
Chicago author-date
De Vooght, Linda, Guy Caljon, Karina De Ridder, and Jan Van Den Abbeele. 2014. “Delivery of a Functional Anti-trypanosome Nanobody in Different Tsetse Fly Tissues via a Bacterial Symbiont, Sodalis Glossinidius.” Microbial Cell Factories 13.
Chicago author-date (all authors)
De Vooght, Linda, Guy Caljon, Karina De Ridder, and Jan Van Den Abbeele. 2014. “Delivery of a Functional Anti-trypanosome Nanobody in Different Tsetse Fly Tissues via a Bacterial Symbiont, Sodalis Glossinidius.” Microbial Cell Factories 13.
Vancouver
1.
De Vooght L, Caljon G, De Ridder K, Van Den Abbeele J. Delivery of a functional anti-trypanosome nanobody in different tsetse fly tissues via a bacterial symbiont, Sodalis glossinidius. MICROBIAL CELL FACTORIES. 2014;13.
IEEE
[1]
L. De Vooght, G. Caljon, K. De Ridder, and J. Van Den Abbeele, “Delivery of a functional anti-trypanosome nanobody in different tsetse fly tissues via a bacterial symbiont, Sodalis glossinidius,” MICROBIAL CELL FACTORIES, vol. 13, 2014.
@article{6861071,
  abstract     = {Background: Sodalis glossinidius, a vertically transmitted microbial symbiont of the tsetse fly, is currently considered as a potential delivery system for anti-trypanosomal components that reduce or eliminate the capability of the tsetse fly host to transmit parasitic trypanosomes, an approach also known as paratransgenesis. An essential step in developing paratransgenic tsetse is the stable colonization of adult flies and their progeny with recombinant Sodalis bacteria, expressing trypanocidal effector molecules in tissues where the parasite resides. 
Results: In this study, Sodalis was tested for its ability to deliver functional anti-trypanosome nanobodies (Nbs) in Glossina morsitans morsitans. We characterized the in vitro and in vivo stability of recombinant Sodalis (recSodalis) expressing a potent trypanolytic nanobody, i.e. Nb_An46. We show that recSodalis is competitive with WT Sodalis in in vivo conditions and that tsetse flies transiently cleared of their endogenous WT Sodalis population can be successfully repopulated with recSodalis at high densities. In addition, vertical transmission to the offspring was observed. Finally, we demonstrated that recSodalis expressed significant levels (ng range) of functional Nb_An46 in different tsetse fly tissues, including the midgut where an important developmental stage of the trypanosome parasite occurs. 
Conclusions: We demonstrated the proof-of-concept that the Sodalis symbiont can be genetically engineered to express and release significant amounts of functional anti-trypanosome Nbs in different tissues of the tsetse fly. The application of this innovative concept of using pathogen-targeting nanobodies delivered by insect symbiotic bacteria could be extended to other vector-pathogen systems.},
  articleno    = {156},
  author       = {De Vooght, Linda and Caljon, Guy and De Ridder, Karina and Van Den Abbeele, Jan},
  issn         = {1475-2859},
  journal      = {MICROBIAL CELL FACTORIES},
  keywords     = {Sodalis glossinidius,(3-10),Symbiont,Paratransgenesis,Recombinant,Glossina,Delivery,Functional,Nanobody,In vivo,Midgut,EXPRESSION,MOSQUITOS,MALARIA},
  language     = {eng},
  pages        = {10},
  title        = {Delivery of a functional anti-trypanosome nanobody in different tsetse fly tissues via a bacterial symbiont, Sodalis glossinidius},
  url          = {http://dx.doi.org/10.1186/s12934-014-0156-6},
  volume       = {13},
  year         = {2014},
}

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