Delivery of a functional anti-trypanosome Nanobody in different tsetse fly tissues via a bacterial symbiont, Sodalis glossinidius
Delivery of a functional anti-trypanosome Nanobody in different tsetse fly tissues via a bacterial symbiont, Sodalis glossinidius
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DOI：
10.1186/s12934-014-0156-6
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发表时间：
2014-11-07
影响因子：
6.4
通讯作者：
Van den Abbeele, Jan
中科院分区：
文献类型：
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作者：
De Vooght, Linda;Caljon, Guy;Van den Abbeele, Jan
背景：Sodalis folsinidius 是采采蝇垂直传播的微生物共生体，目前被认为是抗锥虫成分的潜在递送系统，可减少或消除采采蝇宿主传播寄生锥虫的能力，这种方法也称为副转基因。开发副转基因采采蝇的一个重要步骤是用重组 Sodalis 细菌在成年果蝇及其后代中稳定定殖，在寄生虫所在的组织中表达杀锥虫效应分子。 结果：在这项研究中，测试了 Sodalis 在 Glossina morsitans morsitans 中传递功能性抗锥虫纳米抗体 (Nbs) 的能力。我们表征了表达强效锥虫纳米抗体（即 Nb_An46）的重组 Sodalis (recSodalis) 的体外和体内稳定性。我们表明，recSodalis 在体内条件下与 WT Sodalis 具有竞争性，并且短暂清除其内源 WT Sodalis 种群的采采蝇可以成功地用高密度的 recSodalis 重新繁殖。此外，还观察到向后代的垂直传播。最后，我们证明了recSodalis在不同的采采蝇组织中表达了显着水平（ng范围）的功能性Nb_An46，包括锥虫寄生虫发生重要发育阶段的中肠。结论：我们证明了Sodalis共生体可以通过基因工程在采采蝇的不同组织中表达和释放大量的功能性抗锥虫Nbs。这种利用昆虫共生细菌传递的病原体靶向纳米抗体的创新概念的应用可以扩展到其他载体病原体系统。
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.