Engineered vaginal Lactobacillus strain for mucosal delivery of the human immunodeficiency virus inhibitor cyanovirin-N
Engineered vaginal Lactobacillus strain for mucosal delivery of the human immunodeficiency virus inhibitor cyanovirin-N
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DOI：
10.1128/aac.00493-06
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发表时间：
2006-10-01
影响因子：
4.9
通讯作者：
Xu, Qiang
中科院分区：
文献类型：
--
作者：
Liu, Xiaowen;Lagenaur, Laurel A.;Xu, Qiang
妇女感染人类免疫缺陷病毒(HIV)的风险很大，宫颈阴道粘膜是病毒进入的主要门户。迫切需要由女性发起的预防措施，包括局部杀微生物剂，以帮助遏制艾滋病毒/艾滋病的流行。在这里，我们报告了一种新型的、活的杀微生物剂的开发，它使用了一种天然的詹氏乳杆菌阴道菌株，该菌株被设计成能够输送有效的艾滋病毒抑制剂氰基韦林N(CV-N)。为了促进这种细菌高效表达CV-N，对L.jensenii 1153基因组进行了测序，从而鉴定了异源基因的染色体整合的天然调控元件和位置。对CV-N表达盒进行了优化，结果表明，在简氏乳杆菌中表达的CV-N具有较高的结构完整性。乳杆菌来源的CV-N在体外能够抑制CCR5嗜性HIVBaL的感染性，其50%抑制浓度为0.3 nM。Cv-N表达盒稳定地以单拷贝形式整合到细菌染色体中，并从外源质粒DNA中分离出来，而不会对细菌的表型产生不利影响。这种细菌能够在发情期在小鼠阴道内定植并产生全长的CV-N。产生CV-N的乳酸菌在体外和体内繁殖时遗传稳定。这项工作代表着朝着开发一种廉价但耐用的基于蛋白质的杀微生物剂迈出了重要的一步，该杀菌剂可以阻止艾滋病毒在妇女中的异性传播。
Women are at significant risk of human immunodeficiency virus (HIV) infection, with the cervicovaginal mucosa serving as a major portal for virus entry. Female-initiated preventatives, including topical microbicides, are urgently needed to help curtail the HIV/AIDS pandemic. Here we report on the development of a novel, live microbicide that employs a natural vaginal strain of Lactobacillus jensenii engineered to deliver the potent HIV inhibitor cyanovirin-N (CV-N). To facilitate efficient expression of CV-N by this bacterium, the L. jensenii 1153 genome was sequenced, allowing identification of native regulatory elements and sites for the chromosomal integration of heterologous genes. A CV-N expression cassette was optimized and shown to produce high levels of structurally intact CV-N when expressed in L. jensenii. Lactobacillus-derived CV-N was capable of inhibiting CCR5-tropic HIVBaL infectivity in vitro with a 50% inhibitory concentration of 0.3 nM. The CV-N expression cassette was stably integrated as a single copy into the bacterial chromosome and resolved from extraneous plasmid DNA without adversely affecting the bacterial phenotype. This bacterial strain was capable of colonizing the vagina and producing full-length CV-N when administered intravaginally to mice during estrus phase. The CV-N-producing Lactobacillus was genetically stable when propagated in vitro and in vivo. This work represents a major step towards the development of an inexpensive yet durable protein-based microbicide to block the heterosexual transmission of HIV in women.