Haloarchaeal gas vesicle nanoparticles displaying Salmonella SopB antigen reduce bacterial burden when administered with live attenuated bacteria.
Haloarchaeal gas vesicle nanoparticles displaying Salmonella SopB antigen reduce bacterial burden when administered with live attenuated bacteria.
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DOI：
10.1016/j.vaccine.2014.06.021
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发表时间：
2014-07-31
期刊：
影响因子：
5.5
通讯作者：
DasSarma S
中科院分区：
文献类型：
--
作者：
DasSarma P;Negi VD;Balakrishnan A;Karan R;Barnes S;Ekulona F;Chakravortty D;DasSarma S
迫切需要安全、有效且廉价的针对伤寒和其他沙门氏菌疾病的创新疫苗。为了满足这一需求，来自嗜盐古菌盐杆菌属的浮力、自我辅助的气体囊泡纳米粒子（GVNP）。 NRC-1 经过生物工程改造，可展示高度保守的肠沙门氏菌抗原 SopB，这是一种分泌型磷酸肌苷效应蛋白，由病原菌在感染宿主细胞期间注入。靠近 3' 编码区的两个高度保守的 sopB 基因片段，分别命名为 sopB4 和 B5，分别与 gvpC 基因的 C 端编码区融合，并通过离心加速浮选纯化所得 GVNP。使用针对 SopB 短合成肽产生的抗血清，通过蛋白质印迹分析建立了 SopB4 和 B5 抗原表位在 GVNP 上的展示。 SopB4 和 B5 纳米颗粒的免疫刺激活性通过向 BALB/c 小鼠腹腔内施用重组 GVNP 来测试，该小鼠已用减毒活疫苗株鼠伤寒沙门氏菌血清型 14028 ΔpmrG-HM-D (DV-STM-07) 免疫。在接受 SopB5-GVNP 强化的小鼠中，促炎细胞因子 IFN-γ、IL-2 和 IL-9 被显着诱导，这与强劲的 Th1 反应一致。在用有毒力的鼠伤寒沙门氏菌血清型 14028 攻击后，发现用 SopB4-GVNP 加强的小鼠脾脏中的细菌负荷减少，而用 SopB5-GVNP 加强的小鼠的肝脏、肠系膜淋巴结和脾中细菌负荷不存在或显着减少，这表明在 GVNP 上展示的 SopB 的 C 末端部分在小鼠中引发了对沙门氏菌感染的保护性反应。老鼠。发现 SopB 抗原-GVNP 在盐杆菌细胞中无需冷藏即可在高温下长时间保持稳定。所有结果都表明，生物工程 GVNP 可能成为开发针对沙门氏菌疾病的改良疫苗的宝贵平台。
Innovative vaccines against typhoid and other Salmonella diseases that are safe, effective, and inexpensive are urgently needed. In order to address this need, buoyant, self-adjuvating gas vesicle nanoparticles (GVNPs) from the halophilic archaeon Halobacterium sp. NRC-1 were bioengineered to display the highly conserved Salmonella enterica antigen SopB, a secreted inosine phosphate effector protein injected by pathogenic bacteria during infection into the host cell. Two highly conserved sopB gene segments near the 3’-coding region, named sopB4 and B5, were each fused to the C-terminal coding region of the gvpC gene, and resulting GVNPs were purified by centrifugally accelerated flotation. Display of SopB4 and B5 antigenic epitopes on GVNPs was established by Western blotting analysis using antisera raised against short synthetic peptides of SopB. Immunostimulatory activities of the SopB4 and B5 nanoparticles were tested by intraperitoneal administration of recombinant GVNPs to BALB/c mice which had been immunized with S. enterica serovar Typhimurium 14028 ΔpmrG-HM-D (DV-STM-07), a live attenuated vaccine strain. Proinflammatory cytokines IFN-γ, IL-2, and IL-9 were significantly induced in mice boosted with SopB5-GVNPs, consistent with a robust Th1 response. After challenge with virulent S. enterica serovar Typhimurium 14028, bacterial burden was found to be diminished in spleen of mice boosted with SopB4-GVNPs and absent or significantly diminished in liver, mesenteric lymph node, and spleen of mice boosted with SopB5-GVNPs, indicating that the C-terminal portions of SopB displayed on GVNPs elicit a protective response to Salmonella infection in mice. SopB antigen-GVNPs were found to be stable at elevated temperatures for extended periods without refrigeration in Halobacterium cells. The results all together show that bioengineered GVNPs are likely to represent a valuable platform for the development of improved vaccines against Salmonella diseases.