Haloarchaeal gas vesicle nanoparticles displaying Salmonella antigens as a novel approach to vaccine development.
Haloarchaeal gas vesicle nanoparticles displaying Salmonella antigens as a novel approach to vaccine development.
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DOI：
10.1016/j.provac.2015.05.003
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
DasSarma S
中科院分区：
文献类型：
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作者：
DasSarma P;Negi VD;Balakrishnan A;Kim JM;Karan R;Chakravortty D;DasSarma S
迫切需要一种安全、有效和廉价的疫苗来对抗伤寒和其他沙门氏菌疾病。为了解决这一需求，我们正在开发一种新的疫苗平台，该平台采用来自嗜盐古菌嗜盐杆菌NRC-1的漂浮的自辅助性气泡纳米颗粒（GVNPs），其经生物工程改造以展示高度保守的肠道沙门氏菌抗原。作为用于测试的初始抗原，我们选择SopB，一种由致病性S.肠道细菌在感染宿主细胞期间。将两个高度保守的sopB基因片段（sopB 4和sopB 5）分别与gvpC基因融合，通过离心加速浮选法纯化得到的SopB-GVNPs。使用针对SopB的短合成肽产生的抗血清，通过Western印迹分析建立SopB 4和SopB 5抗原表位在GVNPs上的展示。通过将SopB-GVNPs腹腔注射给已用S.肠道血清型鼠伤寒沙门氏菌14028 ΔpmrG-HM-D（DV-STM-07），一种减毒活疫苗株。促炎细胞因子IFN-γ、IL-2和IL-9在用SopB 5-GVNP加强的小鼠中被显著诱导，与稳健的Th 1应答一致。用S.肠血清型鼠伤寒沙门氏菌14028中，发现用SopB 4-GVNP加强的小鼠的脾脏中的细菌负荷减少，而用SopB 5-GVNP加强的小鼠的肝脏、肠系膜淋巴结和脾脏中的细菌负荷不存在或显著减少，表明展示在GVNP上的SopB的C末端部分在小鼠中引起对沙门氏菌感染的保护性应答。还发现SopB抗原-GVNPs在高温下在不冷藏的情况下长时间稳定。结果表明，生物工程GVNPs可能代表一个有价值的平台，用于抗原递送和开发针对沙门氏菌和其他疾病的改进疫苗。
A safe, effective, and inexpensive vaccine against typhoid and other Salmonella diseases is urgently needed. In order to address this need, we are developing a novel vaccine platform employing buoyant, self-adjuvanting gas vesicle nanoparticles (GVNPs) from the halophilic archaeon Halobacterium sp. NRC-1, bioengineered to display highly conserved Salmonella enterica antigens. As the initial antigen for testing, we selected SopB, a secreted inosine phosphate effector protein injected by pathogenic S. enterica bacteria during infection into the host cells. Two highly conserved sopB gene segments near the 3′-region, named sopB4 and sopB5, were each fused to the gvpC gene, and resulting SopB-GVNPs were purified by centrifugally accelerated flotation. Display of SopB4 and SopB5 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 SopB-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 also found to be stable at elevated temperatures for extended periods without refrigeration. The results show that bioengineered GVNPs are likely to represent a valuable platform for antigen delivery and development of improved vaccines against Salmonella and other diseases.