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首页> 外文期刊>Infection and immunity >Analysis of Type II Secretion of Recombinant Pneumococcal PspA and PspC in a Salmonella enterica Serovar Typhimurium Vaccine with Regulated Delayed Antigen Synthesis
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Analysis of Type II Secretion of Recombinant Pneumococcal PspA and PspC in a Salmonella enterica Serovar Typhimurium Vaccine with Regulated Delayed Antigen Synthesis

机译：调控的延迟抗原合成分析沙门氏菌血清型鼠伤寒沙门氏菌疫苗中重组肺炎球菌PspA和PspC的II型分泌

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Recombinant attenuated Salmonella vaccines (RASVs) have been used extensively to express and deliver heterologous antigens to host mucosal tissues. Immune responses can be enhanced greatly when the antigen is secreted to the periplasm or extracellular compartment. The most common method for accomplishing this is by fusion of the antigen to a secretion signal sequence. Finding an optimal signal sequence is typically done empirically. To facilitate this process, we constructed a series of plasmid expression vectors, each containing a different type II signal sequence. We evaluated the utilities of these vectors by fusing two different antigens, the α-helix domains of pneumococcal surface protein A (PspA) and pneumococcal surface protein C (PspC), to the signal sequences of β-lactamase (bla SS), ompA, and phoA and the signal sequence and C-terminal peptide of β-lactamase (bla SS+CT) on Asd+ plasmids under the control of the Ptrc promoter. Strains were characterized for level of expression, subcellular antigen location, and the capacity to elicit antigen-specific immune responses and protection against challenge with Streptococcus pneumoniae in mice. The immune responses to each protein differed depending on the signal sequence used. Strains carrying the bla SS-pspA and bla SS+CT-pspC fusions yielded the largest amounts of secreted PspA and PspC, respectively, and induced the highest serum IgG titers, although all fusion proteins tested induced some level of antigen-specific IgG response. Consistent with the serum antibody responses, RASVs expressing the bla SS-pspA and bla SS+CT-pspC fusions induced the greatest protection against S. pneumoniae challenge.
机译：重组减毒的沙门氏菌疫苗（RASVs）已广泛用于表达异源抗原并将其传递至宿主粘膜组织。当抗原分泌到周质或细胞外区室时，免疫反应可以大大增强。实现此目的的最常见方法是将抗原融合到分泌信号序列上。找到最佳信号序列通常是凭经验完成的。为促进此过程，我们构建了一系列质粒表达载体，每个载体均包含不同的II型信号序列。我们通过将两种不同的抗原（肺炎球菌表面蛋白A（PspA）和肺炎球菌表面蛋白C（PspC））的两种不同抗原融合到β-内酰胺酶（ bla SS）， ompA 和 phoA 以及β-内酰胺酶（ bla SS + CT）的信号序列和C端肽在P trc 启动子控制下的Asd + 质粒。表征了菌株的表达水平，亚细胞抗原定位，引发抗原特异性免疫反应的能力以及对小鼠肺炎链球菌攻击的保护作用。对每种蛋白质的免疫反应取决于所使用的信号序列而有所不同。携带 bla SS- pspA 和 bla SS + CT- pspC 融合蛋白的菌株产生最大量的分泌PspA尽管所有测试的融合蛋白均可诱导一定水平的抗原特异性IgG应答，但分别以PspC和PspC诱导的血清IgG滴度最高。与血清抗体反应一致的是，表达 bla SS- pspA 和 bla SS + CT- pspC 融合蛋白的RASV诱导出对 S的最大保护。肺炎挑战。

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