Intranasal immunization with Ag85B peptide 25 displayed on Lactococcus lactis using the PilVax platform induces antigen-specific B- and T-cell responses,Immunology and Cell Biology

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Intranasal immunization with Ag85B peptide 25 displayed on Lactococcus lactis using the PilVax platform induces antigen-specific B- and T-cell responses
Immunology and Cell Biology ( IF 3.2 ) Pub Date : 2021-04-18 , DOI: 10.1111/imcb.12462
Samuel Blanchett ₁ , Catherine Jy Tsai _{1,

2} , Sarah Sandford ₃ , Jacelyn Ms Loh _{1,

2} , Lucy Huang ₃ , Joanna R Kirman _{2,

3} , Thomas Proft _{1,

2}

Affiliation

Mycobacterium tuberculosis (Mtb) remains a global epidemic despite the widespread use of Bacillus Calmette–Guérin (BCG). Consequently, novel vaccines are required to facilitate a reduction in Mtb morbidity and mortality. PilVax is a peptide delivery strategy for the generation of highly specific mucosal immune responses and is based on the food-grade bacterium Lactococcus lactis that is used to express selected peptides engineered within the Streptococcus pyogenes M1T1 pilus, allowing for peptide amplification, stabilization and enhanced immunogenicity. In the present study, the dominant T-cell epitope from the Mtb protein Ag85B was genetically engineered into the pilus backbone subunit and expressed on the surface of L. lactis. Western blot and flow cytometry confirmed formation of pilus containing the peptide DNA sequence. B-cell responses in intranasally vaccinated mice were analyzed by ELISA while T-cell responses were analyzed by flow cytometry. Serum titers of peptide-specific immunoglobulin (Ig) G and IgA were detected, confirming that vaccination produced antibodies against the cognate peptide. Peptide-specific IgA was also detected across several mucosal sites sampled. Peptide-specific CD4⁺ T cells were detected at levels similar to those of mice immunized with BCG. PilVax immunization resulted in an unexpected increase in the numbers of CD3⁺CD4^–CD8^– [double negative (DN)] T cells in the lungs of vaccinated mice. Analysis of cytokine production following stimulation with the cognate peptide showed the major cytokine producing cells to be CD4⁺ T cells and DN T cells. This study provides insight into the antibody and peptide-specific cellular immune responses generated by PilVax vaccination and demonstrates the suitability of this vaccine for conducting a protection study.

中文翻译：

使用 PilVax 平台在乳酸乳球菌上展示的 Ag85B 肽 25 进行鼻内免疫诱导抗原特异性 B 和 T 细胞反应

尽管卡介苗（BCG）广泛使用，结核分枝杆菌（Mtb）仍然是一种全球流行病。因此，需要新型疫苗来促进降低Mtb发病率和死亡率。PilVax 是一种用于产生高度特异性黏膜免疫反应的肽递送策略，基于食品级细菌乳酸乳球菌，用于表达在化脓性链球菌M1T1 菌毛中设计的选定肽，从而实现肽扩增、稳定和增强免疫原性. 在本研究中，来自Mtb的显性 T 细胞表位蛋白质 Ag85B 被基因工程改造到菌毛骨架亚基中，并在乳酸乳球菌的表面表达。蛋白质印迹和流式细胞术证实了含有肽 DNA 序列的菌毛的形成。通过ELISA分析鼻内接种小鼠的B细胞反应，而通过流式细胞术分析T细胞反应。检测到肽特异性免疫球蛋白 (Ig) G 和 IgA 的血清滴度，证实疫苗接种产生了针对同源肽的抗体。在取样的几个粘膜部位也检测到肽特异性 IgA。检测到的肽特异性 CD4 ⁺ T 细胞水平与用 BCG 免疫的小鼠相似。PilVax 免疫导致 CD3 ⁺ CD4数量意外增加^–CD8 ^– [双阴性 (DN)] 接种疫苗小鼠肺中的 T 细胞。用同源肽刺激后细胞因子产生的分析显示主要的细胞因子产生细胞是CD4 ⁺ T细胞和DN T细胞。该研究提供了对 PilVax 疫苗接种产生的抗体和肽特异性细胞免疫反应的深入了解，并证明了该疫苗适用于进行保护研究。

更新日期：2021-04-18

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