Gilfillan Darby, Vilander Allison C, Pan Meichen, Goh Yong Jun, O'Flaherty Sarah, Feng Ningguo, Fox Bridget E, Lang Callie, Greenberg Harry B, Abdo Zaid, Barrangou Rodolphe, Dean Gregg A
Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC 27695, USA.
Vaccines (Basel). 2023 Nov 28;11(12):1774. doi: 10.3390/vaccines11121774.
Rotavirus diarrhea-associated illness remains a major cause of global death in children under five, attributable in part to discrepancies in vaccine performance between high- and low-middle-income countries. Next-generation probiotic vaccines could help bridge this efficacy gap. We developed a novel recombinant (rLA) vaccine expressing rotavirus antigens of the VP8* domain from the rotavirus EDIM VP4 capsid protein along with the adjuvants FimH and FliC. The -based counterselective gene-replacement system was used to chromosomally integrate FimH, VP8Pep (10 amino acid epitope), and VP8-1 (206 amino acid protein) into the genome, with FliC expressed from a plasmid. VP8 antigen and adjuvant expression were confirmed by flow cytometry and Western blot. Rotavirus naïve adult BALB/cJ mice were orally immunized followed by murine rotavirus strain EC viral challenge. Antirotavirus serum IgG and antigen-specific antibody-secreting cell responses were detected in rLA-vaccinated mice. A day after the oral rotavirus challenge, fecal antigen shedding was significantly decreased in the rLA group. These results indicate that novel rLA constructs expressing VP8 can be successfully constructed and used to generate modest homotypic protection from rotavirus challenge in an adult murine model, indicating the potential for a probiotic next-generation vaccine construct against human rotavirus.
轮状病毒腹泻相关疾病仍然是全球五岁以下儿童死亡的主要原因，部分原因是高收入和中低收入国家疫苗效果存在差异。新一代益生菌疫苗可能有助于弥合这一疗效差距。我们开发了一种新型重组（rLA）疫苗，该疫苗表达来自轮状病毒EDIM VP4衣壳蛋白VP8*结构域的轮状病毒抗原以及佐剂FimH和FliC。基于的反选择基因替换系统用于将FimH、VP8Pep（10个氨基酸表位）和VP8-1（206个氨基酸蛋白）染色体整合到基因组中，FliC从质粒表达。通过流式细胞术和蛋白质印迹法确认了VP8抗原和佐剂的表达。对未接触过轮状病毒的成年BALB/cJ小鼠进行口服免疫，随后用鼠轮状病毒株EC进行病毒攻击。在接种rLA疫苗的小鼠中检测到抗轮状病毒血清IgG和抗原特异性抗体分泌细胞反应。口服轮状病毒攻击一天后，rLA组的粪便抗原排出量显著降低。这些结果表明，表达VP8的新型rLA构建体可以成功构建，并用于在成年小鼠模型中产生适度的同型轮状病毒攻击保护，这表明了一种针对人类轮状病毒的益生菌下一代疫苗构建体的潜力。
