Sanchez-Martinez Zalma V, Alpuche-Lazcano Sergio P, Stuible Matthew, Akache Bassel, Renner Tyler M, Deschatelets Lise, Dudani Renu, Harrison Blair A, McCluskie Michael J, Hrapovic Sabahudin, Blouin Julie, Wang Xinyu, Schuller Matthew, Cui Kai, Cho Jae-Young, Durocher Yves
Human Health Therapeutics Research Centre, National Research Council of Canada, Montreal, QC H4P 2R2, Canada.; Department of Biochemistry and Molecular Medicine, University of Montreal, Montreal, QC H3T 1J4, Canada.
Human Health Therapeutics Research Centre, National Research Council of Canada, Montreal, QC H4P 2R2, Canada.; Current address: Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México (UNAM), A.P. 510-3, Cuernavaca Morelos (CP 62250), Mexico.
Vaccine. 2024 Dec 2;42(26):126463. doi: 10.1016/j.vaccine.2024.126463. Epub 2024 Oct 30.
A vaccine effective against both SARS-CoV-2 and influenza A (IAV) viruses could represent a cost-effective strategy to reduce their combined public health burden as well as potential complications arising from co-infection. Based on previous findings that full-length SARS-CoV-2 spike (S) expression can induce high-level, enveloped VLP (eVLP) production in CHO cells, we tested whether IAV H1N1 hemagglutinin (H1) and neuraminidase (N1) could also be displayed on these particles. We found that co-incorporation of the IAV surface antigens in spike VLPs (S-VLPs) was highly efficient: upon transient co-expression of S + H1 or S + H1 + N1 in CHO cells, the resulting VLPs contained similar amounts of the SARS-CoV-2 S and IAV antigens. The self-assembled bivalent (S/H1) and trivalent (S/H1/N1) VLPs released into the culture media were purified by single-step chromatography using a S-VLP affinity resin. Western blot analysis and immuno‑gold labeling transmission electron microscopy (TEM) of purified VLPs confirmed the coexistence of S, H1 and N1 antigens in the same particles. Finally, we demonstrated that two doses of adjuvanted bivalent and trivalent VLPs elicit specific functional antibodies and cellular immunity in a mouse model, suggesting potential for combined SARS-CoV-2/IAV vaccine development.
一种对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）和甲型流感病毒（IAV）均有效的疫苗，可能是一种具有成本效益的策略，可减轻它们共同造成的公共卫生负担以及合并感染引起的潜在并发症。基于先前的研究结果，即全长SARS-CoV-2刺突（S）蛋白的表达可在CHO细胞中诱导高水平的包膜病毒样颗粒（eVLP）产生，我们测试了IAV H1N1血凝素（H1）和神经氨酸酶（N1）是否也能展示在这些颗粒上。我们发现，IAV表面抗原在刺突病毒样颗粒（S-VLPs）中的共掺入效率很高：在CHO细胞中瞬时共表达S + H1或S + H1 + N1后，产生的病毒样颗粒含有相似量的SARS-CoV-2 S蛋白和IAV抗原。释放到培养基中的自组装二价（S/H1）和三价（S/H1/N1）病毒样颗粒，使用S-VLP亲和树脂通过单步色谱法进行纯化。对纯化的病毒样颗粒进行的蛋白质免疫印迹分析和免疫金标记透射电子显微镜（TEM）证实，S、H1和N1抗原在同一颗粒中共存。最后，我们证明，在小鼠模型中，两剂佐剂化的二价和三价病毒样颗粒可引发特异性功能性抗体和细胞免疫，这表明SARS-CoV-2/IAV联合疫苗具有开发潜力。
