Tumor Antigen-Tethered Spiked Virus-Like- Poly(Lactic-Co-Glycolic Acid)-Nanoparticle Vaccine Enhances Antitumor Ability Through Th9 Promotion in Mice

Ting-Wei Lin,1 Po-Yu Chou,2 Yen-Ting Shen,2 Ming-Thau Sheu,2 Kuo-Hsiang Chuang,3,4 Shyr-Yi Lin,5,6 Chia-Yi Chang1 1Department of Veterinary Medicine, National Taiwan University, Taipei, Taiwan; 2School of Pharmacy, College of Pharmacy, Taipei Medical University, Taipei, Taiwan; 3PhD Program in Clinical Drug Development of Chinese Herbal Medicine, Taipei Medical University, Taipei, Taiwan; 4Graduate Institute of Pharmacognosy, Taipei Medical University, Taipei, Taiwan; 5Division of Gastroenterology and Hepatology, Department of Internal Medicine, Taipei Medical University Hospital, Taipei, Taiwan; 6Department of General Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei, TaiwanCorrespondence: Shyr-Yi Lin; Chia-Yi Chang, Email [email protected]; [email protected]: Immunotherapy emerges as a promising frontier in cancer therapy and prevention. This study investigates the capacity of tumor-antigenic nanoparticles, specifically ovalbumin-tethered spiked virus-like poly(lactic-co-glycolic acid) nanoparticles (OVA-sVLNP), to effectively elicit humoral and cellular immune responses against tumors.Methods: OVA-sVLNP were synthesized through thiol-maleimide crosslinking using a single emulsion method. Comprehensive characterization was performed through Nuclear Magnetic Resonance (NMR), dynamic light scattering, Cryo-electron microscopy (Cryo-EM), confocal microscopy, and flow cytometry. Immunogenicity was evaluated using an enzyme-linked immunosorbent assay (ELISA) for quantifying immunoglobulin levels (IgG, IgG1, IgG2a) and cytokines in mouse sera. Flow cytometry profiled cellular immune responses in mouse spleens, and organ biosafety was assessed using immunohistochemistry and hematoxylin and eosin (H&E) staining.Results: OVA-sVLNP had a mean particle size of 193.8 ± 11.9 nm, polydispersity index of 0.307 ± 0.04, and zeta potential of − 39.6 ± 10.16 mV, remaining stable for one month at 4°C. In vitro studies revealed significant upregulation of CD80/CD86 in dendritic cells, indicating robust activation. In vivo, the optimal concentration (V25) induced potent IgG, IgG1, and IgG2a antibodies, significant populations of CD3+CD4+, CD3+CD8+, and a rare subset of CD3+CD4+CD8+ memory T cells. Notably, Th9 induction resulted in the secretion of IL-9, IL-10, and other cytokines, which are crucial for orchestrating cytotoxic T cell activity and antitumor effects. Overall, higher doses did not improve outcomes, highlighting the significance of optimal dosing.Conclusion: This study demonstrated potent immunogenicity of OVA-sVLNP, characterized by the induction of specific IgG antibodies and the stimulation of cellular immune responses, particularly tumor-killing Th9 cells. The simplicity and cost-effectiveness of the manufacturing process augment the potential of OVA-sVLNP as a viable candidate for antitumor vaccines, opening new avenues for cancer prevention and cell-based therapeutic strategies. Keywords: tumor antigen-spiked virus-like nanoparticles, PLGA nanoparticles, cancer vaccine, ovalbumin, T helper 9, antitumor