Development and Experimental Evaluation of a Multi-Epitope Nanolipopeptide Platform Targeting PD-1, PD-L1, and HER2 for Gastric Adenocarcinoma Immunotherapy
Archives of Advances in Biosciences,
Vol. 17 No. 1 (2026),
28 April 2026
,
Page 1-15
https://doi.org/10.22037/aab.v17i1.52476
Abstract
Background and Aim: To design and experimentally validate a multi-epitope nanolipopeptide vaccine targeting three key molecules—PD-1, PD-L1, and HER2—for immunotherapy of gastric adenocarcinoma.
Methods: Using immunoinformatics, linear B cell epitopes were predicted from each target protein and screened for antigenicity, allergenicity, toxicity, and MHC binding. The selected epitopes were concatenated into a single construct, docked against TLR4, and subjected to 30 ns molecular dynamics simulation. The final peptide was encapsulated in nanoliposomes and administered subcutaneously to BALB/c mice (three doses: 25, 50, 100 mg/mL). Immune responses were measured by ELISA and cytometric bead array. MKN-45 gastric cancer cells were used to assess antibody recognition (immunocytochemistry), apoptosis (Annexin V/PI flow cytometry), and BAX/BCL2 expression (qRT PCR).
Results: The nanolipopeptide particles averaged 120 nm in diameter (PDI = 0.23, zeta potential −28.5 mV, encapsulation efficiency 78.4%). Vaccination produced dose‑dependent anti‑peptide IgG titers up to 1:12,800. Cytokine analysis revealed elevated IFN‑γ (320 pg/mL), TNF‑α (195 pg/mL), IL‑6 (180 pg/mL), and moderate increases in IL‑4 and IL‑10. Sera from immunized animals recognized MKN-45 cell surfaces and induced 42.3% total apoptosis in the high‑dose group. Mechanistically, BAX expression rose 4.2‑fold while BCL2 dropped 3.8‑fold, indicating intrinsic pathway engagement.
Conclusion: This computer‑designed, nanoliposome‑delivered multi‑epitope vaccine elicited robust humoral and cellular immunity, enabled tumor cell recognition, and activated mitochondrial apoptosis. The platform warrants further translational investigation.
- Multi Epitope
- Nanolipopeptide
- PD 1
- PD L1
- HER2
- Immunotherapy
How to Cite
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