Immunological Characterization of a Nanoliposome-Formulated Peptide Vaccine Designed to Target CEA and TAG-72 Epitopes in HT29 Colorectal Cancer Cells
Archives of Advances in Biosciences,
Vol. 17 No. 1 (2026),
28 April 2026
,
Page 1-23
https://doi.org/10.22037/aab.v17i1.52477
Abstract
Background: Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide. Carcinoembryonic antigen (CEA) and tumor-associated glycoprotein 72 (TAG-72) are two well-established biomarkers commonly overexpressed in CRC. In this study, we designed a novel peptide vaccine targeting immunodominant B-cell epitopes derived from both CEA and TAG-72, formulated using a nanoliposomal delivery system, and evaluated its immunological and antitumor effects in vivo.
Methods: Using immunoinformatics tools, we predicted B-cell epitopes from the full-length sequences of CEA and TAG-72. Candidate peptides were selected based on antigenicity, immunogenicity, MHC class I and II binding affinity, and safety profiles (non-allergenic and non-toxic). Molecular dynamics simulations and docking analyses were performed to assess structural stability and TLR4-binding affinity. The optimal peptide was synthesized, encapsulated in nanoliposomes, and characterized for size, morphology, and encapsulation efficiency. Female BALB/c mice were immunized with different doses of the nanolipopeptide formulation, and immune responses were evaluated by measuring antibody titers, cytokine levels, antibody binding to HT29 cells, apoptosis induction, and expression of apoptosis-related genes (BAX and BCL2).
Results: he multi-epitope peptides, one from CEA (YVCGIQNSVS) and one from TAG-72 (TAPVTSGPVS), exhibited strong antigenicity, favorable immunogenicity, and broad MHC-binding potential. Molecular simulations supported their structural stability and efficient binding to TLR4. The nanolipopeptide particles demonstrated a uniform size (~125 nm), spherical morphology, and satisfactory encapsulation efficiency (76%). Immunization with the highest dose elicited robust antibody responses, with endpoint titers exceeding 1:10,000. Significant increases in IFN-γ levels and marked apoptosis of HT29 cells (approximately 39%) were observed in vaccinated groups compared with controls. Additionally, BAX expression was significantly upregulated, while BCL2 expression was downregulated, indicating activation of the intrinsic apoptotic pathway.
Conclusion: Our findings demonstrate that a nanoliposome-formulated peptide vaccine effectively elicits strong humoral and cellular immune responses, promotes antibody-mediated recognition of CRC cells, and induces apoptosis through the intrinsic pathway.
- Colorectal neoplasms, peptide-based immunotherapy, nanoliposomal delivery, carcinoembryonic antigen, TAG-72 glycoprotein, programmed cell death, HT29 cells
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References
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