CEA-Functionalized Gold Nanoparticles for Oral Immunoprophylaxis: In Vivo Evaluation of Safety, Biodistribution, and Immunogenicity in a Murine Colorectal Cancer Model

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Abstract

Background and aim: Colorectal cancer remains a leading cause of cancer-related mortality, with growing interest in nanotechnology-driven immunotherapeutics. Gold nanoparticles (AuNPs) offer a promising platform due to their biocompatibility, functional versatility, and immunomodulatory potential. Carcinoembryonic antigen (CEA), highly expressed in colorectal tumors, provides an ideal target for antigen-specific immune activation. The aim of this study was to evaluate the immunogenicity, biodistribution and therapeutic efficacy of a CEA-functionalised gold nanoparticle (CEA-AuNP) construct in a mouse model of colorectal cancer following oral administration via a customised capsular delivery system Methods: An in vivo study was conducted after a 30-day oral administration in BALB/c mice at escalating doses (5–50 mg/kg) of priorly validated in vitro CEA-functionalized gold nanoparticles (CEA-AuNPs). Histological, hyperspectral, and ELISA-based cytokine analyses were performed to assess tissue integrity, biodistribution, and immune responses. Results: CEA-AuNPs demonstrated high biocompatibility and effective macrophage internalization, inducing IL-10-dominant immune responses in the spleen and controlled pro-inflammatory cytokine shifts in the liver. Histological findings confirmed dose-dependent splenic and hepatic accumulation without systemic toxicity. Multifocal splenic amyloidosis and extramedullary hepatic hematopoiesis were noted at high doses, consistent with sustained immune activation. Conclusions: CEA-functionalized AuNPs represent a promising nanovaccine candidate for colorectal cancer prophylaxis. The construct induced tissue-specific immune responses, favoring cellular immunity and antigen processing without significant toxicity. These results support further investigation into CEA-AuNP nanoconstructs as oral immunotherapeutics in cancer prevention.

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