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Research Article: Soluble multi?epitope protein vaccination leverages antigen availability to drive CD8 + T cell immunity and tumor control

Date Published: 2026-06-10

Abstract:
The shift toward precision medicine in cancer immunotherapy has increased demand for rapid, scalable production platforms for personalized vaccine antigens targeting neoantigens and tumor-associated antigens. Escherichia coli-based recombinant protein production represents a globally established system offering speed and cost-effectiveness, yet the immunogenic potential of soluble antigens produced via this platform remains incompletely characterized. Here, we systematically evaluated the capacity of E. coli-derived soluble antigen formulations to elicit anti-tumor T cell responses. Using ClearColi BL21 (DE3), a lipopolysaccharide (LPS)-truncated strain free of endotoxic contamination, we produced soluble formulations containing murine OVA-derived epitopes. In vitro antigen presentation assays revealed minimal T cell activation despite high numbers of CD8 + and CD4 + T cells, indicating compromised antigen presentation capacity. However, in vivo adoptive transfer experiments demonstrated strongly inducible T cell expansion in draining and non-draining lymph nodes, with high frequencies of antigen-specific CD8 + T cells producing IFN-? and TNF-?. Prime-boost vaccination strategies in experimental melanoma models achieved protective efficacy and durable survival with persistent antigen-specific T cell responses throughout the observation period. Under stringent single-prime conditions, transient T cell frequencies were observed without long-term tumor control.

Introduction:
T cells play a central role in defending the body against intracellular infections, dysfunctional host cells, and malignant transformations such as cancer. This immune surveillance relies on the detection of antigens presented by major histocompatibility complex class I (MHC-I) molecules. While natural infections typically trigger these responses, vaccines can be designed to mimic such signals and activate antigen-specific T cells in the absence of real pathogens. In cancer immunotherapy, tumor-associated…

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