Abstract
The quantification of T-cell immune responses is crucial for the monitoring of natural and treatment-induced immunity, as well as for the validation of new immunotherapeutic approaches. The present study presents a simple method based on lipofection of synthetic mRNA in mononuclear cells as a method to determine in vitro T-cell responses. We compared several commercially available transfection reagents for their potential to transfect mRNA into human peripheral blood mononuclear cells and murine splenocytes. We also investigated the impact of RNA modifications in improving this method. Our results demonstrate that antigen-specific T-cell immunomonitoring can be easily and quickly performed by simple lipofection of antigen-coding mRNA in complex immune cell populations. Thus, our work discloses a convenient solution for the in vitro monitoring of natural or therapy-induced T-cell immune responses.
Highlights
Easier, and more robust method for T-cell immunomonitoring, we explored whether freshly-isolated peripheral blood mononuclear cells (PBMCs) could be transfected with mRNA using commercially available transfection reagents, including lipoplexes and polyplexes, to induce mRNA-coded epitope-specific immune responses
The transfection proficiency was determined by measuring luciferase activity 24 h post luciferase-coding ivt-mRNA lipofection, upon the addition of luciferin substrat
Synthetic mRNA has been successfully used as a vector for induction of antigenSynthetic mRNA has been successfully used as a vector for induction of antigen-spespecific immune responses in vitro and in vivo
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. The intricate complexity of the human immune system makes it challenging to analyze its responses to infection, disease, injury, or medical intervention. Immunomonitoring of T-cell responses provides information on the nature and state of immune reactions and is required to assess the efficacy of a medical treatment or to predict its effects [1]. Phenotyping of immune cell populations aids in the elucidation of the cellular mechanisms underlying newly developed immunotherapeutic approaches. It can identify the presence of cellular and molecular signatures that categorize patients into distinct risk groups and/or help to predict clinical responses to therapy [2]
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