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Cell-mediated Immune Responses01:40

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T cells are integral to our adaptive immune system, recognizing and effectively responding to foreign antigens. T cell activation and clonal selection are pivotal in orchestrating this immune response. This article elucidates these mechanisms, detailing the roles of cluster of differentiation (CD) markers, major histocompatibility complex (MHC) molecules, costimulatory signals, and the process of clonal selection.
Naive T cells that have not yet encountered an antigen express two primary CD...

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Hilde Almåsbak1, Edith Rian, Hanna Julie Hoel

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Messenger RNA (mRNA) transfection offers a transient method for engineering T cells with chimeric antigen receptors (CARs) for cancer therapy. This approach enables rapid pre-clinical screening and enhances safety for early-phase clinical trials.

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Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • T cell redirection using chimeric antigen receptors (CARs) or bispecific T cell engagers (BiTEs) is a strategy to combat cancer.
  • Messenger RNA (mRNA) transfection presents a novel alternative for transient T cell modification.

Purpose of the Study:

  • To evaluate the pre-clinical efficacy of transient T cell modification with CD19 CAR mRNA for B-cell malignancies.
  • To assess the safety and efficiency of mRNA electroporation for CAR delivery into T cells.

Main Methods:

  • Generated CAR mRNA using a scalable, cell-free process with recombinant RNA polymerase.
  • Employed non-toxic square-wave electroporation for efficient mRNA loading into T cells, avoiding insertional mutagenesis.
  • Assessed T cell viability, CAR expression, and cytolytic activity against CD19+ targets.

Main Results:

  • Achieved >80% T cell viability and 94% CAR expression post-transfection.
  • Demonstrated T cell-mediated killing of CD19+ targets and interferon-gamma (IFN-γ) production.
  • Showcased successful co-electroporation of additional transgenes (CXCR4, CCR7) to modify chemotaxis.

Conclusions:

  • Transient CAR mRNA transfection is a safe and effective strategy for early-phase T cell therapy studies.
  • This methodology facilitates rapid pre-clinical evaluation of novel CAR targets and receptors.
  • Transient modification offers a safer alternative to stable transduction, especially before CAR safety is established.