Journal
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Volume 657, Issue -, Pages 8-15Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2023.03.035
Keywords
Single cell analysis; Cell-cell-interaction; T-cell; scRNA-seq; Ca2? flux; And microfluidics
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A new non-invasive screening method has been developed to determine the activation state of T-cells at the single-cell level. By examining the intracellular Ca2+ flux and motility of T-cells interacting with Antigen Presenting Cells, this method can efficiently identify activated T-cells with good specificity and stable proliferation, which is crucial for the development of adoptive immunotherapy.
A new non-invasive screening profile has been realized that can aid in determining T-cell activation state at single-cell level. Production of activated T-cells with good specificity and stable proliferation is greatly beneficial for advancing adoptive immunotherapy as innate immunological cells are not effective in recognizing and eliminating cancer as expected. The screening method is realized by relating intracel-lular Ca2 thorn intensity and motility of T-cells interacting with APC (Antigen Presenting Cells) in a micro -fluidic chip. The system is tested using APC pulsed with OVA257-264 peptide and its modified affinities (N4, Q4, T4 and V4), and the T-cells from OT-1 mice. In addition, single cell RNA sequencing reveals the activation states of the cells and the clusters from the derived profiles can be indicative of the T-cell activation state. The presented system here can be versatile for a comprehensive application to proceed with T-cell-based immunotherapy and screen the antigen-specific T-cells with excellent efficiency and high proliferation.(c) 2023 Elsevier Inc. All rights reserved.
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