Single-cell technologies for profiling T cells to enable monitoring of immunotherapies

Abstract

Immunotherapy relies on the reinvigoration of immune system to combat diseases and has transformed the landscape of cancer treatments. Clinical trials using immune checkpoint inhibitors (ICI), and adoptive transfer of genetically modified T cells have demonstrated durable remissions in subsets of cancer patients. A comprehensive understanding of the polyfunctionality of T lymphocytes in ICI or adoptive cell transfer (ACT), at single-cell resolution, will quantify T-cell properties that are essential for therapeutic benefit. We briefly highlight several emerging integrated single-cell technologies focusing on the profiling of multiple properties/functionalities of T cells. We envision that these tools have the potential to provide valuable experimental and clinical insights on T-cell biology, and eventually pave the road for the discovery of surrogate T-cell biomarkers for immunotherapy.

A Microfluidic Platform for Single Cell Fluorometric Granzyme B Profiling

“…The elevated GrB production that was observed in peripheral blood mononuclear cells isolated from patients could provide information about the disease and thus shows the platform's potential utility for diagnosis. Although it is assumed that the elevated GrB levels are due to increased expression by CTL activation; the contribution from other immune cells is lacking in reports 31.…”

“…By comparing the protease expression of the immune cells, specific T cell subset responsible for the anti-tumor response can be identified. These cells with overexpression of GrB has been an interest especially in immune checkpoint inhibitors immunotherapy and adaptive cell transfer therapy 31-32.…”

A design and optimization of a high throughput valve based microfluidic device for single cell compartmentalization and analysis

“…Many research works have demonstrated its advantages in having fast isolation speed and high efficiency of data acquisition. These platforms are capable of spatial control of liquid composition, cell and cell environment manipulation, and single cell handling and quantification of secreted molecules 3 – 5 . Thus, microfluidic platforms are an important tool for single cell studies as it allows the identification of rare subsets and resolution between cells with same expression patterns.…”

“…Thus, microfluidic platforms are an important tool for single cell studies as it allows the identification of rare subsets and resolution between cells with same expression patterns. This has become an important tool for single cell biological research taking advantage of massive parallel processing, streamlining of complex protocols, and enabling high-throughput biological experiments 3 , 5 .…”

Design and Optimizations of a High-throughput Valve-based Microfluidic Device for Single Cell Compartmentalization and Analysis

“…Many research works have demonstrated its advantages in having fast isolation speed and high efficiency of data acquisition. It is capable of integrating multiple workflows that enables parallelization and streamlining of complex protocols, which allows automation and control of analytical functions, cell and cell environment manipulation, and simultaneous detection and quantification of secreted molecules 3,4 . For instance, conventional approaches don't take into account the phenotypic and functional state of heterogenous cells which microfluidics technology is able to address.…”