Functional TCR T cell screening using single-cell droplet microfluidics

Abstract

Adoptive T cell transfer, in particular TCR T cell therapy, holds great promise for cancer immunotherapy with encouraging clinical results. However, finding the right TCR T cell clone is a tedious, time-consuming, and costly process. Thus, there is a critical need for single cell technologies to conduct fast and multiplexed functional analyses followed by recovery of the clone of interest. Here, we use droplet microfluidics for functional screening and real-time monitoring of single TCR T cell activation upon recognition of target tumor cells. Notably, our platform includes a tracking system for each clone as well as a sorting procedure with 100% specificity validated by downstream single cell reverse-transcription PCR and sequencing of TCR chains. Our TCR screening prototype will facilitate immunotherapeutic screening and development of T cell therapies.

Biomaterials as Tools to Decode Immunity

“…This limits the ability to detect the rare T cells that are specific for a desired antigen such as in cancer immunotherapies. To create a real‐time system to study this specificity in the context of T cell activation, Segaliny et al used soft lithography to create a PDMS microfluidic system to form droplets . Droplets were formed using a flow focusing process in which fluorocarbon oil and surfactant intersect at a right angle with an aqueous solution containing cells ( Figure a).…”

“…Briefly, T cells and cancer cells flow alongside each other before passing through oil that creates a droplet containing one cell of each type. Reproduced with permission . Copyright 2018, Royal Society of Chemistry.…”

“…This process allows for more detailed downstream analysis of cells with characteristics of interest. Reproduced with permission . Copyright 2018, Royal Society of Chemistry.…”

“…To create a real-time system to study this specificity in the context of T cell activation, Segaliny et al used soft lithography to create a PDMS microfluidic system to form droplets. [141] Droplets were formed using a flow focusing process in which fluorocarbon oil and surfactant intersect at a right angle with an aqueous solution containing cells (Figure 5a). In this system, two solutions flowed side-by-side, with one solution containing cancer cells and the other containing a heterogeneous T cell population.…”

“…Reproduced with permission. [141] Copyright 2018, Royal Society of Chemistry. b) Cells containing droplets can be sorted on the basis of cellular processes such as T cell activation.…”