A multivariate, quantitative assay that disentangles key kinetic parameters of primary human T cell function in vitro

Abstract

Cell therapy is poised to play a larger role in medicine, most notably for immuno-oncology. Despite the recent success of CAR-T therapeutics in the treatment of blood tumors and the rapid progress toward improved versions of both CAR- and TCR-Ts, important analytical aspects of preclinical development and manufacturing of engineered T cells remain immature. One limiting factor is the absence of robust multivariate assays to disentangle key parameters related to function of engineered effector cells, especially in the peptide-MHC (pMHC) target realm, the natural ligand for TCRs. Here we describe an imaging-based primary T cell assay that addresses several of these limitations. To our knowledge, this assay is the first quantitative, high-content assay that separates the key functional parameters of time- and antigen-dependent T cell proliferation from cytotoxicity. We show that the assay sheds light on relevant biology of CAR- and TCR-T cells, including response kinetics and the influence of effector:target ratio.

Potent, Selective CARs as Potential T-Cell Therapeutics for HPV-positive Cancers

“…6C). [60][61][62] In addition, we confirmed the E6 CAR (CT512), E6 TCR, and E7 TCR killed the HPV-positive tumor cell line CaSki similarly at multiple E:T ratios, while they had little effect on the HPV-negative cell line A375 (Fig. 6B).…”

Re-examination of MAGE-A3 as a T-cell Therapeutic Target

“…QuIK assay method was the same as described elsewhere. 32 Adherent target cells (MCF7) were cultured under standard conditions and seeded at 2000 cells per well in LymphoONE media containing 1% human serum in 384-well plates (Greiner 781091). Serially diluted peptide solutions were added to the target cells to yield peptide concentrations ranging from ∼2 pM to 100 μM.…”

Industry Updates From the Field of Stem Cell Research and Regenerative Medicine in November 2020

“…A2 Biotherapeutics (CA, USA; www.a2biotherapeutics.com) developed an imaging-based quantitative, highcontent assay that separates the key functional parameters of time-and antigen-dependent T cell proliferation from cytotoxicity [10]. The assay, first of its kind, sheds light on relevant biology of CAR-and TCR-T cells, including response kinetics and the influence of effector: target ratio.…”