soluble and immobilized anti-cd3/28 distinctively expand and differentiate primary human t cells: an implication for adoptive t cell therapy

Cell-based cancer therapies have led to a paradigm shift in the treatment of patients with various cancers. to date, a vast majority of cancer immunotherapies have used genetically engineered t cells to target tumors. stimulation and ex vivo expansion of t cells, as one of the crucial starting materials for t cell manufacturing, have always been a critical part of adoptive t-cell therapy (act). typically, anti-cd3 and anti-cd28 monoclonal antibodies (mabs) along with interleukin-2 (il-2), through transducing signals one, two, and three, respectively, are essential for in vitro t cell activation. terminal differentiation and replicative senescence are the main barriers of the acts during the manufacturing of engineered t cells ex vivo. in this study, we aimed to compare the t cell activation protocol that we developed in our lab (soluble anti-cd3/28 mabs) with a common t cell activation protocol (immobilized anti-cd3/soluble anti-cd28) in terms of t cell expansion, activation, immunophenotype, and cellular fate. we observed that t cells were equally expanded in both protocols. notably, our modified protocol promoted the outgrowth of cd8+ t cells postactivation. concerning the low concentrations of both soluble anti-cd3 and anti-cd28, the modified protocol could significantly enrich memory t cell subsets. in conclusion, our data demonstrated that the soluble cd3/28 mabs protocol is cost-effective and more efficient for generating more potent t cells, thereby expecting a better therapeutic outcome.