Determination of cell expansion and surface molecule expression on anti‐CD3/28 expanded CD4⁺ T cells

Abstract

CD4+ T cell immunotherapy has potential for treatment in HIV‐infected patients. A large number of expanded CD4+ T cells and confirmation of functional‐related phenotypes are required for ensuring the successful outcomes of treatment. Freshly isolated CD4+ T cells from healthy donors were activated with anti‐CD3/28‐coated magnetic beads at different bead‐to‐cell ratios and cultured in the absence and presence of IL‐2 supplementation for 3 weeks. Fold expansion, cell viability, growth kinetic and lymphocyte subset identities were determined. Data demonstrated that a 1:1 bead‐to‐cell ratio rendered the highest expansion of 1044‐fold with 88% viability and 99.5% purity followed by the 2:1 and 0.5:1 ratios. No significant difference in proliferation and phenotypes was found between non–IL‐2 and IL‐2 supplementation groups. Several specific surface molecule expressions of the expanded cells including chemokine receptors, adhesion molecules, co‐stimulatory molecules, activation molecules, maturation markers, cytokine receptors and other molecules were altered when compared to the unexpanded cells. This optimized expansion protocol using the 1:1 bead‐to‐cell ratio of anti‐CD3/28‐coated magnetic beads and culture condition without IL‐2 supplementation provided the satisfactory yield with good reproducibility. Specific surface molecule expressions of the expanded cells presented potential roles in proliferation, differentiation, homeostasis, apoptosis and organ homing.

Nanomaterials Boost CAR‐T Therapy for Solid Tumors

“…[80] Within the scope of T cell expansion, magnetic particle aAPCs conjugated with anti-CD28 (𝛼CD28) and anti-CD3 antibodies (𝛼CD3) have emerged as fundamental to both clinical and scientific research. [81][82][83] However, this method has limitations, such as inducing an effector state in T cells before transfusion, resulting in decreased viability. [84,85] To overcome these limitations, Roh's team enhanced T cell viability by incorporating a molecular modification of HER2 onto the surface of iron oxide particles (Dynabeads), creating a cell-free artificial target particle (aaTP) that induces CAR-T cell activation (Figure 3a).…”