Loading of human car t-cells with superparamagnetic iron-based particles for magnetic targeting

The loaded CAR T-cell for use of any of the preceding claims, wherein the loaded CAR T-cell is a 2nd generation CAR T-cell, preferably wherein said 2nd generation CAR T-cell comprises a co-stimulatory domain and an intracellular signaling domain, wherein the co-stimulatory domain comprises sections of CD28 or 4-1BB, more preferably wherein the co-stimulatory domain comprises section of CD28, and even more preferably wherein the co-stimulatory domain comprises section of CD28 and the intracellular signaling domain comprises section of CD37, thereby forming a CD28-CD3Z CAR.

The loaded CAR T-cell for use of any of the preceding claims, wherein the loaded CAR T-cell comprises from 0.5 to 4 pg iron per cell (Fe/cell), preferably from 0.55 to 3.5 pg Fe/cell, more preferably from 0.6 to 3.3 pg Fe/cell, more preferably from 0.65 to 3.1 pg Fe/cell, more preferably from 0.7 to 2.9 pg Fe/cell, more preferably from 0.8 to 2.7 pg Fe/cell, more preferably from 0.9 to 2.6 pg Fe/cell, more preferably from 1 to 2.5 pg Fe/cell, and most preferably around 1 pg Fe/cell.

The loaded CAR T-cell for use of any of the preceding claims, wherein the loaded CAR T-cell is a CSPG4-specific, a HER2-specific, a EGFRVIII-specific, a PSMA-specific, a CEA-specific, a MUC1-specific, a CD19-specific, a CD20-specific, a CD22-specific, a CD30-specific, a CD269-specific, a CD138-specific CAR T-cell, preferably wherein the loaded CAR T-cell is the CSPG4-specific CAR T-cell.

The loaded CAR T-cell for use of any of the preceding claims, wherein the loaded CAR T-cell is a theranostic agent, preferably wherein the theranostic agent is visible by MRI and/or ultrasound, more preferably wherein the MRI and/or ultrasound allows imaging controlled therapy.

The loaded CAR T-cell for use of any of the preceding claims, wherein the superparamagnetic iron-based particles have been cleaned of excess molecules by aqueous washing steps based on porous membrane filtration.

The loaded CAR T-cell for use of any of the preceding claims, wherein the superparamagnetic iron-based particles are selected from an iron oxide, iron (Fe), iron-cobalt, alnico, permalloy particles, preferably wherein the superparamagnetic iron-based particles are superparamagnetic iron oxide nanoparticles (SPIONs).

The loaded CAR T-cell for use of any of the preceding claims, wherein the superparamagnetic iron-based particles have been coated with citrate, oxalic acid, ascorbic acid, fatty acids and/or amino acids, preferably citrate.

The loaded CAR T-cell for use of any of the preceding claims, wherein the cell of the tumor is a cancer cell, preferably wherein the cancer cell is a solid tumor cell, more preferably wherein the solid tumor cell is a solid malignant tumor cell.

A CAR T-cell containing superparamagnetic iron-based particles (a ‘loaded CAR T-cell’), wherein said loaded CAR T-cell exhibits a reduced cytokine release upon binding to a cell of the tumor expressing an antigen being recognized by the CAR of the CAR T-cell, compared to a CAR T-cell not containing superparamagnetic iron-based particles (‘unloaded CAR T-cell’) under the same conditions.