Protocol to measure human IL-6 secretion from CAR T cell-primed macrophage and monocyte lineage cells in vitro and in vivo using humanized mice.

Chimeric antigen receptor (CAR) T cell therapy often causes serious toxicities, such as cytokine release syndrome (CRS), mainly due to interleukin-6 (IL-6) secreted by monocyte lineage cells. Here, we describe a protocol to generate anti-CD19 CAR T cells and quantify human monocyte-derived IL-6 cocultured with CAR T cells and target tumor cells in vitro. We further describe a protocol to generate a humanized mouse model and evaluate CAR T cell-associated plasma IL-6 levels in vivo. For complete details on the use and execution of this protocol, please refer to Yoshikawa et al. ¹ .
Competing Interests: Declaration of interests This study was partly supported by a commercial research grant from Takara Bio, Inc. S.O. and Y.A. are employees of Takara Bio, Inc. T.Y. is currently an employee of Noile-Immune Biotech. Aichi Prefecture has filed a patent application related to this study on which Y.K. and T.Y. are named as inventors.
(Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)