Dissecting the Mechanisms Underlying the Cytokine Release Syndrome (CRS) Mediated by T-Cell Bispecific Antibodies

Clin Cancer Res. 2023 Nov 1;29(21):4449-4463. doi: 10.1158/1078-0432.CCR-22-3667.

Abstract

Purpose: Target-dependent TCB activity can result in the strong and systemic release of cytokines that may develop into cytokine release syndrome (CRS), highlighting the need to understand and prevent this complex clinical syndrome.

Experimental design: We explored the cellular and molecular players involved in TCB-mediated cytokine release by single-cell RNA-sequencing of whole blood treated with CD20-TCB together with bulk RNA-sequencing of endothelial cells exposed to TCB-induced cytokine release. We used the in vitro whole blood assay and an in vivo DLBCL model in immunocompetent humanized mice to assess the effects of dexamethasone, anti-TNFα, anti-IL6R, anti-IL1R, and inflammasome inhibition, on TCB-mediated cytokine release and antitumor activity.

Results: Activated T cells release TNFα, IFNγ, IL2, IL8, and MIP-1β, which rapidly activate monocytes, neutrophils, DCs, and NKs along with surrounding T cells to amplify the cascade further, leading to TNFα, IL8, IL6, IL1β, MCP-1, MIP-1α, MIP-1β, and IP-10 release. Endothelial cells contribute to IL6 and IL1β release and at the same time release several chemokines (MCP-1, IP-10, MIP-1α, and MIP-1β). Dexamethasone and TNFα blockade efficiently reduced CD20-TCB-mediated cytokine release whereas IL6R blockade, inflammasome inhibition, and IL1R blockade induced a less pronounced effect. Dexamethasone, IL6R blockade, IL1R blockade, and the inflammasome inhibitor did not interfere with CD20-TCB activity, in contrast to TNFα blockade, which partially inhibited antitumor activity.

Conclusions: Our work sheds new light on the cellular and molecular players involved in cytokine release driven by TCBs and provides a rationale for the prevention of CRS in patients treated with TCBs. See related commentary by Luri-Rey et al., p. 4320.

MeSH terms

  • Animals
  • Antibodies, Bispecific* / pharmacology
  • Chemokine CCL3
  • Chemokine CCL4
  • Chemokine CXCL10
  • Cytokine Release Syndrome
  • Cytokines
  • Dexamethasone / pharmacology
  • Endothelial Cells
  • Humans
  • Inflammasomes
  • Interleukin-6
  • Interleukin-8
  • Mice
  • RNA
  • T-Lymphocytes
  • Tumor Necrosis Factor-alpha*

Substances

  • Chemokine CCL3
  • Tumor Necrosis Factor-alpha
  • Chemokine CCL4
  • Antibodies, Bispecific
  • Interleukin-8
  • Chemokine CXCL10
  • Interleukin-6
  • Inflammasomes
  • Cytokines
  • Dexamethasone
  • RNA