Perforin-deficient CAR T cells recapitulate late-onset inflammatory toxicities observed in patients.

Abstract

Late-onset inflammatory toxicities resembling hemophagocytic lymphohistiocytosis (HLH) or macrophage activation syndrome (MAS) occur after chimeric antigen receptor T cell (CAR T cell) infusion and represent a therapeutic challenge. Given the established link between perforin deficiency and primary HLH, we investigated the role of perforin in anti-CD19 CAR T cell efficacy and HLH-like toxicities in a syngeneic murine model. Perforin contributed to both CD8+ and CD4+ CAR T cell cytotoxicity but was not required for in vitro or in vivo leukemia clearance. Upon CAR-mediated in vitro activation, perforin-deficient CAR T cells produced higher amounts of proinflammatory cytokines compared with WT CAR T cells. Following in vivo clearance of leukemia, perforin-deficient CAR T cells reexpanded, resulting in splenomegaly with disruption of normal splenic architecture and the presence of hemophagocytes, which are findings reminiscent of HLH. Notably, a substantial fraction of patients who received anti-CD22 CAR T cells also experienced biphasic inflammation, with the second phase occurring after the resolution of cytokine release syndrome, resembling clinical manifestations of HLH. Elevated inflammatory cytokines such as IL-1β and IL-18 and concurrent late CAR T cell expansion characterized the HLH-like syndromes occurring in the murine model and in humans. Thus, a murine model of perforin-deficient CAR T cells recapitulated late-onset inflammatory toxicities occurring in human CAR T cell recipients, providing therapeutically relevant mechanistic insights.

• Authors: Kazusa Ishii, Marie Pouzolles, Christopher D Chien, Rebecca A Erwin-Cohen, M Eric Kohler, Haiying Qin, Haiyan Lei, Skyler Kuhn, Amanda K Ombrello, Alina Dulau-Florea, Michael A Eckhaus, Haneen Shalabi, Bonnie Yates, Daniel A Lichtenstein, Valérie S Zimmermann, Taisuke Kondo, Jack F Shern, Howard A Young, Naomi Taylor, Nirali N Shah, Terry J Fry
• Journal: The Journal of clinical investigation (J Clin Invest) Vol. 130 Issue 10 Pg. 5425-5443 (10 01 2020) ISSN: 1558-8238 [Electronic] United States
• PMID: 32925169 (Publication Type: Journal Article, Research Support, N.I.H., Intramural)

Chemical References

• Cytokines
• Inflammation Mediators
• Receptors, Chimeric Antigen
• perforin 1, mouse
• Perforin

Topics

• Animals
• Cytokines (biosynthesis)
• Disease Models, Animal
• Humans
• Immunotherapy, Adoptive (adverse effects)
• In Vitro Techniques
• Inflammation Mediators (metabolism)
• Lymphohistiocytosis, Hemophagocytic (etiology, immunology, pathology)
• Macrophage Activation Syndrome (etiology, immunology, pathology)
• Mice
• Mice, Inbred C57BL
• Mice, Knockout
• Models, Immunological
• Perforin (deficiency, genetics)
• Receptors, Chimeric Antigen (immunology)
• T-Lymphocytes (immunology, pathology)