Repeated PD-1/PD-L1 monoclonal antibody administration induces fatal xenogeneic hypersensitivity reactions in a murine model of breast cancer

Abstract

Monoclonal antibodies (mAbs) targeting coinhibitory molecules such as PD-1, PD-L1 and CTLA-4 are increasingly used as targets of therapeutic intervention against cancer. While these targets have led to a critical paradigm shift in treatments for cancer, these approaches are also plagued with limitations owing to cancer immune evasion mechanisms and adverse toxicities associated with continuous treatment. It has been difficult to reproduce and develop interventions to these limitations preclinically due to poor reagent efficacy and reagent xenogenecity not seen in human trials. In this study, we investigated adverse effects of repeated administration of PD-1 and PD-L1 mAbs in the murine 4T1 mammary carcinoma model. We observed rapid and fatal hypersensitivity reactions in tumor bearing mice within 30-60 min after 4-5 administrations of PD-L1 or PD-1 mAb but not CTLA-4 antibody treatment. These events occurred only in mice bearing the highly inflammatory 4T1 tumor and did not occur in mice bearing noninflammatory tumors. We observed that mortality was associated with systemic accumulation of IgG1 antibodies, antibodies specific to the PD-1 mAb, and accumulation of Gr-1 high neutrophils in lungs which have been implicated in the IgG mediated pathway of anaphylaxis. Anti-PD-1 associated toxicities were alleviated when PD-1 blockade was combined with the therapeutic HSP90 inhibitor, ganetespib, which impaired immune responses toward the xenogeneic PD-1 mAb. This study highlights a previously uncharacterized fatal hypersensitivity exacerbated by the PD-1/PD-L1 axis in the broadly used 4T1 tumor model as well as an interesting relationship between this particular class of checkpoint blockade and tumor-dependent immunomodulation.

Combination with SGT-53 overcomes tumor resistance to a checkpoint inhibitor

“…Others have also observed this fatal hypersensitivity after repeated injections of anti-PD1 (clone J43; hamster IgG) or anti-PD-L1 (clone 10F.9G2; rat IgG) in the syngeneic 4T1 tumor model and suggested that mortality was associated with neutrophil-mediated anaphylaxis involving accumulation of neutrophils in the lungs. 22 Surprisingly, we observed that this fatal hypersensitivity seen with anti-PD1 antibody was not seen when the PD1 blockade was in combination with SGT-53 (Figure 8A). Autopsies of our mice that had received five injections with anti-PD1 revealed massive infiltration of neutrophils and macrophages in the lung and liver (Figure 8B), and FACS analysis confirmed abnormalities reflecting neutrophil and macrophage infiltration in the lung (Figure 8C).…”

“…Hypersensitivity leading to death after repeated injections of xenogeneic anti-PD1 (clone J43; hamster IgG) or anti-PD-L1 (clone 10F.9G2; rat IgG) has also been observed by others using the 4T1 tumor model in BALB/c mice. 22 Increased accumulation of neutrophils in the lung and the neutrophil-induced anaphylaxis were identified as the cause of death. 22 We saw an abnormal lung infiltration of neutrophils only with anti-PD1 antibody monotherapy but not with either SGT-53 alone or with the combination of anti-PD1 and SGT-53 treatment.…”

“…22 Increased accumulation of neutrophils in the lung and the neutrophil-induced anaphylaxis were identified as the cause of death. 22 We saw an abnormal lung infiltration of neutrophils only with anti-PD1 antibody monotherapy but not with either SGT-53 alone or with the combination of anti-PD1 and SGT-53 treatment. Likewise, mortality of mice seen after treatment with anti-PD1 antibody alone was prevented when SGT-53 was given in conjunction with the checkpoint inhibitor.…”

Development of shellfish allergy after exposure to dual immune checkpoint blockade

“…IL-2 has been reported to promote hypersensitivity reactions in patients receiving iodine containing radiographic contrast media at a higher incident than the general population [12,13]. Preclinical studies highlighted fatal hypersensitivity reactions in mice with 4T1 tumors exacerbated by blockage of PD-1/PD-L1 [14]. Adverse reactions to shellfish can be produced by immunological and nonimmunological reactions.…”

Murinization and H Chain Isotype Matching of the Anti-GITR Antibody DTA-1 Reduces Immunogenicity-Mediated Anaphylaxis in C57BL/6 Mice

“…A similar hypersensitivity/ADA reaction has been recently described after a repetitive i.p. dosing with murine Ab treatments against checkpoint inhibitors (hamster anti-PD-1 and rat IgG2b anti-PD-L1) in a 4T1 tumor syngeneic model because of the xenogeneic nature of the Ab treatments (55). Strategies to mitigate risk factors from the clinical use of human protein therapeutics (i.e., humanization, CDR modification, formulation, biophysical characteristics, routes of delivery, etc.)…”