Submicron immunoglobulin particles exhibit FcγRII-dependent toxicity linked to autophagy in TNFα-stimulated endothelial cells

Wanida C Hollis; Sehrish Farooq; M Reza Khoshi; Mehulkumar Patel; Elena Karnaukhova; Nancy Eller; Karel Holada; Dorothy E Scott; Jan Simak

doi:10.1007/s00018-024-05342-9

Submicron immunoglobulin particles exhibit FcγRII-dependent toxicity linked to autophagy in TNFα-stimulated endothelial cells

Cell Mol Life Sci. 2024 Aug 30;81(1):376. doi: 10.1007/s00018-024-05342-9.

Authors

Affiliations

¹ Center for Biologics Evaluation and Research, FDA, Silver Spring, MD, USA.
² Center for Devices and Radiological Health, FDA, Silver Spring, MD, USA.
³ Institute of Immunology and Microbiology, First Faculty of Medicine, Charles University, Prague, Czech Republic.
⁴ Center for Biologics Evaluation and Research, FDA, Silver Spring, MD, USA. jan.simak@fda.hhs.gov.
⁵ Laboratory of Cellular Hematology, Division of Blood Components and Devices, Center for Biologics Evaluation and Research, Food and Drug Administration, OBRR, 10903 New Hampshire Avenue, WO Bldg. 52/72, Rm. 4210, Silver Spring, MD, USA. jan.simak@fda.hhs.gov.

Abstract

In intravenous immunoglobulins (IVIG), and some other immunoglobulin products, protein particles have been implicated in adverse events. Role and mechanisms of immunoglobulin particles in vascular adverse effects of blood components and manufactured biologics have not been elucidated. We have developed a model of spherical silica microparticles (SiMPs) of distinct sizes 200-2000 nm coated with different IVIG- or albumin (HSA)-coronas and investigated their effects on cultured human umbilical vein endothelial cells (HUVEC). IVIG products (1-20 mg/mL), bare SiMPs or SiMPs with IVIG-corona, did not display significant toxicity to unstimulated HUVEC. In contrast, in TNFα-stimulated HUVEC, IVIG-SiMPs induced decrease of HUVEC viability compared to HSA-SiMPs, while no toxicity of soluble IVIG was observed. 200 nm IVIG-SiMPs after 24 h treatment further increased ICAM1 (intercellular adhesion molecule 1) and tissue factor surface expression, apoptosis, mammalian target of rapamacin (mTOR)-dependent activation of autophagy, and release of extracellular vesicles, positive for mitophagy markers. Toxic effects of IVIG-SiMPs were most prominent for 200 nm SiMPs and decreased with larger SiMP size. Using blocking antibodies, toxicity of IVIG-SiMPs was found dependent on FcγRII receptor expression on HUVEC, which increased after TNFα-stimulation. Similar results were observed with different IVIG products and research grade IgG preparations. In conclusion, submicron particles with immunoglobulin corona induced size-dependent toxicity in TNFα-stimulated HUVEC via FcγRII receptors, associated with apoptosis and mTOR-dependent activation of autophagy. Testing of IVIG toxicity in endothelial cells prestimulated with proinflammatory cytokines is relevant to clinical conditions. Our results warrant further studies on endothelial toxicity of sub-visible immunoglobulin particles.

Keywords: Biologics; Contaminants; FcγR; Immunogenicity; Membrane microparticles; Protein aggregates; Protein corona; Vascular toxicity; Vessel wall.

MeSH terms

Apoptosis / drug effects
Autophagy* / drug effects
Cell Survival / drug effects
Human Umbilical Vein Endothelial Cells* / drug effects
Human Umbilical Vein Endothelial Cells* / metabolism
Humans
Immunoglobulins, Intravenous*
Intercellular Adhesion Molecule-1 / metabolism
Particle Size
Protein Corona / metabolism
Receptors, IgG* / metabolism
Silicon Dioxide / chemistry
Silicon Dioxide / toxicity
TOR Serine-Threonine Kinases / metabolism
Tumor Necrosis Factor-alpha* / metabolism

Substances

Tumor Necrosis Factor-alpha
Receptors, IgG
Immunoglobulins, Intravenous
Silicon Dioxide
Intercellular Adhesion Molecule-1
Protein Corona
TOR Serine-Threonine Kinases

Abstract

MeSH terms

Substances

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