Physical Characterization and Innate Immunogenicity of Aggregated Intravenous Immunoglobulin (IGIV) in an In Vitro Cell-Based Model

E. M. Moussa, J. Kotarek, Janice Blum, E. Marszal, E. M. Topp

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Purpose: To investigate in vitro the innate immune response to accelerated stress-induced aggregates of intravenous immunoglobulin (IGIV) using a well-defined human cell-line model, and to correlate the innate response to physical properties of the aggregates. Methods: IGIV aggregates were prepared by applying various accelerated stress methods, and particle size, count and structure were characterized. Immune cell activation as tracked by inflammatory cytokines released in response to aggregates was evaluated in vitro using peripheral blood mononuclear cells (PBMC), primary monocytes and immortalized human monocyte-like cell lines. Results: IGIV aggregates produced by mechanical stress induced higher cytokine release by PBMC and primary monocytes than aggregates formed by other stresses. Results with the monocytic cell line THP-1 paralleled trends in PBMC and primary monocytes. Effects were dose-dependent, enhanced by complement opsonization, and partially inhibited by blocking toll-like receptors (TLR2 and TLR4) and to a lesser extent by blocking Fc gamma receptors (FcγRs). Conclusions: Stress-induced IGIV aggregates stimulate a dose-dependent cytokine response in human monocytes and THP-1 cells, mediated in part by TLRs, FcγRs and complement opsonization. THP-1 cells resemble primary monocytes in many respects with regard to tracking the innate response to IgG aggregates. Accordingly, the measurement of inflammatory cytokines released by THP-1 cells provides a readily accessible assay system to screen for the potential innate immunogenicity of IgG aggregates. The results also highlight the role of aggregate structure in interacting with the different receptors mediating innate immunity.

Original languageEnglish (US)
Pages (from-to)1-16
Number of pages16
JournalPharmaceutical Research
DOIs
StateAccepted/In press - Apr 1 2016

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Intravenous Immunoglobulins
Monocytes
Cytokines
IgG Receptors
Blood Cells
Innate Immunity
Cell Line
Immunoglobulin G
Blood
Cells
Mechanical Stress
Toll-Like Receptors
Particle Size
In Vitro Techniques
Assays
Physical properties
Chemical activation
Particle size

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Organic Chemistry
  • Molecular Medicine
  • Pharmacology (medical)
  • Biotechnology
  • Pharmacology

Cite this

Physical Characterization and Innate Immunogenicity of Aggregated Intravenous Immunoglobulin (IGIV) in an In Vitro Cell-Based Model. / Moussa, E. M.; Kotarek, J.; Blum, Janice; Marszal, E.; Topp, E. M.

In: Pharmaceutical Research, 01.04.2016, p. 1-16.

Research output: Contribution to journalArticle

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