A NEW PLATFORM TO DETECT INNATE IMMUNE RESPONSE MODULATING IMPURITIES (IIRMIS) IN THERAPEUTIC PROTEINS BASED ON THE USE OF MONOCYTE-DERIVED MACROPHAGES AND DENDRITIC CELLS

EDUARDO MUFARREGE; RICOTTI, SONIA; SOFIA GIORGETTI; ETCHEVERRIGARAY, MARINA; DANIELA VERTHELYI

Buenos Aires

Congreso; Reunión Conjunta 2017 entre la Sociedad Argentina de Investigación Clínica (SAIC), la Sociedad Argentina de Inmunología (SAI) y la Sociedad Argentina de Farmacología Experimental (SAFE); 2017

SAIC, SAI, SAFE

Biotherapeutics are agents produced through complex multi-step manufacturing methods. However, these products can contain contaminants derived from the host cell and manufacturing process that can trigger innate immune receptors, leading to the activation of immune cells and the secretion of cytokine and chemokines that could ultimately lead to the generation of anti-drug antibodies. To date numerous experimental approaches have been proposed to assess the risk of product immunogenicity and are mainly based on peripheral blood mononuclear cells (PBMC). However, macrophages and dendritic cells, which are rich in pattern recognition receptors (PRR) are present at very low frequency or absent in peripheral blood (PB). To address this, we evaluated the use of monocyte-derived macrophages (MØ) and dendritic cells (mo-DC) as screening tools to detect IIRMIs. Using purified PRR Agonists (PRRAgs) as model IIRMI and human IL-6 and IL-8 mRNA and protein as biomarkers for cell activation, we show that both primary cultures are more sensitive than PBMC in detecting IIRMI. In addition PRRAgs induced increased expression of a set of pro-inflammatory genes and the profile of genes induced varied with the TLR agonist and concentration. Finally, we tested the capability of human PBMC, MØ and mo-DC for detecting impurities in a commercially available product for the treatment of autoimmune inflammatory diseases. While PBMC samples were not activated by impurities present in the product, MØ and mo-DC showed a robust activation.