title: In Vivo Effect of Innate Immune Response Modulating Impurities on the Skin Milieu Using a Macaque Model: Impact on Product Immunogenicity.
abstract: Unwanted immune responses to therapeutic proteins can severely impact their safety and efficacy. Studies show that the presence of trace amounts of host cells and process-related impurities that stimulate pattern recognition receptors (PRR) can cause local inflammation and enhance product immunogenicity. Here we used purified PRR agonists as model impurities to assess the minimal level of individual innate immune response modulating impurities (IIRMIs) that could activate a local immune response. We show that levels of endotoxin as low as 10 pg (0.01 EU), 1 ng for polyinosinic:polycytidylic acid (PolyI:C), 100 ng for synthetic diacylated liopprotein, thiazoloquinolone compound, or muramyl dipeptide, 1 μg for flagellin or β-glucan, or 5 μg for CpG-oligodeoxynucleotide increased expression of genes linked to innate immune activation and inflammatory processes in the skin of rhesus macaques. Furthermore, spiking studies using rasburicase as a model therapeutic showed that the levels of PRR agonists that induced detectable gene upregulation in the skin were associated with increased immunogenicity for rasburicase. This study underscores the need for testing multiple IIRMIs in biologics, strengthening the connection between the local mRNA induction in skin, innate immune activation, and antibody development in primates, and provides an indication of the levels of IIRMI in therapeutic products that could impact product immunogenicity.
authors.affiliation: Laboratory of Immunology, Division of Biotechnology Review and Research III, Office of Biotechnology Products, Center for Drug Evaluation and Research, Food and Drug Administration (FDA), Silver Spring, Maryland 20993.
authors.forename: Lydia A
authors.initials: LA
authors.lastname: Haile
authors.affiliation: Laboratory of Immunology, Division of Biotechnology Review and Research III, Office of Biotechnology Products, Center for Drug Evaluation and Research, Food and Drug Administration (FDA), Silver Spring, Maryland 20993.
authors.forename: Montserrat
authors.initials: M
authors.lastname: Puig
authors.affiliation: Laboratory of Immunology, Division of Biotechnology Review and Research III, Office of Biotechnology Products, Center for Drug Evaluation and Research, Food and Drug Administration (FDA), Silver Spring, Maryland 20993.
authors.forename: Swamy K
authors.initials: SK
authors.lastname: Polumuri
authors.affiliation: Laboratory of Immunology, Division of Biotechnology Review and Research III, Office of Biotechnology Products, Center for Drug Evaluation and Research, Food and Drug Administration (FDA), Silver Spring, Maryland 20993.
authors.forename: Jill
authors.initials: J
authors.lastname: Ascher
authors.affiliation: Laboratory of Immunology, Division of Biotechnology Review and Research III, Office of Biotechnology Products, Center for Drug Evaluation and Research, Food and Drug Administration (FDA), Silver Spring, Maryland 20993. Electronic address: Daniela.Verthelyi@fda.hhs.gov.
authors.forename: Daniela
authors.initials: D
authors.lastname: Verthelyi
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D004731:Endotoxins / Q000008:administration & dosage / Q000276:immunology*; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects / Q000276:immunology*; D055261:Immunogenetic Phenomena / Q000187:drug effects / Q000502:physiology*; D007279:Injections, Subcutaneous; D008253:Macaca mulatta; D008297:Male; D023421:Models, Animal; D011066:Poly C / Q000008:administration & dosage / Q000276:immunology; D051192:Receptors, Pattern Recognition / Q000276:immunology; D012867:Skin / Q000187:drug effects / Q000276:immunology*
chemical_list: D000906:Antibodies; D004731:Endotoxins; D051192:Receptors, Pattern Recognition; D011066:Poly C
keywords: biosimilar; biotechnology; immune response; immunology; injectables; skin