ABSTRACT
OBJECTIVE: We aimed to assess the role of interleukin - 1 receptor antagonist (IL-1RA) in a ligature-induced periodontal (LIP) model and the mechanism of IL-1RA in regulating the IL-17-mediated periodontal bone loss. DESIGN: Periodontal bone loss was induced through the LIP model in WT and Il1ra-/- mice and measured by micro(µ) CT. Transcription of upstream IL-17 production signals and downstream targets in the ligated gingiva was compared by the real-time quantitative PCR (RT-qPCR) between WT and Il1ra-/- mice. Single-cell suspensions were prepared in gingiva and cervical lymph nodes and were analyzed by fluorescence-activated cell sorting to quantify IL-17+ cells and IL-17-secreting subpopulations. We locally delivered an anti-IL-17 neutralizing antibody to the ligated gingiva and compared the bone loss with the isotype control antibody-treated Il1ra-/- mice. RESULTS: Il1ra-/- mice manifested significantly more bone loss than that of WT mice in the LIP model. Il17 and IL-17-associated transcripts (Il1b, Il6, Il23, Tgfb), Inos, Mrc1, Mmp13, and Rank were upregulated in the gingiva of Il1ra-/- mice in comparison to WT mice. Significantly more IL-17+ immune cells (CD45+IL17+) are present in the gingiva of Il1ra-/- mice with the majority of being TCR γδ T cells (CD45+IL-17+CD3+TCR γδ+) than WT mice. The anti-IL-17 neutralizing antibody treatment attenuated the alveolar bone loss in the LIP model. CONCLUSION: IL-1RA plays a protective role in the murine LIP model by suppressing an expansion of the IL-17+ cells and preventing a hyper-IL-17 response in the gingiva.
Subject(s)
Alveolar Bone Loss , Bone Diseases, Metabolic , Periodontitis , Animals , Mice , Alveolar Bone Loss/prevention & control , Alveolar Bone Loss/pathology , Antibodies, Neutralizing/pharmacology , Interleukin 1 Receptor Antagonist Protein/pharmacology , Periodontitis/drug therapy , Periodontitis/pathology , Receptors, Antigen, T-Cell , Receptors, Interleukin-1 , Th17 CellsABSTRACT
In characterizing a cell culture process to support regulatory activities such as process validation and Quality by Design, developing a representative scale down model for design space definition is of great importance. The manufacturing bioreactor should ideally reproduce bench scale performance with respect to all measurable parameters. However, due to intrinsic geometric differences between scales, process performance at manufacturing scale often varies from bench scale performance, typically exhibiting differences in parameters such as cell growth, protein productivity, and/or dissolved carbon dioxide concentration. Here, we describe a case study in which a bench scale cell culture process model is developed to mimic historical manufacturing scale performance for a late stage CHO-based monoclonal antibody program. Using multivariate analysis (MVA) as primary data analysis tool in addition to traditional univariate analysis techniques to identify gaps between scales, process adjustments were implemented at bench scale resulting in an improved scale down cell culture process model. Finally we propose an approach for small scale model qualification including three main aspects: MVA, comparison of key physiological rates, and comparison of product quality attributes.
