Dendritic Cell Activation Assay

Why We Study Dendritic Cell Activation?

Aggregates in biotherapeutic drugs have been linked to the development of anti-drug antibodies (ADAs) in patients, prompting extensive research to unravel the cellular mechanisms that drive immunogenicity. Particularly, it was found that IgG aggregates can trigger the maturation of monocyte-derived dendritic cells (moDCs), evident through the elevated expression of maturation markers and the secretion of pro-inflammatory cytokines. Recent studies highlighted the potential of using DCs as models to evaluate the immunogenic possibility of therapeutic antibodies and their aggregates.

Fig.1 PAMPs-driven cDCs activation enables T cell priming.¹

Dendritic Cell Activation Assay at Creative Biolabs

To overcome the obstacles of biotherapeutics development, Creative Biolabs delivers a dendritic cell activation assay for clients around the world.

Our dendritic cell activation assay offers a robust in vitro platform utilizing moDCs for evaluating the immunogenic potential of diverse therapeutic agents through a high-throughput, multi-endpoint approach. We provide access to a comprehensive array of HLA-typed moDCs sourced from the blood of healthy human donors, enhancing the reliability of our results by capturing both individual and population-wide immune responses. This assay is also tailored to assess vaccines and adjuvants in the aspect of safety and efficacy. At Creative Biolabs, our seasoned experts are adept at customizing experimental designs to meet the specific needs of your projects, ensuring precise and relevant insights.

Fig.2 Workflow of our assay.

Multiple Endpoints Empower Our Dendritic Cell Activation Assay

Common DC Activation Markers at Creative Biolabs

• Cell surface markers: CD80, CD86, CD54, MHC I (HLA-ABC), MHC II (HLA- DR), CD83, CD25, and CD40
• Released cytokines and chemokines: MIP1β, IL-12, IL-23, IFN-γ, IL-1β, IL-6, IL-8, IL-12, TNFα, CCL3, and CCL4
• Intracellular signaling proteins: Syk, Akt, and ERK1/2.
• Cell viability: Cellular ATP

Key Advantages

• Decrease in reliance on animal testing and quicker evaluation of a candidate's suitability.
• Convenient endpoint reading.
• State-of-art technology platforms.
• Selective measurable indicators are available.
• Outstanding accuracy and consistency.
• One-step customized services.

Published Data

This study aims to develop predictive in vitro assays to monitor dendritic cell (DC) maturation and evaluate the direct stimulatory effects of drug products on DCs. By treating monocyte-derived DCs (moDCs) with four different antibody drugs in both native and aggregated forms, the findings reveal that DC activation assays are effective tools for screening therapeutic antibodies during their development stage to identify their inherent properties, including any formulation-induced aggregation, which might elevate the risk of triggering an immune response.

Fig.3 Phenotypic assessment of maturation markers on moDCs post-treatment with native or aggregated antibody drugs.²

Fig.4 Proinflammatory cytokine mRNA levels in moDC post-treatment with native or aggregated antibody drugs.²

Fig.5 Phosphorylation profiling of moDCs exposed to native versus aggregated antibody drugs.²

Q & A

1. Q: What are the concentrate requirements for candidate compounds?

   A: Provide up to 100 μL of a solution in PBS for a 20X final concentration or an equivalent amount of the solid compound. For compounds soluble in DMSO, provide at 200X concentration to ensure cultures remain at 0.5% v/v DMSO. Before this, screen compounds for pyrogenic contamination before submission; in-house pyrogenic testing is available at Creative Biolabs upon prior request if required.

If you are looking for a cost-effective dendritic cell activation assay to evaluate your compounds, please click the link or submit an inquiry to reach out to us.

References

1. Bourque, Jessica, and Daniel Hawiger. "Activation, amplification, and ablation as dynamic mechanisms of dendritic cell maturation." Biology 12.5 (2023): 716.
   Distributed under Open Access License CC BY 4.0, without modification.
2. Morgan, Hannah, et al. "Evaluation of in vitro assays to assess the modulation of dendritic cells functions by therapeutic antibodies and aggregates." Frontiers in Immunology 10 (2019): 601
   Distributed under Open Access License CC BY 4.0. The original images were not modified.

For Research Use Only.