CLS-logo.png Concept Life Sciences has a team of immunology experts that helps its clients understand their targets' and compounds' MOA (mechanism of action) and immunomodulatory effects to progress their drug discovery pipeline.

We offer collaborative experimental design and can advise on the most suitable cell types, mono- or co-culture assays. By customizing and developing assays to suit your requirements, we can build a data package that is right for you.

We provide data you can trust with expert interpretation of the results, giving you greater confidence in your lead compound selection and accelerating your route to clinic.

Peripheral Blood Mononuclear Cells (PBMCs)

We take the time to understand your target pathway to offer the most appropriate assay for your stage of development e.g. screening for compound selection or detailed mechanism of action (MOA)/validation study.

PBMC assays are ideal if you require large scale analyses using simple immune stimuli. We can customize and design a PBMC study to meet your research needs e.g. TLR-stimulation for innate mononuclear cell activity or T cell receptor ligation for T cell function, accelerating your drug discovery pipeline.

PBMCs can be used in all areas of immunology, from autoimmunity to cancer, and in multiple assays. The data derived is informative for progressing to more advanced immunoassays (monocultures and/or co-cultures) designed to further interrogate the MOA of your compound.

T Cells

Understanding how T cells respond to your drug candidate is key to bringing your treatment to market.

We offer a variety of T cell assays to help you study the nuances of T cell interaction with your drug candidate. Whether you are developing anti-inflammatory or anti-cancer therapies, our comprehensive panel of T cell assays will provide you with valuable efficacy data.

T cell assays can focus on individual T cell populations, allowing you to ask more targeted research questions, complementing data obtained from more complex, mixed cell populations (e.g. PBMCs). Our T cell polarisation assays are excellent tools to study your compounds’ effects on differentiation and function of T cells, and the pathogenesis of various inflammatory diseases e.g. autoimmune and allergic diseases. We have a suite of human Th1, Th2, Th17 and Treg cell assays with standard and custom readouts to suit your study requirements.

Whatever your research needs, we can tailor a package to suit you. Alone or in combination with other cells, we offer:

  • T cell polarisation assay
  • T cell stimulation assay
  • T cell killing assay
  • Treg suppression assay
  • Antigen-specific T cell assay
  • Mixed lymphocyte reaction (MLR)

Natural killer (NK) cells

We provide tailored assays to study the effects of your drug candidate on NK cells and the wider immune response.

Because of their ability to kill tumor cells, NK cells are an attractive target for cancer immunotherapies especially therapeutic monoclonal antibodies, which act through antibody-mediated cellular cytotoxicity (ADCC).

Whether you’re investigating target antigen expression on tumor cells/cell lines or by immune suppressive cells such as regulatory T cells, M2 macrophages or myeloid-derived suppressor cells (MDSC) to drive an ADCC reaction, or evaluating NK cell cytotoxicity function, we can design a custom program around NK cells that specifically meets your research needs. Our scientific team can guide you in answering research questions to understand your molecule’s MOA, efficacy or aid in lead optimization.

We also offer custom and standard assay readouts such as cytokine expression by ELISA or multiplex, proliferation and cell surface markers using FACS, and live cell imaging using the IncuCyte Live-Cell Analysis System.

Dendritic cells (DCs)

We can help you understand the role of dendritic cells (DCs) and your drug candidate or target on the immune response.

We can screen DCs in a monoculture or in combination with T cells in a mixed lymphocyte reaction (allogeneic/autologous, immature/mature). We can offer custom assay readouts, in addition to standard flow cytometry (e.g. MHC, CD80, CD86, PD-L1) as well as cytokine release assays (e.g. IL-10 versus IL-12 from DCs or IL-10 versus IFN-γ from T cells).

Whether you’re studying cancer or autoimmune disease, we can tailor a study around DCs that meets your specific research needs. Our expert staff can guide you in answering research questions around DC maturation, polarisation, interaction, and efficacy as antigen-presenting cells.

Myeloid cells

Myeloid cells are the most abundant nucleated hematopoietic cells in the body, consisting of a range of cell types with diverse functions. They include monocytes, macrophages, DCs and granulocytes, and make up a critical arm of the immune system.

Our immunology experts can help you understand if your compound or biologic affects the phenotype or function of myeloid cells, in an isolated cell typesetting, or in a co-culture environment, ensuring the most appropriate level of complexity for you immunology discovery program.

Our monocyte and macrophage monocultures are ideal for pre-clinical optimization (e.g. determining dose response) without expensive animal testing. M1 macrophage assays are designed for studying compounds that inhibit pro-inflammatory responses in autoimmune diseases, such as rheumatoid arthritis. M2 macrophage assays are best suited for drug candidates designed to reverse the suppression of the immune system by the tumor microenvironment.

Dendritic cells, especially inflammatory DCs, arise from circulating monocytes. We can help you understand if your drug candidate impacts this differentiation process. Alternatively, we can support your investigations into your compound effect on the maturation of DCs.

For clients beginning in vivo studies or clinical trials, we offer functional assays for confirming drug effectiveness. 

Hepatocytes

Drug-induced liver injury (DILI) is one of the main causes of investigational new drug (IND) failure in clinic. At Concept Life Sciences, we understand the importance of knowing the effects your test compounds or biologics have on hepatocytes early in discovery.

Hepatocytes play an essential role in toxin and drug metabolism and clearance. We use primary hepatocytes (human, rat or mouse) in our in vitro assays to predict drug hepatotoxicity. Our primary cell assay systems evaluate the impact of your drug candidate on hepatocyte proliferation, metabolism and enzyme activity.

Keratinocytes

Keratinocytes are an important but challenging primary cell type to investigate. We know the importance of understanding the impact of your novel therapeutic candidate on keratinocyte function and their interaction with immune cells to advancing your discovery program.

Our customizable in vitro skin assays can evaluate keratinocyte proliferation, migration, differentiation, and cytokine production. We can source human primary keratinocytes from healthy and diseased patients such as Psoriasis and Atopic Dermatitis as well as various cell lines to provide translational models.

The skin is the largest organ in the body and an important therapeutic target for many diseases.

Cytotoxicity

Drug-induced toxicity is the leading cause of withdrawal of approved drugs from the market. Identifying potential tissue- or cell-relevant toxicity in the early stages of discovery will save you time and money, ultimately accelerating your route to clinic.

We understand your reasons for investigating cytotoxicity. Whether you need to screen your compound library to find selective cytotoxic IO drug candidates to take forward, or screen, your “hits” for unwanted cytotoxic effects, our immunology experts work closely with you to determine if your compound impacts the viability of both immune and non-immune cells, including tumor cells.

Using our primary cell know-how and custom in vitro assay approach, we can assess apoptosis and necrosis of mixed or isolated cell populations such as PBMCs, T cells, B cells, NK cells, monocytes and DCs in mono- or co-culture set up, with both standard and custom readouts e.g. IncuCyte Live-Cell Analysis System, flow cytometry, ELISA and qPCR.

We also offer Killing Assays to investigate targeted cell death mechanisms.

Migration assays

Cell migration is a key mechanism of live cells and vital for an immune response and other disease processes. Assays to evaluate cell migration are important in understanding cancer metastasis and inflammation as well as wound healing.

We have a suite of in vitro cell migration assays such as the scratch wound and transwell migration assays to study live cell behavior in response to your novel drug candidates.

Scratch wound migration assay

A simple wound-healing assay to study directional cell migration in vitro using various cell lines chosen for your requirements. A “wound” is generated on a confluent cell monolayer and detailed cell migratory behavior is evaluated using time-lapse microscopic imaging. Further customizable readouts include functional cell changes by FACS and ELISA.

Transwell migration assay

Investigate compound effects on immune cell migration with the transwell migration assay.

Cells are seeded in the upper compartment of the transwell and stimulated with a chemotactic reagent in the lower compartment. Cell migration is monitored over time, and migrated cells are collected and enumerated from the lower compartment using FACS and counting beads. We can customize this assay by using specific immune cells and chemoattractants.

Immune safety assays

Concept Life Sciences offers a science-led optimization approach for your test substance in determining the most suitable cytokine release assay (CRA) format before performing a screen in a larger donor cohort. 

New molecular entities may induce infusion reactions and in turn, lead to cytokine release syndrome (CRS) characterized by a broad inflammatory response and adverse reactions.

Following the TeGenero TGN1412 FIH trial that led to 6 volunteers suffering multi-organ failure1, the FDA issued draft guidance on Nonclinical Safety Evaluation of the Immunotoxic Potential of Drugs and Biologics (Feb 2020).

There are multiple protocols to test Human cell-based in vitro assays to detect harmful cytokine release and each needs optimizing for each Test Substance2.

For more detail on our CRA assay (Figure 1 How our cytokine release assay is performed) please download our assay flier.

References:

  1. BMJ 2006;332:677 - Learning from the TGN1412 trial 
  2. Findlay, Eastwood, Ball et al. 2011; Comparison of novel methods for predicting the risk of pro-inflammatory clinical infusion reactions during monoclonal antibody therapy. Journal of Immunological Methods, 371, 134-142

How our cytokine release assay is performed (Figure 1)

1. Peripheral blood samples from multiple donors are used to ensure genetic diversity.2. Test and control compounds are prepared to required concentrations and applied wet coated onto assay plates or used in aqueous phase; diluted blood is then added and plates are incubated for 1, 6 or 24 hrs.

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3. Test substances interact with their specific cellular targets leading to the release of cytokines from activated immune cells.4. Assay plates are spun down and supernatants are used to measure a panel of cytokines such as GMCSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, and TNFα using the MSD technology.

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5. Data are analyzed and displayed in a series of convenient formats such as heatmaps, bar graphs and dot plots to provide a quick overview of cytokine responses across different donors or to determine the population of responders vs non-responders for individual treatments and cytokine. CRA flow image 5.png

Inflammatory in vivo models

We understand investigating the MOA of your compound using an in vivo model is a requirement on your route to clinic. We have a range of in vivo and ex vivo platforms that can be leveraged to study the effect of your drug candidate in an inflammatory environment.

We work closely with you to select the most suitable approach based on the immune cells or disease of interest to better understand your compound or biologic.

Our customizable models can study but are not limited to the following:

  • Antigenic challenge
  • Autoimmune inflammation of the CNS (EAE)
  • Delayed-type hypersensitivity (DTH)
  • Macrophage biology
  • Liver disease

We offer standard and tailored readouts ensuring robust MOA interrogation of your drug candidate. Our histology experts offer standard H&E for histological analysis through to specialist multiplex immunofluorescent staining, and our immunology expertise allows for tailored FACS analysis, ELISA, qPCR and NGS readouts.