Oncology Assays
Immunogenic Cell Death
This is, as the name suggests an immune driven process. Immunogenic cell death is driven by stress, production of reactive oxygen species and increases in Damage Associated Molecular Patterns (DAMPs). These DAMPs include Heat Shock Proteins (HSP), High Mobility Group Box 1 (HMGB1), Calreticulin and ATP. The ability of chemotherapeutic agents to increase these DAMPs increases the likelihood of immune killing.
Expression of HSP70, HSP90 in ovarian cancer cell lysates (OV90) after treatment with Doxorubicin. Data presented as a fold change when compared with untreated cells.
Concentration of HMGB1 in cell culture supernatants of OV-90 cells treated with test compounds. HMGB1 ELISA was performed in duplicate and mean values (±SD) plotted for each condition. Standard curve (left) of known HMGB1 concentrations was used to determine concentration of HMGB1 for each experimental sample (right).
Calreticulin surface expression of OV-90 cells by flow cytometry. Surface expression of calreticulin shown as fold change in MFI compared to untreated cells shown on the left. Percentage of cells positive for calreticulin displayed on the right. Mean values (±SD) of duplicates plotted for all graphs.
Amount of ATP in cell culture supernatants of OV-90 cells treated with test compounds. ATP was quantified using ATP determination kit (Invitrogen) and mean values (±SD) plotted for each condition. Standard curve of known ATP amounts was used to determine quantity of ATP for each experimental sample.
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FAQ
Immunogenic cell death (ICD) is a form of regulated cell death that stimulates an immune response against tumour antigens. Unlike non-immunogenic apoptosis, ICD enhances anti-tumour immunity and is highly relevant in immuno-oncology.
ICD is characterised by the release and exposure of damage-associated molecular patterns (DAMPs), including calreticulin, ATP, and HMGB1, which promote dendritic cell activation and antigen presentation. At Cellomatics, these markers are quantified using high-content imaging, flow cytometry, and multiplex assays to provide sensitive, reproducible assessment of immunogenic cell death.
Cellomatics uses high-content imaging and flow cytometry to quantify DAMP expression at the single-cell level, enabling sensitive and reproducible analysis.
At Cellomatics, ICD assays are performed in both 2D and 3D tumour models and can be combined with immune cell co-cultures to assess downstream immune activation. This is further supported using primary human immune cells and advanced co-culture systems, enabling physiologically relevant evaluation of tumour–immune interactions and immunogenic responses.
Cellomatics offers expertise in immunology and bespoke assay development, delivering detailed mechanistic data that supports immuno-oncology programme advancement.