Novel, Highly Sensitive, Non-Radioactive Cytotoxicity Assay with Cytation 5
Institut de Pharmacologie Moléculaire et Cellulaire (IPMC)
At the Institut de Pharmacologie Moléculaire et Cellulaire (IPMC) in Nice/Sophia-Antipolis, France, Drs. Fabienne Anjuère and Véronique Braud are on the cutting edge of immunotherapy research with the goals of identifying therapeutic targets and developing innovative vaccines and immunotherapies to treat inflammatory disorders and cancers. Most recently, they are focused on the study of innate immune cells (natural killer cells, neutrophils, dendritic cells) in healthy and tumorigenic epithelial tissues, which requires use of flow cytometry, cellular and biochemical assays in both in vitro and in vivo studies.
Their goal was to develop a functional assay to measure cellular cytotoxicity in innate immune cells such as natural killer (NK) cells. The 51Chromium (51Cr) release assay is a commonly used and highly sensitive method for this purpose, however, Dr. Braud was concerned about the high costs and hazards associated with handling and disposing radioactive materials. The alternative commercial assays using non-radioactive materials didn’t provide the required sensitivity. Faced with the challenge of developing a novel cellular cytotoxicity assay that was both highly sensitive and non-radioactive, Dr. Braud set about looking for combination imaging and detection instruments.
As she considered instrumentation options, Dr. Braud discovered BioTek’s Cytation 5 Cell Imaging Multi-Mode Reader and the convenience of a combined cell imaging and multi-mode instrument. Cytation™ 5 could provide what she was looking for in terms of multiple excitation/emission wavelengths along with temperature and environmental control.
Using the Cytation 5, Dr. Braud successfully developed a novel, non-radioactive cytotoxicity assay with
sensitivity equivalent to the 51Cr assay along with real-time monitoring. The assay is capable of measuring NK and T cell cytotoxicity in the presence or absence of blocking antibodies as well as measuring antibody-dependent cell-mediated cytotoxicity (ADCC).
Knowing that the assay method would profoundly impact the scientific community, from academia through drug development and into the clinic, Dr. Braud published two papers. The first paper, titled, “A Real-Time Digital Bio-Imaging System to Quantify Cellular Cytotoxicity as an Alternative to the Standard Chromium-51 Release Assay” (DOI: 10.1111/imm.12702) was published in Immunology. The second paper, titled, “A Real-Time Cytotoxicity Assay as an Alternative to the Standard Chromium-51 Release Assay for Measurement of Human NK and T Cell Cytotoxic Activity” (DOI: 10.1002/cpim.28), was published in Current Protocols in Immunology, where it was recently cited as one of the Top 20 most downloaded recent papers in the Journal.
The Anjuère and Braud laboratory found that Cytation 5 provided many advantages beyond its utility in the novel cytotoxicity assay. Multiple imaging objectives and filter cubes expand the lab’s range of imaging applications, while multiple reading modes enable fast analysis of 96-well plates in numerous biochemical and cellular detection-based assays. Dr. Braud appreciates that a single instrument – the Cytation 5 – simultaneously expands and facilitates the array of technical applications performed in her laboratory.
Dr. Véronique Braud in her laboratory with BioTek’s Cytation 5 Cell Imaging Multi-Mode Reader
The Institut de Pharmacologie Moléculaire et Cellulaire (IPMC), located in Nice/Sophia-Antipolis, France.
To learn more about the Institut de Pharmacologie Moléculaire et Cellulaire (IPMC), visit their web site.
Thanks to Dr. Véronique Braud at IPMC for sharing her BioTek experience.