Back to Search
Start Over
In vitro immunotherapy potency assays using real-time cell analysis
- Source :
- PLoS ONE, PLoS ONE, Vol 13, Iss 3, p e0193498 (2018)
- Publication Year :
- 2018
- Publisher :
- Public Library of Science, 2018.
-
Abstract
- A growing understanding of the molecular interactions between immune effector cells and target tumor cells, coupled with refined gene therapy approaches, are giving rise to novel cancer immunotherapeutics with remarkable efficacy in the clinic against both solid and liquid tumors. While immunotherapy holds tremendous promise for treatment of certain cancers, significant challenges remain in the clinical translation to many other types of cancers and also in minimizing adverse effects. Therefore, there is an urgent need for functional potency assays, in vitro and in vivo, that could model the complex interaction of immune cells with tumor cells and can be used to rapidly test the efficacy of different immunotherapy approaches, whether it is small molecule, biologics, cell therapies or combinations thereof. Herein we report the development of an xCELLigence real-time cytolytic in vitro potency assay that uses cellular impedance to continuously monitor the viability of target tumor cells while they are being subjected to different types of treatments. Specialized microtiter plates containing integrated gold microelectrodes enable the number, size, and surface attachment strength of adherent target tumor cells to be selectively monitored within a heterogeneous mixture that includes effector cells, antibodies, small molecules, etc. Through surface-tethering approach, the killing of liquid cancers can also be monitored. Using NK92 effector cells as example, results from RTCA potency assay are very well correlated with end point data from image-based assays as well as flow cytometry. Several effector cells, i.e., PBMC, NK, CAR-T were tested and validated as well as biological molecules such as Bi-specific T cell Engagers (BiTEs) targeting the EpCAM protein expressed on tumor cells and blocking antibodies against the immune checkpoint inhibitor PD-1. Using the specifically designed xCELLigence immunotherapy software, quantitative parameters such as KT50 (the amount of time it takes to kill 50% of the target tumor cells) and % cytolysis are calculated and used for comparing the relative efficacy of different reagents. In summary, our results demonstrate the xCELLigence platform to be well suited for potency assays, providing quantitative assessment with high reproducibility and a greatly simplified work flow.
- Subjects :
- 0301 basic medicine
Physiology
medicine.medical_treatment
Cell Lines
Programmed Cell Death 1 Receptor
Cancer Treatment
lcsh:Medicine
Apoptosis
Biochemistry
White Blood Cells
0302 clinical medicine
Spectrum Analysis Techniques
Cancer immunotherapy
Animal Cells
Immune Physiology
Medicine and Health Sciences
lcsh:Science
Multidisciplinary
Immune System Proteins
medicine.diagnostic_test
Cell Death
Chemistry
Effector
T Cells
Flow Cytometry
Epithelial Cell Adhesion Molecule
medicine.anatomical_structure
Oncology
Cell Processes
Spectrophotometry
030220 oncology & carcinogenesis
MCF-7 Cells
Biological Assay
Immunotherapy
Cytophotometry
Biological Cultures
Cellular Types
Raji Cells
Research Article
T cell
Immune Cells
Immunology
Cytological Techniques
Research and Analysis Methods
Cancer Immunotherapy
Antibodies
Flow cytometry
03 medical and health sciences
Immune system
In vivo
Cell Line, Tumor
medicine
Potency
Humans
Blood Cells
Cytolysis
lcsh:R
Biology and Life Sciences
Proteins
Cell Biology
030104 developmental biology
Cancer research
Leukocytes, Mononuclear
lcsh:Q
Clinical Immunology
Clinical Medicine
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 13
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....2636df993a85fb4ca6281efd2ce332ee