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Table 1 Comparison of currently available methods for evaluating CAR efficacy in research lab and in clinic

From: The Role of Immunological Synapse in Predicting the Efficacy of Chimeric Antigen Receptor (CAR) Immunotherapy

Method Types Currently available Methods Assay readouts Pros Cons
In vitro assays Immunophenotyping assay ratio of CD4 and CD8 Easy and quick Cannot be used a predictive biomarker due to variations from each individual.
Cannot reflect the phenotyping in vivo
Proliferation and Cytokine secretion assay IL-2 and IFN-gamma productions Easy and quick Cannot reflect the proliferation and cytokine productions in vivo
Cytotoxicity by standard 51Cr-release assay 4-hour killing of tumor cells Easy and quick Short -term in vitro activation
No interaction with host real tumor cells
Long-term killing assay Number of CAR-T cells Reflects the CAR-T expansion and tumor killing in 1- or 2-week in vitro assay Time-consuming
Number of artificial tumor cells Artificial modified tumor cell lines
Slow and variable
Technically complex
Available in vitro Strategies for clinical use CAR-T cells Vector copy number (VCN) and CAR expression Technically convenient Cannot be used as predictive biomarker due to variations from each individual.
In vivo assays NSG mouse model Tumor size and tumor growth Predicts persistence of CAR cells No host immune system
Mouse survival Predicts CAR-T killing capability No tumor microenvironment (TME)
Body weight Easy to use Expensive
Syngeneic transplantable model Tumor size Intact host immune system Slow and complex
Mouse survival Some TME development Model is variable
Body weight Expensive
Biodistribution of CAR cells Labor-intensive