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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

Labor-intensive

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