scFv(EPCAM)-CD28-CD3zeta CAR-T Lentivirus

scFv(EPCAM)-CD28-CD3zeta CAR-T lentivirus is a highly advanced and efficient T-lymphocyte gene-engineering vector that specifically targets epithelial cell adhesion molecule (EpCAM). This lentiviral vector system exhibits excellent transduction efficiency and can stably and permanently integrate transgenes into the host cell genome. Unlike retroviral vectors, lentiviruses can infect both dividing and non-dividing cells, making them ideal for modifying primary human T cells with different proliferative states. The vector is typically pseudotyped with vesicular stomatitis virus glycoprotein (VSV-G), which imparts broad tropism and enhances its physical stability during concentration and purification. A key advantage of this construct lies in its second-generation chimeric antigen receptor (CAR) structure. By integrating the CD28 co-stimulatory domain into the CD3-ζ primary signaling chain, the lentivirus ensures that the generated CAR-T cells receive the "second signal" required for full activation. Compared to first-generation CARs, this design significantly enhances T cell proliferation, cytokine production (such as IL-2 and IFN-γ), and anti-apoptotic capabilities.

The scFv(EPCAM)-CD28-CD3ζ CAR-T lentivirus has a wide range of applications, primarily focused on the rapidly developing field of cancer immunotherapy. EpCAM is a well-known transmembrane glycoprotein that is significantly overexpressed in various cancer tissues, including colorectal cancer, breast cancer, lung cancer, prostate cancer, and ovarian cancer, while its expression is limited in normal epithelial tissues. In the laboratory, these lentiviral particles are used to generate CAR-T cells for in vitro cytotoxicity assays. In these assays, engineered T cells are co-cultured with EpCAM-positive tumor cell lines to assess their antigen-specific killing, degranulation, and metabolic adaptation. Beyond basic research, this lentivirus is also crucial for in vivo preclinical studies; researchers have used it to construct CAR-T cell populations and infused them into xenograft mouse models to monitor tumor regression, CAR-T cell persistence, and infiltration into the tumor microenvironment. Furthermore, this system can serve as a benchmark for testing novel modifications in CAR designs, such as adding "safety switches" or adjusting the hinge and transmembrane regions to optimize hinge flexibility. It also plays a crucial role in drug screening and combination therapy research, as CAR-T cells generated from this lentivirus can be used in combination with immune checkpoint inhibitors or small molecule drugs to overcome the immunosuppressive barrier of the solid tumor microenvironment.