scFv(GPC3)-OX40-CD3zeta CAR-T Lentivirus

The scFv(GPC3)-OX40-CD3zeta CAR-T lentivirus is a tool for cell immunotherapy research, employing a sophisticated delivery system designed to achieve maximum transduction efficiency and long-term stability. One of the key features of this lentiviral vector is its ability to permanently integrate the anti-GPC3 CAR transgene into the host cell genome. This genomic integration ensures that the chimeric antigen receptor (CAR) is inherited by all daughter cells during amplification or T cell proliferation in the patient, resulting in sustained and stable expression of the therapeutic protein. Furthermore, the lentiviral delivery system can transduce both dividing and non-dividing cells, a significant improvement over traditional retroviral vectors. These lentiviral particles are typically pseudotyped with the vesicular stomatitis virus G glycoprotein (VSV-G), conferring broad tropism and enabling efficient and low-toxicity entry into T lymphocytes. The vector''s safety is also a major highlight; the deletion of the U3 region in the 3'' long terminal repeat (LTR) prevents the regeneration of replicating lentiviruses, making it a safe choice for preclinical and clinical research settings.

The primary application of the scFv(GPC3)-OX40-CD3zeta CAR-T lentivirus is the development and evaluation of targeted therapies against hepatocellular carcinoma (HCC) and other solid tumors overexpressing Glypican-3 (GPC3). GPC3 is a carcinoembryonic antigen highly expressed on the surface of HCC cells but almost absent in healthy adult tissues, making it an ideal target for reducing off-target toxicity. This particular second-generation CAR construct contains a single-chain variable region fragment (scFv) derived from a high-affinity anti-GPC3 antibody, linked with an OX40 (CD134) co-stimulatory domain and a CD3ζ activation tail. While many CAR-T cell designs utilize CD28 or 4-1BB, the inclusion of OX40 provides unique biological advantages, including enhanced T cell survival, increased production of pro-inflammatory cytokines such as IL-2 and IFN-γ, and promotion of memory T cell phenotype. These lentiviral particles are used to generate CAR-T cells for in vitro cytotoxicity assays to detect their precise killing effect on GPC3-positive tumor cells. Furthermore, they are crucial for in vivo studies using xenograft or orthotopic mouse models, which can assess the ability of engineered cells to infiltrate solid tumors and overcome the immunosuppressive tumor microenvironment.