Cat.No. : CSC-RO0518 Host Cell: HEK293T
| Cat. No. | CSC-RO0518 |
| Description | This cell line is derived from HEK293T and is engineered to stably overexpress Human CD27. |
| Gene | CD27 |
| Gene Species | Homo sapiens (Human) |
| Host Cell | HEK293T |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Studying the interactions between immune cells and cancer cells 2. Studying the mechanisms of resistance to immune checkpoint blockade 3. High-throughput screening 4. Drug target validation |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Growth Properties | Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2. Split at 80-90% confluence, approximately 1:3-1:6. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
CD27 is a co-stimulatory molecule that provides a complementary target for the PD-1/PD-L1 checkpoint axis on T cells. Here, researchers developed CDX-527, a tetravalent PD-L1xCD27 IgG1-scFv bispecific antibody (BsAb). CDX-527 potently inhibits PD-1 signaling and induces CD27-mediated T cell co-stimulation via PD-L1 cross-linking. In a mixed lymphocyte reaction assay, CDX-527 outperformed the combination of parental antibodies, suggesting that cross-linking via Fc receptors and PD-L1 enhances CD27 agonist activity. CDX-527 has been shown to mediate effector functions against tumor cells overexpressing CD27 or PD-L1. In human CD27 transgenic mice, researchers observed that antigen-specific T cell responses to vaccines were significantly enhanced using an alternative PD-L1xCD27 BsAb. In addition, in a syngeneic lymphoma model, this BsAb showed higher antitumor activity than the parental antibody combination. Preliminary studies of CDX-527 in cynomolgus monkeys confirmed its mAb-like pharmacokinetic characteristics without significant toxicity. These studies showed that CDX-527 effectively combines PD-1 blockade and CD27 co-stimulation into a single molecule with greater potency than the parental antibody combination, which provides a theoretical basis for advancing clinical research of this BsAb in cancer patients.
BsAbs are designed to enhance T cell immunity, and CDX-527 binds to activated FcγRs, potentially mediating effector functions such as ADCC against certain tumor cells that overexpress PD-L1 or CD27. CDX-527 induced ADCC in tumor cell lines endogenously expressing PD-L1 or CD27 or in cells transfected with these receptors, although the BsAb was slightly less potent than the parental 2B3 mAb in CD27-expressing cells (Figure 1a-d). To confirm target specificity, 9H9 induced ADCC only in PD-L1-expressing cells but not in CD27-expressing cells, while 2B3 induced ADCC in CD27-expressing cells but not in PD-L1-expressing cells. CDX-527 did not exhibit measurable complement-mediated cytotoxicity against the same cell lines.
Figure 1. CDX-527 mediated ADCC. (Vitale L A, et al., 2020)
A: The Human CD27 Stable Cell Line - HEK293T, with stable expression of the CD27 receptor, holds significant potential for tumor immunotherapy research. CD27 is a crucial immunoregulatory receptor, and its activation can enhance the immune responses of T cells and B cells. This cell line can be used to study the role of CD27 in tumor immune evasion and anti-tumor immune responses.
A: By using the Human CD27 Stable Cell Line - HEK293T, various biochemical and molecular biology experiments can be conducted to study the regulatory mechanisms of the CD27 signaling pathway. For example, immunoprecipitation and mass spectrometry can be used to identify proteins interacting with CD27, and reporter gene analysis can monitor the activity of the CD27 signaling pathway.
A: For antibody screening, experiments should be designed to assess the binding capacity and functional impact of different candidate antibodies on the CD27 receptor in the Human CD27 Stable Cell Line - HEK293T. Techniques like flow cytometry or ELISA can be used to evaluate the binding affinity and antigen specificity of antibodies.
A: The Human CD27 Stable Cell Line - HEK293T can be used to simulate a chronic inflammation environment to study the role and regulatory mechanisms of CD27 in chronic inflammation. By simulating chronic inflammatory conditions, the effects of CD27 on immune cell activation, cytokine production, and cell migration can be studied.
A: When conducting gene editing experiments, it is important to select appropriate targets and use efficient and specific gene editing technologies, such as the CRISPR/Cas9 system. Detailed molecular and cellular level validations are needed to ensure the accuracy of gene editing and its impact on CD27 expression and function.
A: The Human CD27 Stable Cell Line - HEK293T has potential applications in the study of autoimmune diseases. CD27 plays a crucial role in regulating the activation and differentiation of T cells and B cells, and this cell line can be used to investigate the mechanisms of CD27 in autoimmune diseases and potential therapeutic targets.
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