Human PD1 Knockout Cell Line-Jurkat
Cat.No. : CSC-RT2315
Host Cell: Jurkat Target Gene: PD1
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT2315 |
| Cell Line Information | This cell is a stable cell line with a homozygous knockout of human PD1 using CRISPR/Cas9. |
| Target Gene | PD1 |
| Host Cell | Jurkat |
| Storage | Liquid nirtogen |
| Species | Human |
| 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. |
| 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 |
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Background
Case Study
Applications
The PD1 gene, also known as the programmed cell death protein 1 gene, is a key component in regulating immune responses. The PD1 gene is located on chromosome 2 in humans and encodes the PD1 protein, an immune checkpoint receptor expressed on the surface of T cells and pre-B cells. PD1 interacts with its ligands, PD-L1 and PD-L2, which are expressed on a variety of cells, including tumor cells, antigen-presenting cells, and other non-lymphocytes. Binding of PD1 to its ligands results in inhibition of T cell activation and proliferation. This interaction leads to T cell "exhaustion," which reduces their ability to effectively target and destroy cancer cells. As a result, many cancers exploit this pathway to evade the immune system, making PD1 an important focus of cancer immunotherapy research.
Checkpoint inhibitors targeting the PD1 pathway have revolutionized cancer treatment. Drugs such as pembrolizumab (Keytruda) and nivolumab (Opdivo) are monoclonal antibodies that block PD1, thereby reactivating T cells to recognize and attack tumor cells. These therapies have shown remarkable success in treating a variety of cancers, including melanoma, non-small cell lung cancer, and Hodgkin lymphoma.
Staphylococcus aureus (S. aureus) presents a significant challenge in the clinic due to its well-defined ability to select mutations that increase antibiotic resistance and immune evasion. However, the molecular mechanisms remain poorly understood, especially for adaptive immunity. Cancer immunotherapy targeting programmed cell death protein 1 (PD-1) enhances T cell activity and is being used to treat certain viral infections, whereas its potential against bacterial infections remains elusive. Here researchers show that S. aureus clpP mutants selected during clinical antibiotic therapy suppress T cell activity by directly interacting with PD-1 on human T cells. The specificity of the interaction was confirmed using recombinant PD-1 as well as PD-1 overexpressing and knockout cells. Furthermore, PD-1 binding to S. aureus inhibited intracellular calcium mobilization, T cell proliferation, CD25 expression, and IL-2 secretion, whereas antibody-mediated PD-1 blockade using an engineered IgG1-based anti-PD-1 antibody mitigated the major effects. These results suggest that clpP mutant S. aureus directly targets PD-1 to evade immune activation and that therapeutic targeting of PD-1 could be useful against certain staphylococcal infections.
Here, the researchers used a panel of five (SADR-1, SADR-2, SADR-3, SADR-4, and SADR-5) methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from the same patient during persistent bacteremia. Using fluorescently labeled S. aureus, they found that SADR-2, SADR-3, and SADR-4 interacted significantly more with Jurkat-PD1 than with Jurkat-WT, whereas SADR-1, SADR-5, and USA300JE2 showed significantly lower PD-1-specific interactions (Figure 1a). The interaction of SADR-2 with Jurkat-PD1 was verified by confocal microscopy (Figure 1b). Jurkat-PD1, Jurkat-WT, and Jurkat-PD1 knockout (KO) T cells were treated with PBS or AF647-labeled SADR-1 or SADR-2. Interactions were analyzed by flow cytometry 2 hours later (Figure 1c). The study showed that knockout (KO) of PD-1 greatly reduced the interaction of SADR-2 with Jurkat T cells, further confirming that PD-1 is essential for binding to SADR-2 (Figure 1c). In fact, the binding of the clpP mutant SADR-2 to IgG-fc was significantly increased, while the interaction between PD-1-Fc and SADR-1 was significantly reduced (Figure 1d). As expected, no interaction was observed with the control chimeric protein CD44-Fc (Figure 1d). These data strongly suggest that the suppressive phenotype of T cells exposed to SADR-2 is caused by a direct inhibitory interaction with PD-1.
Figure 1. ClpP mutant S. aureus interacts directly with PD-1. (Mellergaard M, et al. 2023)
Programmed cell death protein 1 (PD-1) is an immune checkpoint receptor expressed on T cells that plays a key role in regulating immune responses and maintaining self-tolerance by inhibiting T cell inflammatory activity. Here are some key applications of the Human PD-1 Knockout Cell Line - Jurkat:
Immunotherapy Research: The Human PD1 Knockout Cell Line - Jurkat is valuable for studying the immune checkpoint pathway, specifically the role of PD-1 in T cell regulation. This can aid in the development of novel immunotherapy strategies designed to enhance T cell activity against tumors.
Drug Screening: These cell lines are used to screen for potential drugs targeting the PD-1 pathway. By observing immune responses in the absence of PD-1, researchers can identify compounds that may inhibit or enhance immune activity, facilitating the discovery of effective therapies.
Cancer Research: Researchers can explore how tumor cells interact with T cells that lack PD-1 and develop strategies to prevent tumors from exploiting this checkpoint to avoid immune detection.
Basic Immunology Research: PD-1 knockout Jurkat cells provide insights into fundamental aspects of T cell receptor signaling and function. This understanding is critical to elucidating the broader role of immune checkpoints in immune system regulation.
Vaccine Development: By studying immune responses in a PD-1 deficient setting, researchers can gain insight into how to enhance vaccine efficacy. This is particularly important for designing vaccines designed to stimulate robust T cell responses.
For research use only. Not intended for any clinical use.
