CD274 (PDL1) Chimeric Antigen Receptor (CAR): A Comprehensive Guide and Our Service & Product Introduction

CD274 (also known as PDL1, Programmed Death-Ligand 1) is a key immune checkpoint molecule that plays a crucial role in regulating immune responses and tumor immune escape. As a promising target in cancer immunotherapy, CD274 (PDL1) Chimeric Antigen Receptor (CAR) has attracted extensive attention in academic research and clinical trials.
With the continuous development of CD274 (PDL1) CAR research, the demand for high-quality plasmid vectors and professional customized services is increasing. RGBiotech is committed to providing high-quality CD274 (PDL1) CAR expression plasmid vectors and customized plasmid construction services, supporting global researchers and biotech companies in advancing CD274 (PDL1)-targeted immunotherapy research and development. If you are engaged in CD274 (PDL1) CAR research, whether you need standard plasmid vectors or customized construction services, we are your ideal partner. Contact us today at admin@rgbiotech.com to learn more about our products and services, and let us work together to promote the development of CD274 (PDL1)-targeted immunotherapy, bringing new hope to cancer patients.

Our CD274 (PDL1) CAR Expression Plasmid Vector Products and Custom Services

CD274 (PDL1) CAR is a cutting-edge technology in cancer immunotherapy, with broad application prospects in the treatment of various refractory tumors, including NSCLC, gastric cancer, urothelial carcinoma, and lymphoma.
RGBiotech is a professional provider of CD274 (PDL1) CAR expression plasmid vectors and customized plasmid construction services, dedicated to supporting global researchers and biotech companies in CD274 (PDL1)-targeted immunotherapy research and development. RGBiotech offers a full range of CD274 (PDL1) CAR expression plasmid vectors covering all five generations, with diverse vector backbones and rich functional features, meeting the needs of different research stages and application scenarios. We have rich experience in CD274 (PDL1) CAR plasmid vector development and customization. Our products are of high quality, stable performance, and have been widely used in academic research and industrial development, winning the trust of customers worldwide.

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Introduction of CD274 (PDL1)

CD274, officially named CD274 molecule, is a protein-coding gene with the HGNC ID 17635 and NCBI Gene ID 29126, updated on April 8, 2026. It is located on human chromosome 9p24.1, consists of 7 exons, and undergoes alternative splicing to produce multiple transcript variants, leading to different protein isoforms (e.g., isoform a and isoform b, with isoform b being shorter due to the lack of an in-frame exon in the 5' coding region). The CD274 gene is highly conserved across species, and its expression is tightly regulated at the genetic, epigenetic, transcriptional, post-transcriptional, and post-translational levels, with significant upregulation in response to inflammatory factors and tumor microenvironment stimuli. CD274 gene research is essential for understanding its role in tumor immune escape and developing targeted immunotherapies, including CD274 (PDL1) CAR-T therapy.

CD274 (PDL1) is a type I transmembrane glycoprotein belonging to the B7 family of costimulatory molecules, with a molecular weight of approximately 40-45 kDa. Its structure consists of four main parts: an extracellular region, a transmembrane domain, a short cytoplasmic tail, and a signal peptide. The extracellular region contains two immunoglobulin-like domains (IgV and IgC), which are responsible for binding to its cognate receptor PD-1 (Programmed Death-1, CD279) and other ligands such as B7-1 (CD80). The hydrophobic transmembrane domain anchors the protein to the cell membrane, while the 30-amino-acid cytoplasmic tail lacks obvious signaling motifs but participates in the regulation of protein stability and intracellular trafficking. The unique structure of CD274 (PDL1) makes it an ideal target for CAR-T cell therapy, as the extracellular IgV domain can be specifically recognized by the scFv of CD274 (PDL1) CAR.

The core function of CD274 (PDL1) is to regulate immune homeostasis by inhibiting T cell activation and cytokine production, thereby preventing autoimmunity and maintaining immune tolerance under normal physiological conditions. When CD274 (PDL1) binds to PD-1 on the surface of activated T cells, it transmits inhibitory signals that induce T cell exhaustion, apoptosis, or anergy, weakening the immune response to self-antigens or foreign antigens (e.g., pathogens). In the tumor microenvironment, CD274 (PDL1) is frequently overexpressed on tumor cells and tumor-infiltrating macrophages, enabling tumor cells to escape immune surveillance by inhibiting the cytotoxic activity of T cells and NK cells, which is a key mechanism of tumor immune escape. Additionally, CD274 (PDL1) is involved in regulating B cell activation, dendritic cell function, and memory T cell formation, and plays a role in tissue development, regeneration, and pain modulation. The overexpression of CD274 (PDL1) in tumors makes it a key target for CD274 (PDL1) CAR-T therapy, which can directly eliminate CD274 (PDL1)-positive tumor cells and reverse T cell exhaustion.

CD274 (PDL1) has a broad tissue distribution, with low basal expression in normal tissues and significantly upregulated expression in activated immune cells and tumor tissues. In normal tissues, it is weakly expressed in the appendix (RPKM 6.7), placenta (RPKM 4.5), and 23 other tissues, as well as on resting T cells, B cells, dendritic cells (DCs), and macrophages, with further upregulation upon cellular activation. It is also expressed on parenchymal cells such as vascular endothelial cells and pancreatic islet cells. In tumor tissues, CD274 (PDL1) is highly expressed in a variety of malignancies, including non-small cell lung carcinoma (NSCLC), gastric cancer, urothelial carcinoma, renal cell carcinoma (RCC), classical Hodgkin lymphoma (CHL), primary mediastinal large B-cell lymphoma, and EBV-associated malignancies. The high expression of CD274 (PDL1) in tumors and low expression in normal tissues ensures the specificity of CD274 (PDL1) CAR-T therapy, reducing off-target effects. This tissue distribution feature is crucial for the development of CD274 (PDL1) CAR expression plasmid vectors, as it guides the design of targeted and safe CAR constructs.

CD274 (PDL1) is closely associated with a variety of diseases, mainly including malignant tumors and autoimmune diseases, due to its role in immune regulation. In malignant tumors, CD274 (PDL1) overexpression is associated with poor prognosis, increased tumor invasiveness, and resistance to conventional chemotherapy and immunotherapy in many cancer types, such as NSCLC, gastric cancer, urothelial carcinoma, RCC, melanoma, and lymphomas. In autoimmune diseases (e.g., systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, type 1 diabetes), the dysregulation of CD274 (PDL1) expression disrupts immune tolerance, leading to abnormal activation of autoreactive T cells and tissue damage. Additionally, CD274 (PDL1) is involved in the regulation of transplant rejection and chronic infections, as its interaction with PD-1 can inhibit T cell activation and reduce immune rejection after organ transplantation. The close association between CD274 (PDL1) and various diseases highlights the broad application prospects of CD274 (PDL1) CAR technology in disease treatment, especially in refractory tumors.

Introduction of CD274 (PDL1) Chimeric Antigen Receptor (CAR)

CD274 (PDL1) CAR is a genetically engineered chimeric antigen receptor that specifically targets the CD274 (PDL1) protein on the surface of tumor cells and immune cells. Similar to other CARs, CD274 (PDL1) CAR consists of four core components: an extracellular single-chain variable fragment (scFv) that specifically binds to CD274 (PDL1), an extracellular hinge/spacer region that provides flexibility and stability, a transmembrane domain that anchors the CAR to the T cell membrane, and intracellular signaling domains that activate T cell cytotoxicity, proliferation, and persistence. By modifying T cells to express CD274 (PDL1) CAR, the engineered CAR-T cells can specifically recognize and bind to CD274 (PDL1)-positive cells, bypassing MHC restriction, and directly kill tumor cells or reverse T cell exhaustion, thereby enhancing anti-tumor immune responses. CD274 (PDL1) CAR is a novel immunotherapeutic strategy that complements traditional PD-1/PD-L1 antibody therapy, especially for patients with drug resistance to antibodies.

CD274 (PDL1) CAR has evolved through five generations, with continuous optimization of intracellular signaling domains to improve the anti-tumor efficacy and persistence of CAR-T cells, which are fully covered by our plasmid vector products.
1) 1st Generation: Contains only the CD3ζ intracellular signaling domain. It can activate T cell cytotoxicity but has poor proliferation ability and short persistence in vivo, with limited anti-tumor efficacy. Our 1st generation CD274 (PDL1) CAR expression plasmid vector is suitable for basic research on CAR-T cell activation mechanisms.
2) 2nd Generation: Adds one costimulatory domain (e.g., CD28, 4-1BB/CD137) to the CD3ζ domain. This design significantly enhances T cell proliferation, cytokine secretion, and in vivo persistence, improving anti-tumor activity compared to the first generation. It is the most widely used generation in preclinical research.
3) 3rd Generation: Incorporates two costimulatory domains (e.g., CD28+4-1BB, CD28+OX40) along with the CD3ζ domain. It further strengthens T cell activation, expansion, and long-term survival, showing better anti-tumor efficacy in preclinical studies.
4) 4th Generation (Armored CAR): Based on the second generation, it is modified to constitutively or inducibly express cytokines (e.g., IL-12, IL-15) or immune modulators, which can reshape the tumor microenvironment, recruit other immune cells, and enhance the anti-tumor effect of CAR-T cells.
5) 5th Generation (Universal CAR): Optimized for allogeneic CAR-T cell therapy, with modifications such as knockout of PD-1 or CD52 to reduce graft-versus-host disease (GVHD) and improve the safety and efficacy of allogeneic CAR-T cells. It also supports multi-target recognition (e.g., CD274 combined with CLDN18.2) for broader anti-tumor coverage.

Current Research Achievements of CD274 (PDL1) CAR

Approved Drugs Related to CD274 (PDL1)

Research Hotspots of CD274 (PDL1) CAR

Research Difficulties & Challenges of CD274 (PDL1) CAR

Frequently Asked Questions (FAQs)

Q: What is the difference between CD274 (PDL1) CAR-T therapy and PD-1/PD-L1 antibody therapy?
A: PD-1/PD-L1 antibody therapy blocks the interaction between PD-1 and PD-L1 by binding to PD-1 or PD-L1, thereby releasing the inhibitory signal of T cells. CD274 (PDL1) CAR-T therapy directly modifies T cells to express CD274 (PDL1) CAR, enabling T cells to specifically recognize and kill CD274 (PDL1)-positive tumor cells. Compared with antibody therapy, CD274 (PDL1) CAR-T therapy has stronger specificity and longer persistence, and can overcome the drug resistance of antibody therapy.

Q: Which generation of CD274 (PDL1) CAR is suitable for my research?
A: The choice of CAR generation depends on your research goals. If you are conducting basic research on CAR activation mechanism, the 1st generation is suitable. If you are conducting preclinical research on anti-tumor efficacy, the 2nd or 3rd generation is more suitable. If you are exploring combination therapy or allogeneic CAR-T therapy, the 4th or 5th generation is recommended.

Q: What are the main factors affecting the anti-tumor efficacy of CD274 (PDL1) CAR-T cells?
A: The main factors include the specificity and affinity of scFv, the design of signaling domains, the transfection efficiency of plasmid vectors, the persistence of CAR-T cells in vivo, and the tumor microenvironment.

Q: Is CD274 (PDL1) CAR-T therapy safe?
A: CD274 (PDL1) CAR-T therapy has manageable safety profiles in early clinical trials. The main adverse events include cytokine release syndrome (CRS) and neurotoxicity, which can be controlled by appropriate treatment. The 4th and 5th generation plasmid vectors are designed to reduce adverse events by optimizing CAR structure and expressing regulatory molecules.

Q: How should I store the CD274 (PDL1) CAR expression plasmid vector?
A: The plasmid vector should be stored at -20℃ in a refrigerator, avoiding repeated freezing and thawing. When stored properly, the plasmid can remain stable for at least 12 months. For short-term storage (within 3 months), it can be stored at 4℃.

Q: What transfection method is suitable for your plasmid vectors?
A: Our plasmid vectors are compatible with various transfection methods, including liposome-mediated transfection, electroporation, and viral production/transduction. For T cell transfection, electroporation is recommended for higher efficiency.

Q: Can your plasmid vectors be used for in vivo experiments?
A: Yes, our plasmid vectors with viral-based backbones (especially AAV vectors and lentiviral vectors) are suitable for in vivo experiments. AAV vectors have high safety and low immunogenicity, making them ideal for in vivo gene delivery. We can provide guidance on in vivo application of plasmid vectors.

Q: How to verify the expression of CD274 (PDL1) CAR after transfection?
A: You can verify the expression of CAR by flow cytometry (using fluorescent markers on the plasmid, or fluorescently labeled CD274/PDL1 protein, or anti-4GS-linker antibody), Western blot (detecting CAR protein), or qRT-PCR (detecting CAR mRNA). Some of our plasmid vectors are equipped with fluorescent markers, which can be easily detected by flow cytometry.

Q: What should I do if the transfection efficiency is low?
A: Low transfection efficiency may be caused by inappropriate transfection method, cell state, or plasmid quality. You can optimize the transfection protocol (e.g., adjust the ratio of plasmid to transfection reagent, transfection time), improve the cell state.

Q: What information do I need to provide for customized plasmid construction?
A: You need to provide the desired CAR structure (scFv sequence, hinge, transmembrane domain, signaling domain), vector backbone type, markers (fluorescent and antibiotic), and any additional functional elements (e.g., cytokines, gene editing elements). Our technical team will communicate with you to confirm the details.

Q: How long does it take to complete the customized plasmid construction?
A: Generally, the customized plasmid construction can be completed within 7-14 working days. The specific time depends on the complexity of the plasmid design. For urgent needs, we can provide expedited services.

Q: Can you guarantee the functionality of the customized plasmid?
A: Yes, we will conduct strict quality control on the customized plasmid, to ensure that the plasmid meets your research needs. If the plasmid does not meet the requirements, we will re-construct it for free.

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