CD1A Chimeric Antigen Receptor (CAR): A Comprehensive Guide and Our Service & Product Introduction

Our CD1A CAR Expression Plasmid Vector Products and Custom Services

RGBiotech provides a full range of CD1A CAR expression plasmid vectors, covering all generations of CD1A CAR (1st to 5th generation), and supports customized construction services to meet the diverse needs of researchers and enterprises. Our products are widely used in preclinical research and large scale production, with reliable quality and high performance, helping to accelerate the research and development process of CD1A CAR-related therapies.

Product Features

1. Multiple generations available: We provide 1st to 5th generation CD1A CAR expression plasmid vectors, each with unique structural characteristics and functional advantages.
1) 1st generation: Contains only the CD3ζ intracellular signaling domain, suitable for basic research on T-cell activation and CD1A recognition.
2) 2nd generation: Adds one co-stimulatory domain (CD28 or 4-1BB) based on the 1st generation, enhancing T-cell proliferation and persistence, ideal for preclinical efficacy studies.
3) 3rd generation: Contains two co-stimulatory domains (e.g., CD28+4-1BB or CD28+OX40), further improving the anti-tumor activity and in vivo persistence of CD1A CAR-T cells.
4) 4th generation: Modified from the 2nd generation, with inducible or constitutive expression of cytokines, enhancing the anti-tumor effect and reducing adverse reactions.
5) 5th generation: Integrates the intracellular domain of cytokine receptors based on the 2nd generation, further optimizing T-cell function and anti-tumor efficacy for CD1A-positive targets.

2. Diverse vector backbones: We offer a variety of vector backbones to meet different application scenarios, including non-viral vectors (plasmid vectors), lentiviral vectors, retroviral vectors, and AAV (Adeno-Associated Virus) vectors. Lentiviral vectors are suitable for efficient transduction of T cells, retroviral vectors for stable integration and long-term expression, AAV vectors for high safety and low immunogenicity, and non-viral vectors for transient expression and large-scale production.

3. Flexible promoter options: Our vectors are equipped with a variety of high-efficiency promoters to meet different expression needs, including CMV promoter (strong constitutive expression), EF1α promoter (stable expression in mammalian cells) and APC/T-cell-specific promoters (for targeted expression in antigen-presenting cells or T cells), ensuring high and stable expression of CD1A CAR.

4. Multiple fluorescent and antibiotic selection markers: Fluorescent markers include GFP (green fluorescent protein), RFP (red fluorescent protein), luciferase etc., which facilitate the observation and sorting of transfected cells. Antibiotic selection markers include Puromycin (Pur), Neomycin (Neo), Hygromycin (Hygro) and Blasticidin (Bla), enabling efficient screening of positive clones. We also provide vectors with dual markers for more flexible experimental design.

Product Advantages

Product Applications

Customized Plasmid Vector Construction Services

Introduction of CD1A

CD1A (CD1a Molecule) is a protein encoded by the CD1A gene in humans (Gene ID: 909, Ensembl: ENSG00000158477). The human CD1A gene is located on chromosome 1q23.1, spanning from 158248336 bp to 158258269 bp on the GRCh38.p14 assembly, and is part of a cluster of five CD1 family genes organized on chromosome 1. It is also known by synonyms such as R4, T6, CD1, FCB6, and HTA1, with a UniProt ID of P06126. The CD1A gene is highly conserved in mammals and plays a key role in lipid antigen presentation and adaptive immune response, with its abnormal expression closely associated with various T-cell malignancies and inflammatory diseases.

CD1A is a member of the CD1 family of transmembrane glycoproteins, structurally related to major histocompatibility complex (MHC) proteins, and forms heterodimers with beta-2-microglobulin (β2m). It is an asparagine-glycosylated protein with a molecular weight of approximately 49 kDa (33 kDa polypeptide backbone). Its structure consists of an extracellular immunoglobulin-like domain (with a small antigen-binding groove and stunted a′ pocket), a single transmembrane domain, and a short cytoplasmic tail. The extracellular domain is responsible for binding lipid and glycolipid antigens, while the transmembrane domain anchors the protein to the cell membrane. The cytoplasmic tail lacks intrinsic kinase activity but participates in intracellular trafficking, with the protein localizing to the plasma membrane and recycling vesicles of the early endocytic system.

CD1A primarily mediates the presentation of lipid and glycolipid antigens (of self or microbial origin) to T cells, playing a crucial role in adaptive immune responses and lipid-reactive T cell activation. It is involved in regulating T cell differentiation and function, particularly in the activation of CD1A-restricted T cells that produce Th2 or Th17 cytokines. These T cells contribute to immune responses against pathogens and are also implicated in inflammatory processes. Additionally, CD1A is critical for the development and function of antigen-presenting cells (APCs), including Langerhans cells, and participates in the recognition of foreign lipids to initiate immune defense.

CD1A is predominantly expressed on professional antigen-presenting cells and thymocytes, with a biased expression pattern in human tissues. It is highly and exclusively expressed on Langerhans cells (a subset of dendritic cells in the skin) and double-positive (CD4+CD8+) cortical thymocytes, with lower expression on single-positive (CD4+ or CD8+) thymocytes. In peripheral tissues, CD1A is mainly expressed in the skin (including thigh, hip, arm, and abdomen skin), esophagus, thymus, testicle, gums, and monocytes. Notably, CD1A is not expressed on mature T cells, B cells, NK cells, or CD34+ hematopoietic stem cells, making it a safe and selective target for immunotherapy of T-cell malignancies.

CD1A is closely associated with T-cell malignancies, inflammatory skin diseases, and pulmonary disorders. In T-cell malignancies, it is specifically expressed on cortical T-cell acute lymphoblastic leukemia (cT-ALL) cells, a subtype accounting for approximately 30% of T-ALL cases. CD1A expression is rare in relapsed/refractory (r/r) T-ALL, with only 8% of cases showing high expression (≥80%), but it remains a valuable target for select patients with cT-ALL. In inflammatory skin diseases, CD1A-restricted T cells contribute to the pathogenesis of atopic dermatitis, psoriasis, and allergic contact dermatitis. Additionally, CD1A transcript levels are upregulated in the lung parenchyma of smokers, suggesting a potential role in pulmonary disorders. CD1A’s restricted expression on malignant cells and absence on normal mature T cells makes it an ideal target to minimize off-tumor toxicity.

Introduction of CD1A Chimeric Antigen Receptor (CAR)

CD1A Chimeric Antigen Receptor (CD1A CAR) is a genetically engineered receptor designed to specifically recognize the CD1A antigen on the surface of target cells. Its structure typically includes four main components: an extracellular antigen-recognition domain (single-chain variable fragment, scFv) that binds to CD1A, an extracellular hinge region (e.g., CD8α stalk) that provides flexibility and stability, a transmembrane domain that anchors the receptor to the T-cell membrane, and an intracellular signaling domain that mediates T-cell activation. CD1A CAR-modified T cells (CD1A CAR-T cells) can specifically recognize and kill CD1A-positive tumor cells, with the advantage of being fratricide-resistant due to the absence of CD1A on normal mature T cells.

Recent research on CD1A CAR has focused on its application in the treatment of cT-ALL, with promising preclinical and early clinical results. Preclinical studies have demonstrated that CD1A CAR-T cells exhibit potent anti-tumor activity against CD1A-positive T-ALL cell lines and primary cT-ALL samples, with no significant off-tumor toxicity. In patient-derived xenograft (PDX) models, CD1A CAR-T cells showed excellent in vivo persistence and significant tumor-killing efficacy. A clinical stage biotech company (One Chain) is developing an autologous CD1A CAR-T therapy (OC-1) for r/r cT-ALL, which has received Orphan Drug Designation (ODD) from the FDA and EMA. The first-in-human (FIH) phase 1/2 clinical trial is ongoing (dose level 2 currently recruiting), with a validated GMP manufacturing process and completed non-clinical package demonstrating safety and efficacy.
Additional preclinical studies have confirmed that CD1A CAR-T cells are fratricide-resistant, as CD1A is not expressed on normal mature T cells, circumventing a major challenge in T-cell malignancy CAR-T therapy. Retrospective studies on r/r cT-ALL patients have been completed, with results under review, and long-term follow-up studies are ongoing to monitor the safety and persistence of CD1A CAR-T therapy.

Currently, there are no CD1A CAR-T cell drugs officially approved by the U.S. Food and Drug Administration (FDA), European Medicines Agency (EMA), or National Medical Products Administration (NMPA) globally. However, multiple CD1A CAR-T products are in early clinical trial stages (phase 1/2), with OC-1 (One Chain) being the most advanced candidate, focusing on the treatment of r/r cT-ALL. Other preclinical CD1A CAR-T programs are exploring applications in other CD1A-positive diseases, including inflammatory skin disorders. With the continuous advancement of clinical research, it is expected that the first CD1A CAR-T drug will be approved for marketing in the next 3-5 years, addressing the unmet medical need for r/r cT-ALL patients.

CD1A CAR Research Hotspots

CD1A CAR Research Difficulties & Challenges

1. Low CD1A expression in r/r T-ALL: CD1A is highly expressed in only 8% of r/r T-ALL cases, with most refractory or relapsed patients showing CD1A negativity. This poses significant recruitment challenges for clinical trials and limits the patient population that can benefit from CD1A CAR-T therapy.
2. Tumor heterogeneity and antigen loss: CD1A expression can vary within the same tumor, leading to tumor heterogeneity. Some malignant cells may downregulate or lose CD1A expression, resulting in immune escape and treatment relapse, which is a major challenge affecting long-term efficacy.
3. Limited clinical data: Most CD1A CAR-T studies are in early clinical stages (phase 1/2) with small patient cohorts. Limited long-term safety and efficacy data make it difficult to fully evaluate the clinical value of CD1A CAR-T therapy, especially in terms of long-term survival and late adverse reactions.
4. Manufacturing complexity: Despite being fratricide-resistant, the preparation of CD1A CAR-T cells still requires complex technologies such as gene editing, cell culture, and quality control, resulting in high manufacturing costs that limit its accessibility.
5. Off-tumor toxicity concerns: While CD1A is not expressed on mature T cells, it is expressed on Langerhans cells and cortical thymocytes. Potential off-tumor toxicity to these normal cells remains a concern, requiring further optimization of CD1A CAR specificity to minimize tissue damage.

Frequently Asked Questions (FAQs)

Q: What is the difference between different generations of CD1A CAR expression plasmid vectors?
A: The main difference lies in the composition of the intracellular signaling domain. The 1st generation only contains CD3ζ; the 2nd generation adds one co-stimulatory domain (CD28 or 4-1BB), which enhances T-cell persistence; the 3rd generation contains two co-stimulatory domains; the 4th generation can express cytokines; the 5th generation integrates the intracellular domain of cytokine receptors. With the increase of generations, the anti-tumor activity, proliferation ability, and in vivo persistence of CD1A CAR-T cells are gradually improved.

Q: Which vector backbone should I choose for CD1A CAR-T cell preparation?
A: It depends on your experimental needs. Lentiviral vectors are suitable for efficient transduction of T cells and stable long-term expression; retroviral vectors are suitable for dividing cells and stable integration; AAV vectors have high safety and low immunogenicity, suitable for in vivo delivery; non-viral plasmid vectors are suitable for transient expression, large-scale production, and low-cost experiments.

Q: How to choose the appropriate promoter for CD1A CAR expression plasmid vectors?
A: CMV promoter is suitable for strong constitutive expression in most mammalian cells; EF1α promoter has stable expression and is not easily silenced, suitable for long-term cell culture; APC/T-cell-specific promoters are suitable for targeted expression in antigen-presenting cells or T cells. If you are not sure, we recommend choosing CMV or EF1α promoter for general research.

Q: What are the advantages of fluorescent markers in CD1A CAR expression plasmid vectors?
A: Fluorescent markers (such as GFP, RFP) can help you quickly observe the transfection efficiency of the vector, sort transfected positive cells by flow cytometry, and track the expression and distribution of CD1A CAR in cells and animal models, which is very important for verifying the success of vector delivery and CAR expression.

Q: What is the delivery time for customized plasmid vector construction services?
A: The delivery time depends on the complexity of the customized project. Generally, the standard customized project can be delivered within 2-4 weeks, and the complex project (such as multi-functional element addition, large-scale preparation) can be delivered within 4-8 weeks. We will confirm the delivery time with you before starting the project and ensure on-time delivery.

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