Recombinant Human CD40 ligand (CD40LG), partial (Active)

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Description

Molecular Composition

  • Expression Region: Covers amino acids 113–261 (C194W mutation) .

  • Molecular Weight: ~16–17 kDa (monomer) .

  • Tag-Free Design: Ensures minimal interference with biological activity .

  • Purity: >95% as confirmed by SDS-PAGE and HPLC .

  • Endotoxin Level: <1.0 EU/µg (LAL method) .

ParameterValueSource
Expression SystemE. coli
FormulationLyophilized powder
Storage Stability-20°C or -80°C for long-term use
ReconstitutionSterile water or PBS (0.1 mg/mL)

Functional Role in Immune Signaling

CD40LG binds to CD40 receptors on APCs (e.g., dendritic cells, B cells), triggering downstream pathways:

  • Canonical NF-κB Pathway: Drives pro-inflammatory cytokine production (e.g., IL-6, TNF-α) .

  • Non-Canonical NF-κB Pathway: Mediates long-term immune responses via NIK accumulation .

  • MAPK and Akt Pathways: Activates JNK, p38, and PI3K/Akt for cell survival and proliferation .

ED50 Values:

  • IL-8 Induction: <5–10 ng/mL in human PBMCs .

  • HEK-Blue CD40L Assay: ≤50 ng/mL .

Key Experimental Uses

ApplicationDetailsReferences
Cancer ImmunologyEnhances T cell–APC interactions to boost anti-tumor responses .
Autoimmune DiseasesModels immune dysregulation in conditions like lupus or rheumatoid arthritis.
Vaccine DevelopmentPotentiates antibody class switching and memory B cell formation .
Gene TherapyReplaces defective CD40LG in X-linked hyper IgM syndrome (XHM) .

Case Study: X-Linked Hyper IgM Syndrome (XHM)

In a clinical trial, recombinant CD40L (rCD40L) was administered subcutaneously to XHM patients:

  • Dosage: 0.03 mg/kg (22 weeks) → 0.05 mg/kg (additional 22 weeks) .

  • Outcomes:

    • T Cell Function: Restored IFN-γ and TNF-α production .

    • Delayed Hypersensitivity: Induced cutaneous reactions (reversed post-treatment) .

    • Limitations: No improvement in T cell–dependent antibody responses .

  • Safety: Mild liver enzyme elevations at higher doses (0.05 mg/kg) .

Comparative Analysis of Recombinant CD40LG Products

Catalog NumberExpression RegionMW (kDa)PurityEndotoxinED50 (IL-8)Source
CSB-AP002211HU113–261aa (C194W)16.2>95%<1.0 EU/µg<5–10 ng/mL
RPC29236113–261aa16.2>95%<1.0 EU/µgN/A
RPC28868113–261aa (C194W)16.3>95%<1.0 EU/µgN/A
Z100125Full-length soluble17>95%<1.0 EU/µg<5 ng/mL
orb594873113–261aa16.2>95%<1.0 EU/µg≤10 ng/mL

Key Signaling Complexes

  1. TRAF Recruitment:

    • TRAF2/3: Mediates NF-κB and JNK activation .

    • cIAP1/2: Regulates NIK stability in non-canonical NF-κB signaling .

  2. Oligomerization:

    • Trimerization of CD40LG enhances receptor clustering and signaling efficacy .

Technical Considerations

  • Stability: Lyophilized form requires careful reconstitution to avoid aggregation .

  • Dosing: Optimal therapeutic levels balance efficacy and hepatotoxicity .

Emerging Applications

  • Cancer Immunotherapy: Synergies with checkpoint inhibitors to enhance T cell infiltration .

  • Gene Editing: CRISPR/Cas9-mediated correction of CD40LG mutations in XHM .

Product Specs

Buffer
Lyophilized from a 0.2 µm filtered PBS, pH 7.0
Description

Recombinant Human CD40 ligand (CD40LG) is a valuable tool for cancer research, particularly in the field of immunology. CD40LG, also known as CD40 ligand or T-cell antigen Gp39, is a member of the tumor necrosis factor (TNF) ligand superfamily. It plays a crucial role in immune response regulation and activation of various immune cells.

This high-quality recombinant protein is derived from an E. coli expression system and encompasses amino acids 113 to 261 (C194W). Presented in a tag-free format, it ensures purity and minimizes potential interference from extraneous tags. With a purity exceeding 95%, as determined by SDS-PAGE and HPLC, this protein offers consistency and reliability for your experiments.

Our CD40LG exhibits full biological activity, comparable to a standard reference. It possesses an effective dose (ED50) of less than 5-10 ng/ml in the dose-dependent stimulation of interleukin-8 (IL-8) production by human peripheral blood mononuclear cells (PBMC). The product is supplied as a lyophilized powder, providing convenience and stability for long-term storage and ease of use in research applications.

Form
Lyophilized powder
Lead Time
5-10 business days
Notes
Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Reconstitution
We recommend centrifuging the vial briefly prior to opening to ensure the contents settle to the bottom. Reconstitute the protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL. We suggest adding 5-50% glycerol (final concentration) and aliquoting for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers can use this as a reference.
Shelf Life
The shelf life is influenced by various factors, including storage conditions, buffer ingredients, storage temperature, and the inherent stability of the protein itself.
Generally, the shelf life of the liquid form is 6 months at -20°C/-80°C. The shelf life of the lyophilized form is 12 months at -20°C/-80°C.
Storage Condition
Upon receipt, store at -20°C/-80°C. Aliquoting is necessary for multiple uses. Avoid repeated freeze-thaw cycles.
Tag Info
Tag-Free
Synonyms
CD 40L; CD154; CD40 antigen ligand; CD40 ligand; CD40 ligand; soluble form; CD40-L; CD40L; CD40L_HUMAN; CD40LG; gp39; hCD40L; HIGM1; IGM; IMD3; T B cell activating molecule; T BAM; T-cell antigen Gp39; TNF-related activation protein; TNFSF5; TrAP; Tumor necrosis factor (ligand) superfamily member 5; Tumor necrosis factor ligand superfamily member 5
Datasheet & Coa
Please contact us to get it.
Expression Region
113-261aa(C194W)
Mol. Weight
16.3 kDa
Protein Length
Partial
Purity
>95% as determined by SDS-PAGE.
Research Area
Cancer
Source
E.coli
Species
Homo sapiens (Human)
Target Names
Uniprot No.

Target Background

Function

CD40 ligand is a cytokine that functions as a ligand to CD40/TNFRSF5. It acts as a costimulator for T-cell proliferation and cytokine production. Its cross-linking on T-cells generates a costimulatory signal that enhances the production of IL4 and IL10 in conjunction with TCR/CD3 ligation and CD28 costimulation. CD40 ligand induces the activation of NF-kappa-B, kinases MAPK8 and PAK2 in T-cells, and tyrosine phosphorylation of isoform 3 of CD28. In the absence of co-stimulus, it mediates B-cell proliferation and in the presence of IL4, it induces IgE production. It is involved in immunoglobulin class switching.

CD40 ligand acts as a ligand for integrins, specifically ITGA5:ITGB1 and ITGAV:ITGB3. Both integrins and the CD40 receptor are required for the activation of CD40-CD40LG signaling, which exerts cell-type dependent effects, such as B-cell activation, NF-kappa-B signaling, and anti-apoptotic signaling.

Gene References Into Functions
  1. This study reports the clinical and CD40L genetic features of six Iranian hyper IgM syndrome patients. PMID: 30081731
  2. Overexpression of CD40L-WT/CD40L-M in CD40+ NSCLC cells increased SA-beta-gal staining activity and inhibited DNA synthesis and cell proliferation. PMID: 30078020
  3. T cell-stimulated CLL cells actively recruited monocytes, and CD40L was identified as the responsible T-cell factor that mediated recruitment. PMID: 28971904
  4. This study shows that decreased serum levels of soluble CD40L correlate in treated patients with Relapsing-Remitting Multiple Sclerosis. PMID: 29050818
  5. In conclusion, we described a new pathway of platelet-monocyte interaction, mediated by sCD40L and oxidative stress that may contribute to the progression of endothelial dysfunction during Shiga toxin 2-associated hemolytic uremic syndrome. PMID: 29068360
  6. This study shows that sCD40L/alpha5beta1 interaction leads to platelet activation as evaluated in the human whole blood. PMID: 26719354
  7. In this first X chromosome-wide association study of adult patients with IBD, we identified an IBD susceptibility locus with genome-wide significance at rs2427870 on chrXq26.3, located 66 kbp upstream of CD40LG and 83.4 kbp upstream of ARHGEF6 [OR, 1.22; combined p = 3.79 x 10-15]. PMID: 28333213
  8. Study indicates that the rs1126535C/T polymorphism of CD154 gene was involved in the progression of Chinese SLE patients, probably by affecting the expression of CD154. PMID: 28550400
  9. This study found that plasma CD40L was associated with acute chest syndrome (ACS), and that sickle cell anemia (SCA)patients with a lifetime history of ACS (ACS+) presented significantly higher plasma CD40L and TSP-1 than patients who had never experienced ACS (ACS-). PMID: 28609750
  10. This paper demonstrates the importance of CD40/CD40L signaling on IL-10-producing regulatory B cells in Chinese children with Henoch-Schonlein purpura nephritis. PMID: 27837410
  11. Soluble CD40 ligand derived from serum is not correlated with the early stage of multiple sclerosis. PMID: 28619427
  12. Analysis of the early events after receptor engagement revealed that both TNF and CD40L activate the classical NF-kappaB pathway, and confirm activation of the alternative by the latter. Furthermore, using genetic and pharmacological inhibition of the classical pathway we show that activation of the alternative occurs independently of the former. This reveals insights into NF-kappaB signaling by CD40L and TNF in endothelial cells. PMID: 29183724
  13. CD40L, more than IL-6, or TNF-alpha, constitutes a predictor to explain polycystic ovary syndrome and associated features. PMID: 27572328
  14. Soluble CD40 ligand directly alters glomerular permeability and may act as a circulating permeability factor in focal segmental glomerulosclerosis. PMID: 29155846
  15. Plasma sCD40L levels were elevated in systemic lupus erythematosis patients who had positive anti-phospholipid antibodies and experienced arterial thrombosis, suggesting that enhanced release of sCD40L through platelet activation presumably by aPL could contribute to the development of atherothrombotic disease. PMID: 28421990
  16. Higher concentrations of CD40L in patients with limited cutaneous form in our study might suggest a role for the CD40/CD40L pathway in vascular pathology of systemic sclerosis. PMID: 27392528
  17. We also report for the first time that the rs1126535 C allele (CD40L gene) may predict a worse response after gastric bypass in morbidly obese patients. PMID: 27681093
  18. Serum levels of sCD40L and MMP-9 are associated with the stability of carotid plaques. PMID: 28642174
  19. Studies identify a novel molecular mechanism of regulation of CD40L by the transcription factor GLI2 in the tumor microenvironment downstream of CCR3 signaling. PMID: 28461568
  20. Data suggest that therapeutic CD40-CD40L blocking agents may prove efficacious not only in early and established rheumatoid arthritis (RA), but also in inhibiting the progression of the disease from arthralgia or undifferentiated arthritis to RA. PMID: 28455435
  21. CD4(+) T cells that coexpress CD57 and CD154, which are exclusively present in cytomegalovirus-positive individuals. PMID: 27566833
  22. Serum CD40L levels were elevated in both neuromyelitis optica and multiple sclerosis patients. PMID: 27725124
  23. Plasma PGE2 is correlated with the prevention of IVIG resistance and CAL formation through CD40L in KD. PMID: 27525421
  24. This study provides the first evidence that human circulating group 2 innate lymphoid cells can express CD154 and stimulate the production of IgE by B lymphocytes through IL-25/IL-33 stimulation or TLR triggering. PMID: 27576126
  25. Data also demonstrated that the CD154-triggered inhibition of the Fas-mediated cell death response was dependent on a suppression of caspase-8 cleavage, but independent of de novo protein synthesis or alterations in Fas expression on the cell surface. PMID: 27391025
  26. These results suggest soluble CD40L could have a prognostic value in ST-elevation myocardial infarction patients. PMID: 27172386
  27. Persistence of helper T-cell-derived CD40L on or in B cells could permit sustained CD40 signaling enabling survival and proliferation of antigen-presenting B cells following brief interactions with helper T cells in vivo in germinal centers. PMID: 27753080
  28. These results demonstrate the feasibility of engineered nuclease-directed gene repair to restore endogenously regulated CD40L, and the potential for its use in T-cell therapy for X-HIGM syndrome. PMID: 26903548
  29. Hypertensives showed significantly enhanced soluble CD40L levels compared to normotensive controls. PMID: 27090943
  30. The levels of sCD40l have no influence on survival or cardiovascular events and mortality in haemodialysis patients in a long-term follow-up. PMID: 27295448
  31. CD40L gene polymorphism was found to be associated with severe falciparum malaria in the Indian population, especially in severe malarial anemia. PMID: 28352049
  32. While CD40 expression tends to be relatively high in the peritumoral dermis of epithelial carcinomas, the expression of CD40L in mast cells is low in the same peritumoral area, compared with the opposite findings in psoriasis and actinic keratosis. PMID: 28267402
  33. Increased serum sCD40L levels may be related to angiogenesis in patients with multiple myeloma (MM). This protein has potential clinical usefulness in MM and may be considered as an additional prognostic marker. The correlation of sCD40L with beta-thromboglobulin may indicate that in patients with MM sCD40L derives from activated platelets. PMID: 27243341
  34. Primary Aldosteronism is related to platelet activation, expressed as higher plasma values of soluble CD40L and soluble P-selectin values. PMID: 27101095
  35. Given the crucial role of sCD40L, this haplotype study in a transfusion model may be helpful to further determine the role of haplotypes in inflammatory clinical settings. PMID: 27094978
  36. Overexpression of CD154 on CD4(+)T cells is unlikely to be central to the pathogenesis of idopathic thrombocytopenic purpura, and other immune dysfunctions should be targeted for therapy purposes. PMID: 26183367
  37. The serum levels of the soluble factors sCD40L and CXCL1 are not associated with endometriosis and are not suitable as biomarkers for disease diagnosis. PMID: 27190986
  38. Increased sCD40L plasma levels are associated with the presence of insulin resistance and not the state of glucose tolerance. PMID: 26934129
  39. We report that Kv1.3-NPs reduced NFAT activation and CD40L expression exclusively in CD45RO(+) T cells. Furthermore, Kv1.3-NPs suppressed cytokine release and induced a phenotype switch of T cells from predominantly memory to naive. PMID: 26994905
  40. mCD40L-induced cell death mediated by NORE1A expression appeared to be independent of mCD40L-induced cell death mediated by sustained JNK activation since NORE1A inhibition did not affect JNK phosphorylation and vice versa. PMID: 26986513
  41. The diagnostic value of soluble CD40 ligand (sCD40L) and vascular endothelial growth factor (VEGF) for Alzheimer's disease. PMID: 26706786
  42. Study allows identification of significant different genetic heterogeneity between two investigated populations (France and Tunisia) and revealed discrepancies in the prevalence of CD40LG polymorphisms that may be explained by ethnic and geographic differences. PMID: 26577033
  43. Concentrations of the C-reactive protein, myeloperoxidase, and soluble CD40 ligand taken from the peripheral vein were closely similar to the concentration found in coronary blood of ACS patients. PMID: 26576922
  44. Studies suggest that the CD40/CD154 pathway represents a promising potential therapeutic target for the prevention of transplantation rejection. PMID: 26268734
  45. Three single-nucleotide polymorphisms (SNPs) of the TLR8, CD40LG, and IRAK1 genes on the X chromosome were genotyped. PMID: 26043172
  46. Plasma soluble CD40L levels are reduced by antiplatelet therapy with clopidogrel, but not associated with long-term ischemic outcomes in unselected consecutive aspirin-treated patients undergoing cardiac catheterization. PMID: 26237513
  47. The study findings showed that plasma sCD40L, fetuin-A, and PAPP-A levels are associated with carotid plaque formation and instability. PMID: 26214492
  48. CD40 ligand induces RIP1-dependent, necroptosis-like cell death in low-grade serous but not serous borderline ovarian tumor cells. PMID: 26313915
  49. CD40 ligand induces von Willebrand factor release from endothelial cells. PMID: 25608503
  50. Higher plasma soluble CD40L levels on presentation are associated with clinical severity and have the potential to be a good prognostic biomarker of aneurysmal subarachnoid hemorrhage. PMID: 25944664

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Database Links

HGNC: 11935

OMIM: 300386

KEGG: hsa:959

STRING: 9606.ENSP00000359663

UniGene: Hs.592244

Involvement In Disease
Immunodeficiency with hyper-IgM, type 1 (HIGM1)
Protein Families
Tumor necrosis factor family
Subcellular Location
Cell membrane; Single-pass type II membrane protein. Cell surface.; [CD40 ligand, soluble form]: Secreted.
Tissue Specificity
Specifically expressed on activated CD4+ T-lymphocytes.

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