TNFSF7 Human

What is TNFSF7 and what are its known synonyms in scientific literature?

TNFSF7 is a cytokine that belongs to the tumor necrosis factor ligand superfamily. It is commonly known as CD70, but also appears in scientific literature under multiple synonyms including CD27L (CD27 Ligand), CD27LG, CD27-L, Surface Antigen CD70, and Ki-24 Antigen . When searching literature databases, researchers should include these alternative designations to ensure comprehensive results.

What is the molecular structure and basic properties of human TNFSF7?

Human TNFSF7 (CD70) is a 30 kDa type 2 transmembrane glycoprotein containing three distinct regions: a 20 amino acid cytoplasmic region, a transmembrane domain, and a 155 amino acid extracellular domain characterized by multiple beta-strands . The recombinant form produced in E. coli is a single, non-glycosylated polypeptide chain containing 178 amino acids (spanning positions 39-193) with a molecular mass of approximately 19.5 kDa . The protein functions as an inducible trimer that appears on the surface of activated B cells, T cells, and NK cells .

What are the primary physiological functions of TNFSF7?

TNFSF7 plays several critical roles in immune function:

• Participation in T-cell activation mechanisms

• Induction of proliferation in costimulated T-cells

• Enhancement of cytolytic T-cell generation

• Regulation of T-cell dependent B-cell differentiation into plasma cells

• Promotion of clonal expansion of T cells

These functions make TNFSF7 an important mediator in adaptive immune responses and potentially significant in immunotherapy research.

What pathological conditions have been associated with TNFSF7?

Research has identified associations between TNFSF7 and several pathological conditions:

• Acute myocarditis and myocarditis

• Possible involvement in migraine without aura (MO), as suggested by SNP association analysis

• While not explicitly mentioned in the provided sources, other research has implicated TNFSF7 in autoimmune disorders and certain malignancies

What are the optimal storage and stability conditions for working with recombinant TNFSF7?

Researchers should follow these evidence-based protocols for optimal stability:

For short-term use (2-4 weeks):

• Store at 4°C in the provided formulation

For longer-term storage:

• Store frozen at -20°C to -70°C

• Add a carrier protein (0.1% HSA or BSA) for extended storage periods

• Avoid multiple freeze-thaw cycles which can degrade protein integrity

The recommended formulation for TNFSF7 protein solution is 1mg/ml containing 20mM Tris-HCl buffer (pH 8.0) and 10% glycerol .

What methods are available for detecting TNFSF7 in experimental samples?

Several validated methods can be employed for TNFSF7 detection:

• Western Blot Analysis:

  • Successfully demonstrated using THP-1 human acute monocytic leukemia cell line lysates

  • PVDF membrane probing with specific anti-human CD27 Ligand/TNFSF7 antibodies (e.g., Biotinylated Antigen Affinity-purified Polyclonal Antibody) followed by HRP-conjugated secondary antibodies

  • Specific band detection at approximately 33-35 kDa under reducing conditions

• Flow Cytometry:

  • Utilizing fluorophore-conjugated antibodies (e.g., Alexa Fluor 700-conjugated anti-TNFSF7)

  • Particularly useful for detecting expression on specific immune cell populations

• SDS-PAGE:

  • For purity assessment, with recombinant TNFSF7 showing greater than 80% purity

How should optimal antibody dilutions be determined for TNFSF7 detection?

When working with antibodies against TNFSF7:

• Optimal dilutions must be determined empirically by each laboratory for specific applications

• Start with manufacturer-recommended concentrations (e.g., 0.1 μg/mL for Western blot applications as demonstrated with THP-1 cell lysates)

• Perform titration experiments across a range of concentrations

• Include appropriate positive controls (such as activated T cells or B cells known to express TNFSF7)

• Validate specificity using negative controls (resting lymphocytes typically show minimal TNFSF7 expression)

What are the interspecies differences in TNFSF7 that researchers should consider in translational studies?

Researchers conducting translational studies or using animal models should consider these comparative aspects:

• Human CD27 Ligand/TNFSF7 extracellular region shares only 64% amino acid identity with mouse and rat homologs

• This moderate homology suggests potential functional differences across species

• When interpreting animal model data, these differences may impact:

  • Binding affinity to receptors

  • Downstream signaling potency

  • Therapeutic targeting strategies

  • Antibody cross-reactivity

For cross-species studies, validation of reagent specificity and consideration of potential functional differences are essential for accurate data interpretation.

What genetic analysis approaches have been used to study TNFSF7 in disease association studies?

Researchers investigating TNFSF7 genetic associations have employed several methodologies:

• SNP Association Analysis:

  • Used to investigate potential links between TNFSF7 polymorphisms and migraine

  • Case-control study design with matched populations

  • Statistical analysis employing chi-square tests for genotype and allele frequencies

  • Subgroup analysis (e.g., migraine with aura versus migraine without aura)

• Chromosome Location Analysis:

  • TNFSF7 has been mapped to chromosome 19p13

  • Linkage studies have examined this region in relation to migraine susceptibility

• Sequence Analysis:

  • While not explicitly used for TNFSF7 in the provided sources, exon sequencing approaches (as used for the NOTCH3 gene) represent a methodology applicable to TNFSF7 investigations

What are the significant challenges in purifying and maintaining functional TNFSF7 for research applications?

Researchers working with TNFSF7 should address these purification and functionality challenges:

• Protein Stability Issues:

  • TNFSF7 may lose activity during freeze-thaw cycles

  • Carrier proteins (0.1% HSA or BSA) are recommended for long-term storage

  • Temperature sensitivity requires careful handling protocols

• Purification Considerations:

  • Recombinant TNFSF7 is purified using proprietary chromatographic techniques

  • His-tag fusion (23 amino acids at N-terminus) enables affinity purification

  • Purity assessment by SDS-PAGE typically yields >80% purity

• Functional Assessment:

  • Biological activity testing should include T-cell proliferation assays

  • Binding assays to confirm interaction with CD27 receptor

  • Verification of trimeric formation for proper functionality

How can TNFSF7 be effectively used in immunological research protocols?

TNFSF7 can be implemented in immunological research through these methodological approaches:

• T-cell Activation Studies:

  • TNFSF7 can be used to stimulate T-cells in conjunction with other activating signals

  • Measure proliferation, cytokine production, and cytolytic activity

  • Compare effects on different T-cell subpopulations

• B-cell Differentiation Research:

  • Investigate TNFSF7's role in T-cell dependent B-cell differentiation

  • Monitor plasma cell formation and antibody production

  • Examine memory B-cell development

• Co-stimulation Research:

  • Use in combination with other immune modulators to study synergistic effects

  • Investigate signaling pathways activated by TNFSF7-CD27 interaction

  • Explore potential therapeutic applications in immune modulation

What evidence supports the investigation of TNFSF7 in neurological disorders like migraine?

The investigation of TNFSF7 in migraine is supported by several lines of evidence:

• Genetic Association Findings:

  • SNP association analysis revealed a possible association between TNFSF7 and migraine without aura (MO)

  • Statistical significance was observed in genotype analysis (P = 0.036) and allele analysis (P = 0.017) specifically in the MO subgroup

• Chromosomal Location:

  • TNFSF7 is located on chromosome 19p13, a region previously linked to migraine susceptibility

  • This positional relevance strengthens the rationale for investigating TNFSF7 in migraine pathophysiology

• Functional Relevance:

  • TNFSF7 is homologous to TNF-alpha and TNF-beta ligands, which have previously been associated with migraine

  • This homology suggests potential involvement in inflammatory mechanisms relevant to migraine pathogenesis

These findings indicate that further independent studies investigating TNFSF7 in migraine and potentially other neurological disorders are warranted.

What are the methodological considerations when investigating potential associations between TNFSF7 and disease states?

When designing studies to investigate TNFSF7 in disease associations, researchers should consider:

• Study Design Elements:

  • Case-control matching criteria (age, sex, ethnicity)

  • Sample size calculations based on expected effect sizes

  • Appropriate control groups for comparison

  • Subgroup analyses based on clinical phenotypes (as demonstrated in the migraine study separating MO from MA)

• Statistical Analysis Approaches:

  • Chi-square testing for genotype and allele frequencies

  • Correction for multiple testing when examining multiple SNPs

  • Haplotype analysis when multiple markers are available

  • Power calculations to ensure adequate sample sizes

• Validation Requirements:

  • Independent replication in different populations

  • Functional studies to elucidate mechanisms

  • Meta-analysis when multiple studies become available

What technical considerations should researchers address when developing TNFSF7-targeted therapeutic approaches?

Researchers developing TNFSF7-targeted therapeutics should address these technical considerations:

• Target Specificity:

  • TNFSF7 shows 64% homology between human and rodent models, requiring species-specific targeting strategies

  • Confirmation of antibody specificity across different experimental systems

  • Evaluation of potential cross-reactivity with other TNF superfamily members

• Functional Assessment:

  • Verification that targeting approaches affect the known biological functions of TNFSF7

  • Measurement of T-cell activation, proliferation, and cytolytic activity

  • Evaluation of effects on B-cell differentiation

• Delivery System Optimization:

  • Consideration of TNFSF7's expression primarily on activated immune cells

  • Development of strategies to target specific cell populations

  • Evaluation of potential off-target effects on immune function

How might TNFSF7 research contribute to understanding disease mechanisms in cardiac conditions?

TNFSF7 has been associated with cardiac conditions, particularly myocarditis . Research approaches to elucidate its role include:

• Expression Analysis:

  • Quantification of TNFSF7 levels in cardiac tissue from patients with myocarditis compared to controls

  • Correlation of expression levels with disease severity and clinical outcomes

  • Identification of specific cardiac cell populations expressing TNFSF7

• Functional Studies:

  • Investigation of how TNFSF7-CD27 signaling affects cardiac inflammation

  • Assessment of TNFSF7's role in recruitment and activation of immune cells in cardiac tissue

  • Evaluation of potential therapeutic interventions targeting this pathway

• Genetic Association Studies:

  • Examination of TNFSF7 polymorphisms in patients with myocarditis

  • Analysis of potential epistatic interactions with other immune-related genes

  • Longitudinal studies correlating genetic variants with disease progression

This research direction could potentially identify novel therapeutic targets for cardiac inflammatory conditions and improve understanding of disease pathogenesis.