GNLY Antibody
Description
Definition and Function of GNLY Antibody
The Granulysin (GNLY) antibody is a specific immunoglobulin designed to detect and study the Granulysin protein, a member of the saposin-like protein (SAPLIP) family. GNLY is expressed by cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells, functioning as an antimicrobial agent, tumor cell lytic factor, and alarmin that activates immune responses . The antibody enables researchers to analyze GNLY expression, localization, and functional roles in immune processes, infection, and cancer .
Applications of GNLY Antibody
GNLY antibodies are utilized across multiple experimental techniques:
Types of GNLY Antibodies
Multiple antibody formats are available, each optimized for specific experimental needs:
Key Research Findings with GNLY Antibody
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Immune Activation: GNLY activates antigen-presenting cells (APCs) via TLR4, promoting adaptive immune responses . Antibodies confirmed this by blocking GNLY-mediated DC activation .
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Tumor Rejection: Transgenic mice expressing human GNLY exhibited enhanced tumor rejection, validated using GNLY-specific antibodies .
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Infection and Autoimmunity: Elevated GNLY levels correlate with immune defense against Mycobacterium tuberculosis and tissue damage in Stevens-Johnson syndrome .
Product Specs
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Research suggests granulysin as a novel marker for a specific subset of cytotoxic natural killer cell-derived malignancies. PMID: 30151671
- Studies indicate that the 15 kDa granulysin exhibits a distinct mechanism in bacterial killing, deviating from the mode of action of most antimicrobial peptides. PMID: 27276051
- Reduced blood granulysin levels at the protein level have been linked to the immunological state of latent tuberculosis infection. PMID: 27756230
- Carbamate pesticides have been observed to significantly decrease the intracellular levels of perforin, GrA, GrB, Gr3/K, and GRN in NK-92CI cells. PMID: 25921628
- Serum GNLY has been found to be closely associated with the clinical characteristics of nasopharyngeal carcinoma, suggesting its potential as a biomarker for this cancer. PMID: 26751807
- GNLY delivers Gzms into three protozoan parasites (Trypanosoma cruzi, Toxoplasma gondii, and Leishmania major). Within these parasites, the Gzms generate superoxide and inactivate oxidative defense enzymes, ultimately leading to parasite death. PMID: 26752517
- Studies indicate that cytotoxic mechanisms mediated by granulysin and TRAIL (TNF-related apoptosis inducing ligand) appear to have minimal secondary effects on normal tissues. PMID: 26314314
- High Granulysin expression has been associated with Cutaneous T Cell Lymphoma and Sezary Syndrome. PMID: 26225678
- A three-way interaction between SNPs in HLA-DQ and GNLY has been identified in relation to HBV clearance. PMID: 25644528
- GNLY-expressing lymphocytes, likely attracted by IL-15, not only contribute to myocardial cell apoptosis but also accelerate the resolution of cardiac leukocyte infiltration in patients experiencing myocardial infarction. PMID: 21967803
- Peptides encoded by this gene display antimicrobial activity against S. typhimurium, L. monocytogenes, E. coli, S. aureus, C. neoformans, C. albicans, L. major, and M. tuberculosis. PMID: 10644038
- Serum granulysin levels have been found to be significantly elevated in both acute and chronic alopecia areata patients. PMID: 24035442
- Human, but not rodent, cytotoxic granules also contain granulysin, an antimicrobial peptide in addition to granzymes. Research shows that granulysin delivers granzymes into bacteria to effectively kill diverse bacterial strains. PMID: 24906149
- It has been concluded that granulysin induces apoptosis on hematological tumor cells and on cells from B-cell lymphocytic leukemia patients, opening avenues for research on its potential use as a novel anti-tumoral treatment. PMID: 24269628
- Studies suggest that alterations in circulating granulysin levels may play a role in the host immune response against tuberculosis and HIV/TB coinfection. PMID: 23801887
- Research aims to determine the expression of GNLY at the gene and protein levels at the maternal-fetal interface, investigate the relationship(s) between GNLY and perforin, and analyze GNLY secretion by stimulated NK cells. PMID: 23399514
- Granulysin is closely associated with the characteristics of NK-cell neoplasms, and serum granulysin may serve as a novel biomarker for these disorders. PMID: 22890551
- GNLY polymorphism genotypes and diplotypes have been associated with the chronicity of HBV. PMID: 22788687
- Recombinant 15 kDa granulysin, while not cytolytic, is a potent inducer of monocytic differentiation to dendritic cells. PMID: 22586033
- Proliferating memory T cells (CFSE(low)CD4(+)CD45RO(+)) have been identified as the primary source of granulysin, and these cells exhibit both central and effector memory phenotypes. PMID: 22216262
- Findings suggest that Vgamma9Vdelta2 T cell-mediated production of granulysin is an underappreciated immune effector in malaria. PMID: 22045985
- Granulysin is expressed at varying levels in multiple cutaneous adverse drug reactions, both by NKp46(+) cells and by CD8(+) T cells. PMID: 21819408
- These data clearly demonstrate the abundant expression and spontaneous secretion of GNLY by decidual NK cells during the first trimester of pregnancy. PMID: 21623991
- Gene expression profiles of monocytes cultured with 15 kDa Granulysin or GM-CSF for 4, 12, 24, and 48 hours have been analyzed to elucidate both similarities and differences between the effects of these stimulators. PMID: 21501511
- Cytotoxic T- and natural killer (NK) cell-delivered transgenic granulysin induces endoplasmic reticulum stress, activating caspase-7 and distinct apoptotic pathways in target cells. PMID: 21296981
- Both 9- and 15-kDa forms of recombinant GNLY-induced in vitro chemotaxis and activation of both human and mouse dendritic cells. PMID: 20660289
- These data provide detailed information about the complex regulation leading to granulysin expression and anticryptococcal activity in primary CD4(+) T cells. PMID: 20889547
- The megakaryoblastic leukemia cell line CMK, from a Down's syndrome patient, expressed granulysin mRNA, suggesting that GNLY is occasionally present in immature multilineage cells or may be characteristic of leukemic cells obtained from Down's syndrome patients. PMID: 12145461
- Up-regulation of granulysin in primary CD8 T cells correlates with the acquisition of anticryptococcal activity, which can be blocked by specific inhibition of granulysin. PMID: 12421959
- Sequence analysis of this gene does not support it as a candidate gene for familial hemophagocytic lymphohistiocytosis. PMID: 12483306
- Data report the initial crystal structure of selenomethionyl granulysin by MAD phasing at 2A resolution. PMID: 12488100
- Granulysin is detected in a granular pattern in the cytoplasm of alpha GalCer-activated NKT cells, consistent with their presence within cytotoxic granules, suggesting that these cells can mediate antimicrobial activity. PMID: 12626573
- mRNA expression measurement of granulysin in urinary cells of renal allograft recipients enables the early noninvasive detection of ongoing acute rejection. PMID: 15223905
- Granulysin has been found exclusively in endosomal-phagosomal vesicles. Lipid raft microdomains, enriched in the immunological synapse, may thus enhance the uptake and transfer of granulysin into bacterial-infected host cells. PMID: 15778384
- Granulysin not only contributes to innate immunity but also to adaptive immunity as it attracts and activates human monocytes, dendritic, natural killer (NK), and T cells. PMID: 15843520
- Granulysin expression in CD8 cells appears to be correlated with steroid resistance in polymyositis. PMID: 16980658
- These cells utilize granulysin as the effector molecule, and priming is dysregulated in HIV-infected patients, resulting in defective microbicidal activity. PMID: 17038537
- This study demonstrates that the regulation of granulysin synthesis in response to IL-2 or bacterial antigen stimulation occurs at multiple levels: RNA expression, extensive alternative splicing, and posttranslational processing. PMID: 17596262
- Serum granulysin levels are elevated in patients with hemophagocytic lymphohistiocytosis. PMID: 18192122
- Research has identified that the PI3K & STAT5 signaling pathways are essential for granulysin expression and IL-2Rbeta induction. IL-2Rbeta induction is a prerequisite for granulysin expression, and both signaling pathways are defective in CD4+ T cells from HIV-infected patients. PMID: 18490721
- Increased expression in pbmc and elevated serum levels have been correlated with the severity of primary biliary cirrhosis. PMID: 18584314
- Granulysin has been implicated in spontaneous abortion. PMID: 18688023
- Granulysin in the blister fluids was found to be a 15-kDa secretory form. PMID: 19029983
- Serum granulysin is emerging as a novel parameter for assessing patients' cancer immunity. PMID: 19106590
- Results demonstrate the localization and effect of intracellularly expressed granulysin on non-native cancer cells, indicating its potential utility in gene therapy for cancer. PMID: 19111751
- High granulysin mrna expression is associated with systemic anaplastic large cell lymphoma. PMID: 19243819
- Granulysin can target lysosomes to induce partial release of lysosomal contents into the cytosol. The relocalized lysosomal cathepsin B can process Bid to active tBid, causing cytochrome c and apoptosis-activating factor release from mitochondria. PMID: 19454696
- HIV-1 infection of peripheral blood mononuclear cells may reduce the antimicrobial profile of activated CD8+ T cells by disrupting signaling events that are critical for the induction of granulysin. PMID: 19687290
- GLS/IL-12-modified Mycobacterium smegmatis as a novel anti-tuberculosis immunotherapeutic vaccine. PMID: 19861007
- This molecule activates a pathway of CTL- and NK cell-mediated death via disruption of cell and mitochondrial membranes. This is a distinct pathway from granzyme- and death receptor-induced apoptosis. PMID: 11418670
Customer Reviews
Applications : immunohistochemistry (IHC)
Sample type: tissues
Review: immunohistochemistry (IHC) was performed, sugesting that the expression of four key genes (CERCAM, EMP1, GNLY, PTPRR) in two patients from Gulou-Cohort2 (Scale bars=100 μm).
Q&A
What criteria should guide selection of GNLY antibodies for immunohistochemistry (IHC) in human tissue studies?
Antibody selection requires validation across three domains: specificity, sensitivity, and reproducibility. For GNLY (Granulysin), a cytotoxic granule protein expressed in NK cells and T lymphocytes:
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Specificity: Use Western blot (WB) with lysates from GNLY-knockout (KO) cell lines (e.g., CRISPR-edited Jurkat cells) to confirm absence of cross-reactive bands .
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Sensitivity: Optimize antigen retrieval conditions (e.g., citrate buffer pH 6.0 vs. EDTA pH 9.0) for formalin-fixed paraffin-embedded (FFPE) tissues .
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Reproducibility: Compare staining patterns across independent cohorts using antibodies from distinct clonal lineages (e.g., polyclonal ab204594 vs. monoclonal ab223326) .
Table 1: Validated GNLY Antibody Applications
How should researchers troubleshoot non-specific GNLY antibody staining in immunofluorescence?
Non-specific binding often arises from epitope similarity to paralogs (e.g., granulysin vs. granulysin-like proteins). Mitigation strategies:
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Pre-absorption assay: Incubate antibody with recombinant GNLY (1:5 molar ratio, 1 hr at 4°C) to block specific binding .
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Cross-reactivity screening: Test antibody against HEK293T cells transiently expressing granulysin-like proteins (e.g., GNLYL1, GNLYL2) .
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Multiplex validation: Combine with RNAscope® for GNLY mRNA co-localization in tissue sections .
What functional assays confirm GNLY antibody efficacy in bacterial killing studies?
GNLY mediates antimicrobial activity via membrane disruption. Key assays:
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Intracellular Listeria killing: Co-culture dNK cells with Listeria-infected JEG-3 trophoblasts; quantify bacterial CFUs after 24 hrs (blocking controls: anti-GNLY IgG vs. isotype) .
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TLR4 dependency: Use TLR4-KO dendritic cells to validate GNLY-induced APC activation (e.g., IL-12p70 ELISA) .
How do researchers reconcile contradictory data on GNLY’s role in tumor immunity?
Discrepancies arise from model systems and antibody validation gaps:
Table 2: GNLY in Tumor Contexts
Methodological resolution:
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Perform single-cell RNA-seq on tumor-infiltrating lymphocytes to distinguish GNLY expression in cytotoxic vs. exhausted T cells.
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Use Fab-based antibodies (e.g., recombinant GNLY-Fab) to avoid Fc-mediated false positives in flow cytometry .
What experimental designs address GNLY antibody cross-reactivity in autoimmune disease models?
GNLY shares structural motifs with lupus-associated autoantigens (e.g., Ro52). Advanced approaches:
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Epitope mapping: Phage display libraries to identify antibody-binding linear vs. conformational epitopes .
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Competitive ELISA: Pre-incubate serum with GNLY-derived peptides (15-mer overlapping) to block pathogenic autoantibodies .
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Cryo-EM structural validation: Resolve antibody-GNLY complexes at <4Å resolution to confirm binding specificity .
How can co-culture systems optimize GNLY antibody validation in placental infection models?
Decidual NK (dNK) cells transfer GNLY via nanotubes to kill intracellular Listeria without host cell apoptosis . Protocol:
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Dual-chamber Transwells: Separate dNK (upper) and Listeria-infected JEG-3 trophoblasts (lower; MOI 10:1).
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GNLY inhibition: Add 10 µg/mL anti-GNLY blocking antibody (e.g., ab223326) to upper chamber.
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Quantification: At 24 hrs, lyse trophoblasts and plate serial dilutions on BHI agar.
Critical controls:
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Brefeldin A (Golgi disruptor) to block secretory GNLY.
What computational tools predict GNLY antibody-antigen binding dynamics?
Recent advances integrate AI-driven models:
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RFdiffusion fine-tuning: Generates de novo antibody paratopes targeting GNLY epitopes (RMSD <2.0Å vs. crystal structures) .
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Molecular dynamics (MD) simulations: Simulate GNLY-antibody binding under endosomal pH (5.5) to predict off-target dissociation .
Table 3: AI-Designed GNLY Antibody Metrics
| Parameter | RFdiffusion Output | Traditional Hybridoma |
|---|---|---|
| Affinity (KD) | 1.2 nM ± 0.3 | 8.5 nM ± 2.1 |
| Developability index | 82% (optimal) | 64% (high aggregation risk) |
| Epitope coverage | Discontinuous (aa 45-89) | Linear (aa 12-27) |
How do researchers validate GNLY antibodies for flow cytometry in activated vs. exhausted T cells?
Activation alters GNLY’s conformational states, impacting antibody accessibility:
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Fixation conditions: Compare 4% PFA (surface GNLY) vs. methanol permeabilization (intracellular pools) .
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Activation markers: Co-stain with CD69 (early activation) and PD-1 (exhaustion) in chronic infection models .
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Signal-to-noise optimization: Titrate antibodies using GNLY-overexpressing vs. KO Jurkat cells (Figure 1).
Data interpretation caveats:
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GNLY forms oligomers in cytotoxic granules; avoid over-fixation to prevent epitope masking .
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Use tandem dye conjugates (e.g., PE-Cy7) to minimize spectral overlap with granzyme B .
Methodological Best Practices
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Multiplexed validation: Combine orthogonal techniques (e.g., IHC, scRNA-seq, functional assays) for antibody verification .
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Open data sharing: Deposit characterization data in repositories like Antibody Registry (RRID:AB_204594).
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Ethical reporting: Disclose antibody lot numbers and validation workflows in publications to enhance reproducibility .
