IL12B Antibody
Description
Structure and Function of IL12B
IL12B encodes the p40 subunit of IL-12, a cytokine produced by antigen-presenting cells (APCs) like dendritic cells and macrophages . The active IL-12 heterodimer comprises IL12A (p35) and IL12B (p40), which binds to the IL-12 receptor (IL-12Rβ1 and IL-12Rβ2) on T cells and natural killer (NK) cells to induce Th1 differentiation and IFN-γ production . IL12B also forms IL-23 (with IL23A) and IL-35 (with EBI3), mediating diverse immune responses .
Applications of IL12B Antibodies
IL12B antibodies are widely used in research and diagnostics to detect and quantify IL12B in tissues or biological fluids. Key applications include:
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Western Blotting (WB): Detecting IL12B in denatured protein samples .
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Immunohistochemistry (IHC): Identifying IL12B in paraffin-embedded tissue sections .
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Flow Cytometry (FC): Analyzing IL12B expression in immune cells .
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Enzyme-Linked Immunosorbent Assay (ELISA): Quantifying IL12B levels in serum or supernatants .
Clinical and Therapeutic Relevance
IL12B antibodies are primarily research tools but hold potential in diagnostics and therapy:
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Therapeutic Targeting: IL12B is part of IL-23 and IL-35, which are implicated in autoimmune diseases (e.g., psoriasis, multiple sclerosis) . Monoclonal antibodies targeting IL12B/p40 (e.g., ustekinumab) inhibit IL-12/IL-23 signaling, reducing inflammation .
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Cancer Immunotherapy: Tumor-targeted IL-12 (e.g., NHS-IL12) enhances antitumor immunity while minimizing systemic toxicity . Antibodies like ebdarokimab (APR10252) are under development for IL12B modulation .
Research Insights
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Immune Regulation: IL12B antibodies enable studies of Th1/Th17 responses and chronic infections (e.g., mycobacteria, Salmonella) .
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Inflammatory Bowel Disease (IBD): Genetic variants in IL12B (e.g., rs6887695) correlate with IBD susceptibility . Antibodies aid in validating these associations in patient cohorts.
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Multiple Sclerosis: Overexpression of IL12B in the CNS suggests its role in MS pathogenesis, with antibodies used to map cytokine distribution .
Product Specs
Target Background
- Effect of polymorphisms in IL-12B p40, IL-17A and IL-23 A/G genes on the response of psoriatic patients to narrowband UVB. PMID: 29763989
- Research indicates that IL-12B A1188C (rs3212227) may contribute to the progression of the disease in the Czech population. PMID: 30069682
- Studies have confirmed an association between IL12B and IL23R genetic polymorphism and psoriasis vulgaris, suggesting a protective effect of minor alleles. PMID: 29454820
- The IL-12 B gene AA genotype has been observed more frequently in late stages of Hepatitis C virus chronically ill patients, while the CC genotype had no significant association with staging of liver disease and was less prevalent in late stages of liver disease. PMID: 28595541
- The frequency of the IL-12B AA (rs6871626) genotype was increased in Ankylosing Spondylitis patients. This suggests that IL-12B AA (rs6871626) gene polymorphisms could serve as potential biomarkers for diagnosis and prognosis in Ankylosing Spondylitis patients. PMID: 29200018
- The demonstrated allelic expression imbalance suggests that the IL12B risk haplotype confers susceptibility not only to Crohn's disease onset but also to relapse through increased IL12B mRNA expression. PMID: 28229296
- This study investigated the IL-12B gene status in 50 new sputum smear-positive pulmonary tuberculosis patients. PMID: 28697396
- The rs17860508 polymorphism in the IL12B promoter region may influence the risk of developing ovarian endometriosis by altering the endometrial expression of IL12B in Northern Chinese women. PMID: 29738836
- The rs6871626 single nucleotide polymorphism in IL-12B is associated with the pathophysiology of Takayasu arteritis. PMID: 28874185
- Human neutrophil elastase is involved in transactivation of TLR4 through activation of DUOX-2/EGFR and synergistically enhances IL-12p40 production by macrophages stimulated with LPS. PMID: 27282560
- Research suggests a significant association between IL-12B 3'-UTR and rs6887695 SNPs and Alzheimer's disease (ADs). PMID: 27068848
- Single-nucleotide-polymorphisms of IL12B may be considered a high-risk factor for Takayasu arteritis in the Chinese Han population. PMID: 28160070
- The IL-12B -1188 C allele may be a protective factor against asthma in East Asians. PMID: 28287286
- Low expression of HDAC3 and overexpression of inflammatory cytokines (IL-18, IL-12 and TNF-alpha) in intrahepatic cholestasis of pregnancy may be involved in liver cell apoptosis and the pathophysiology of the disease. PMID: 28697498
- PARP-1 has been identified as a unique regulator of Il12b transcription in response to inflammatory insults in an allele-differentiating manner. PMID: 28219892
- Pre-incubation with IL-12 was able to restore IFN-gamma production and the cytotoxic capacities of NK cells. This cytokine may therefore be considered as a potential treatment candidate in traumatic brain injury patients with immune suppression. PMID: 26387630
- Visceral leishmaniasis has been reported in two patients with IL-12p40 and IL-12Rbeta1 deficiencies. PMID: 27873456
- NOD2 rs3135500 and IL12B rs1368439 SNPs were not identified as genetic risk factors for colorectal cancer in the studied Iranian population. PMID: 27426943
- While IL-12B gene polymorphisms were not associated with rheumatoid arthritis (RA), rs6887695 was associated with RA in Asian patients. PMID: 27155343
- Research suggests that Il12B SNPs could be a genetic marker of disease severity in Takayasu arteritis. PMID: 25783557
- The results of this study suggested that rs6887695 SNP in the IL12B gene may increase susceptibility to relapsing-remitting multiple sclerosis. PMID: 28276258
- Meta-analyses have demonstrated that IL-12B rs3212227 and rs6887695 polymorphisms do not confer susceptibility to rheumatoid arthritis. PMID: 27312970
- Findings indicate that IL-12p40 + 1188A/C polymorphism as well as IL-12p70 protein levels may be associated with rheumatoid arthritis (RA) in the Polish population. PMID: 27896842
- The present meta-analysis suggests that the IL12B +1188A/C (rs3212227) polymorphism might be associated with genetic susceptibility to autoimmune diseases, such as type 1 diabetes, rheumatoid arthritis, and Behcet disease, but not Graves disease and ankylosing spondylitis. PMID: 26915668
- This study shows that IL-12B gene polymorphism does not contribute to the risk of hepatocellular carcinoma on top of chronic hepatitis C virus infection in Egyptian patients. PMID: 27819525
- This study has demonstrated (i) a significant association of -1082GG IL-10 genotype with susceptibility to autoimmune thyroid disease and (ii) the concomitant presence of IL-12B +1188CC and IL-10-1082 GG genotype contributes to the development and progression of Hashimoto's thyroiditis. PMID: 27774749
- Il-12B SNPs have an association with anti-HBs Ab production after vaccination in healthy Korean infants. PMID: 28051794
- Our results reveal that the IL12A rs568408 variant may be a marker SNP for the risk of both HBV clearance and HBV-related hepatocellular carcinoma development. PMID: 26631030
- This study shows that the Il-12 genetic polymorphisms are associated with susceptibility to tuberculosis in patients and their household contacts in India. PMID: 27108964
- High IL12 levels can inhibit Candida albicans colonizing the gastrointestinal tract of children and adolescents with diabetes mellitus type 1. PMID: 28127111
- This study shows that IL-12B gene polymorphism is a genetic risk factor for systemic lupus erythematosus in the Polish population, and that it predicts disease phenotype. PMID: 27059274
- Analysis of cytokine gene polymorphisms suggests that the IL-12 gene may play a role in the specific pathogenesis of ophthalmopathy in Korean children with early onset autoimmune thyroid disease. PMID: 26850223
- IL-12 polymorphism rs3212227 might not be a critical risk factor for Preeclampsia in Chinese Han women. PMID: 27148908
- A tendency of a higher prevalence of the genotype IL-12p40 pro1.1 in systemic arthritis and rheumatoid factor negative polyarthritis was observed but was not statistically significant. PMID: 26667304
- Genetic polymorphism has been associated with psoriasis in South Indian Tamils. PMID: 26472011
- Genetic polymorphism has been associated with allergic rhinitis in Chinese patients. PMID: 26663019
- The IL23R polymorphisms rs10889677, rs7517847, and the IL12B polymorphism rs3212227 are not associated with multiple sclerosis risk. PMID: 26000455
- IL-12 immunomodulation delays the onset of lethal peritoneal disease of ovarian cancer. PMID: 26430781
- Variant IL-12p40 1188C/A genotype was observed as AA (72.92%), AC (23.96%), and CC (3.13%) in patients compared to 65%, 30%, and 5%, respectively, in controls. PMID: 26516307
- The IL-12B rs3212227 AC and AC+CC genotypes are associated with rheumatoid arthritis risk in older patients, rheumatoid factor-positive patients, and anti-cyclic peptide antibodies-negative patients. PMID: 26375522
- High serum IL-12 levels associated with increased densities of peritumoral CD8(+) T cells, intraepithelial CD3(+) T cells, and intratumoral neutrophils, while high serum CCL4 levels associated with increased densities of peritumoral CD68(+) cells. PMID: 26874795
- These observations indicate a HDAC1-mediated IL-12B gene expression suppression by live, virulent Mycobacterium tuberculosis to subvert the immune system to survive and replicate in the host. PMID: 26697414
- IL-12p40 response to in vitro stimulation in patients with Mendelian susceptibility to mycobacterial disease was below normal levels. PMID: 25201764
- Eight SNPs (rs10045431, rs11167764, rs3212227, rs6556412, rs6556416, rs6871626, rs6887695 and rs7709212) were genotyped in Han ankylosing spondylitis patients and controls. rs6871626 may be associated with AS susceptibility and activity. PMID: 26103568
- Replicating the association of single-nucleotide polymorphisms in the TNFAIP3, IL12B, and IL23R genes with psoriasis vulgaris in subjects from different ethnic backgrounds underlines their importance in the pathogenesis of the disease. PMID: 25406098
- The balance of IL-23 versus IL-12/IL-27 signals into CD4(+) effector T cells determines whether tissue inflammation is perpetuated or resolves. PMID: 24796719
- Compared with the IL-12B-AA genotype, CC and combined CC/AC genotypes were associated with a significantly decreased risk of Hepatitis C virus infection in Chinese hemodialysis patients. PMID: 25613737
- These results show that the IL-15 and IL-12 combination has the ability to expand the selective depletion of invariant natural killer T cells in vitro in HIV-infected individuals. PMID: 24748538
- The risk of gastric cardiac adenocarcinoma associated with the IL12B rs3212227 T > G polymorphism was evident among Chinese female patients and Chinese patients who never smoked or consumed alcoholic drinks. PMID: 24529168
- Based on these results, we believe that increased levels of IL-12p40 and IL-16 are associated with an ongoing inflammatory response in obese individuals and could lead to the development of disease conditions related to obesity. PMID: 25710036
Q&A
What structural features of IL12B are critical for antibody binding and functional assays?
IL12B (p40) forms a disulfide-linked heterodimer with IL12A (p35) to create bioactive IL12 (p70). Antibodies targeting IL12B must recognize epitopes that do not interfere with IL12 receptor (IL12Rβ1/β2) binding or heterodimer stability. Key structural considerations include:
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Glycosylation sites: IL12B contains N-linked glycans at Asn222 and Asn395, which influence antibody affinity. Deglycosylation assays (e.g., PNGase F treatment) can confirm epitope dependence on glycosylation .
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Domain specificity: The p40 subunit includes a conserved IL12B_N domain (residues 23-200) and a cytokine-binding domain (residues 201-328). Antibodies targeting the cytokine-binding domain may block IL12-IL12R interactions, while those binding IL12B_N are useful for Western blotting .
Table 1: Common IL12B Antibody Validation Assays
How do researchers validate IL12B antibody specificity across experimental models?
Cross-reactivity with IL23 (which shares the p40 subunit) is a major challenge. A three-step validation framework is recommended:
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Epitope mapping: Use truncated IL12B variants or alanine-scanning mutagenesis to identify critical binding residues. For example, antibodies targeting residues 180-200 of IL12B show no cross-reactivity with IL23 .
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Functional blocking: Assess inhibition of IL12-dependent IFNγ production in phytohemagglutinin-activated PBMCs. Effective blocking antibodies reduce IFNγ by ≥80% at 1 µg/mL .
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Multi-platform consistency: Compare results across ELISA, Western blot, and immunohistochemistry. Discordant signals often indicate post-translational modification effects .
What controls are essential for quantifying IL12B in complex biological matrices (e.g., serum)?
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Pre-analytical controls: EDTA plasma is preferred over serum to prevent platelet-derived IL12B release during clotting .
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Matrix interference controls: Spike-and-recovery experiments using recombinant IL12B (70–95 kDa) at 0.1–10 ng/mL in diluted plasma (1:4 in PBS) account for nonspecific binding .
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Reference standards: Calibrate assays against WHO International Standard for IL12 (NIBSC code 12/170) to ensure inter-study reproducibility .
How should researchers design experiments to resolve contradictions in IL12B expression data across studies?
Conflicting reports on IL12B levels often stem from differences in:
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Sample timing: IL12B peaks 36–48 hours post-immune stimulation (e.g., BCG infection) and declines by day 7 .
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Disease heterogeneity: In cancer trials, responders show transient IL12B induction (peak at 24 h), while non-responders exhibit sustained elevation (>72 h) .
Table 2: IL12B Expression Variability in Clinical Cohorts
Methodological recommendations:
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Stratify patients by IL12B genotype (e.g., g.315_316insA frameshift vs. wild-type) .
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Use longitudinal sampling (0, 24, 48, 72 h) to capture kinetic profiles .
What statistical approaches are optimal for analyzing IL12B-mediated cytokine networks?
In a study of 21 cancer patients, IL12B administration induced divergent cytokine patterns:
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Responders: GM-CSF↑ (2.1-fold, p<0.01), VEGF-C↓ (1.8-fold, p=0.03) .
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Non-responders: CCL3↑ (4.3-fold), IL-6↑ (3.7-fold), IFNγ↑ (2.9-fold) (p<0.001) .
Analysis pipeline:
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Dimensionality reduction: Principal component analysis (PCA) to cluster patients by cytokine trajectories.
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Causal inference: Structural equation modeling (SEM) to identify IL12B-driven vs. bystander cytokine effects.
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Time-series modeling: Generalized additive models (GAMs) to account for non-linear pharmacokinetic/pharmacodynamic relationships .
How can IL12B antibody studies inform dose escalation in early-phase clinical trials?
Data from the NHS-IL12 trial (N=59) revealed:
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MTD: 16.8 µg/kg every 4 weeks, with dose-limiting toxicities (DLTs) including grade 3 transaminitis .
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Pharmacokinetics: Terminal half-life = 48–72 h, supporting biweekly dosing .
Table 3: Clinical Trial Design Considerations for IL12B-Targeted Agents
| Parameter | Recommendation | Rationale |
|---|---|---|
| Starting dose | 2 µg/kg | 1/10 of NOAEL in primates |
| Biomarker sampling | Days 1, 2, 3, 7, 14 | Capture IFNγ/IL10 surge and rebound |
| Response assessment | Modified irRC at 8-week intervals | Account for pseudoprogression |
Troubleshooting tip: Pre-dose corticosteroids (e.g., dexamethasone 4 mg) attenuate IL12B-induced cytokine release syndrome without compromising antitumor efficacy .
