CCL22/MDC Antibody (57226) [Unconjugated]

Recombinant Human CCL22/MDC (Catalog # 336-MD) chemoattracts the BaF3 mouse pro‑B cell line transfected with human CCR4 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Human CCL22/MDC (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human CCL22/MDC Monoclonal Antibody (Catalog # MAB336). The ND50 is typically 0.6-3.0 µg/mL.

MiR‐23a inhibitor reversed p65 inhibition effects on HBV‐positive xenograft tumor growth. A, Xenografted tumors were harvested after different treatments. B and C, p65 inhibitor treatment significantly retarded tumor growth and miR‐23a inhibitor promoted tumor growth as indicated by the tumor weight and size. The therapeutic effects of p65 inhibitor were abolished by cotreatment with miR‐23a inhibitor. D, CCL22 protein levels from each group were detected by western blotting. E, Statistical results in D. p65 inhibitor led to reduction of CCL22 protein level but miR‐23a inhibitor increased CCL22 level in tumor tissues. F, p65 inhibitor led to reduction of CCL22 mRNA level but miR‐23a inhibitor increased CCL22 mRNA level in tumor tissues. MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 level in tumor tissues. Error bars represented mean ± SD. *P < .05 and **P < .01 Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.

MiR‐23a, p‐p65, p65, and CCL22 levels were dysregulated in HCC cell lines. A, RT‐qPCR revealed that miR‐23a expression was lowest in the HBV‐positive HepG2.2.15 cell line. B, Highest mRNA level of CCL22 was observed in HepG2.2.15. C, Protein levels of p‐p65, p65, and CCL22 in three cell lines were determined by Western blotting. D, The gray scale analysis of p‐p65, p65, CCL22 and ratio of p‐p65/p65 in three cell lines. Expression of CCL22, p‐p65, and p65 were higher in HBV+ HepG2.2.15 cells than their parental HBV‐ HepG2 cells and normal WRL68 cells. Error bars represented mean ± SD. **P < .01 and *P < .05 Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.

MiR‐23a inhibited Tregs migration by directly targeting CCL22. A, CCL22 3′UTR contains a putative miR‐23a‐binding site. B, MiR‐23a mimics significantly reduced the luciferase activity tagged with wide type of CCL22 3′UTR. Mutagenesis in the miR‐23a‐binding site abolished the inhibitory actions. C, Transfection of miR‐23a mimics efficiently increased miR‐23a level in both cell lines. D, The effects of miR‐23a overexpression on p‐p65, p65, and CCL22 protein levels in HepG2 and HepG2.2.15 cells were assessed by western blotting. E, The gray scale analysis of p‐p65, p65, CCL22 after miR‐23a overexpression. MiR‐23a mimics transfection did not alter the expression of p65 and its phosphorylation but CCL22 was significantly reduced by miR‐23a mimics. F, MiR‐23a mimics significantly reduced CCL22 mRNA level. G, Overexpression of miR‐23a was sufficient to attenuate Tregs transmigration in both cell lines. Error bars represented mean ± SD. *P < .05, **P < .01, and ***P < .001 Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.

Lower miR‐23a expression was correlated with higher level of CCL22 expression and intratumoral Treg recruitment in HBV‐positive HCC. A, HBV infection was associated with patient survival rate of HCC. Patients carrying HBV (n = 30) had significantly poorer prognosis than HBV‐ patients (n = 30). B, HBV‐ tissues (n = 30) and HBV+ tissues (n = 30) expressed significantly lower level of miR‐23a. C, HBV− tissues and HBV+ tissues expressed significantly higher mRNA level of CCL22. D, HBV‐ tissues and HBV+ tissues expressed significantly higher mRNA level of Foxp3. E, The protein levels of CCL22, Foxp3, p‐p65, and p65 in normal control, HBV‐ and HBV+ tumor tissues were evaluated by western blotting. F, The gray scale analysis of CCL22, Foxp3, p‐p65, and p65 in normal control, HBV− and HBV+ tumor tissues. In ascending order: normal < HBV− < HBV+. G, Foxp3 signals (red) were colocalized with CD4 (green) signals in tissues. The ratios of Foxp3+CD4+ cells were gradually increased from normal, HBV− HCC to HBV+ HCC tissues. H. MiR‐23a level was inversely correlated with CCL22 expression in HCC tissues, but not in normal control. I. MiR‐23a level was inversely correlated with Foxp3 expression in HCC tissues, but not in normal control. Error bars represented mean ± SD. **P < .01 and *P < .05. HBV+, HCC tissues with HBV infection. HBV−, HCC tissues without HBV infection. Normal, normal liver samples Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.

MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 and Tregs recruitment. A, MiR‐23a inhibitor reversed p65 inhibition effects on miR‐23a level. B, The effects of miR‐23a inhibition and p65 inhibitor treatment on p‐p65, p65, and CCL22 protein levels in HepG2 and HepG2.2.15 cells were assessed by western blotting. C, The gray scale analysis of p‐p65, p65, CCL22. MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 protein level but had no effects on p‐p65 and p65. D, MiR‐23a inhibitor reversed p65 inhibition effects on CCL22 mRNA level. E, MiR‐23a inhibitor reversed p65 inhibition effects on Tregs recruitment. F, CCL22 neutralizing antibody reversed miR‐23a inhibitor‐mediated Tregs migration. Error bars represented mean ± SD. *P < .05, **P < .01, and ***P < .001 Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.

p65 promoted Tregs recruitment via directly modulating miR‐23a. A, A p65‐binding site could be found in miR‐23a promoter region. B, Overexpression of p65 significantly reduced the luciferase activity driven by miR‐23a promoter. Mutagenesis in the p65‐binding site released such repression. C, ChIP assay confirmed that p65 could bind to the promoter region of miR‐23a. D, p65 inhibition upregulated miR‐23a level. E, p65 inhibitor significantly reduced the p‐p65, p65, and CCL22 protein levels in HepG2 and HepG2.2.15 cells. F, The gray scale analysis of p‐p65, p65, CCL22 after p65 inhibitor treatment. G, p65 inhibition dampened CCL22 mRNA level. H, CD4+CD25+ Foxp3+ human Tregs were identified. I, HBV+ HepG2.2.15 cells caused more Tregs to migrate through transwell membrane than HBV− HpeG2 cells. Pretreatment with p65 inhibitor attenuated Tregs transmigration in both cell lines. Error bars represented mean ± SD. *P < .05, **P < .01, and ***P < .001 Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31769216), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, B/N, ELISA(Cap)
• Clone: 57226
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

CCL22/MDC Antibody (57226) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human CCL22/MDC
  Gly25-Gln93
  Accession # O00626.1
• Specificity: Detects human CCL22/MDC in ELISAs and Western blots. In ELISAs, no cross-reactivity with recombinant mouse CCL17, recombinant human (rh) CCL17, or rhCCL21 is observed.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: CCL22
• Purity Statement: Protein A or G purified from ascites
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • ELISA Capture (Matched Antibody Pair) 2-8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.1-0.4 ug/mL
  • ELISA Standard (Matched Pair)
  • Neutralization 0.6-3.0 ug/mL
  • Western Blot 1 ug/mL
• Application Notes: ELISA Detection: Human CCL22/MDC Biotinylated Antibody (Catalog number BAF336)
  Standard: Recombinant Human CCL22/MDC (Catalog number 336-MD)

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CCL22/MDC Antibody (57226) [Unconjugated]

• A-152E5.1
• ABCD-1
• CC chemokine STCP-1
• C-C motif chemokine 22
• CCL22
• chemokine (C-C motif) ligand 22
• DC/B-CK
• Macrophage-derived chemokine
• MDC
• MDCStimulated T-cell chemotactic protein 1
• MGC34554
• SCYA22MDC(1-69)
• small inducible cytokine A22
• small inducible cytokine subfamily A (Cys-Cys), member 22
• Small-inducible cytokine A22
• STCP-1
• stimulated T cell chemotactic protein 1

Background

CCL22, also named stimulated T cell chemotactic protein (STCP-1) and MDC, is a CC chemokine initially isolated from clones of monocyte-derived macrophages. Human CCL22 cDNA encodes a precursor protein of 93 amino acid residues with a 24 amino acid residue predicted signal peptide that is cleaved to yield a 69 amino acid residue mature 8 kDa protein. At the amino acid sequence level, CCL22 shows less than 35% identity to other CC chemokine family members. Human CCL22 is expressed in dendritic cells, macrophages and activated monocytes. In addition, CCL22 expression is also detected in the tissues of thymus, lymph node and appendix. The gene for human CCL22 has been mapped to chromosome 16 rather than chromosome 17 where the genes for many human CC chemokines are clustered. Recombinant or chemically synthesized mature CCL22 has been shown to induce chemotaxis or Ca²⁺ mobilization in dendritic cells, IL-2 activated NK cells, and activated T lymphocytes. A CD8⁺ T lymphocyte-derived secreted soluble activity that suppresses infection by primary non-syncytium-inducing and syncytium-inducing HIV-1 isolates and the T cell line-adapted isolate HIV-1_IIIB, has been identified as CCL22. Based on amino-terminal sequence analysis, the major CD8⁺ T lymphocyte-derived CCL22 protein yielded an amino-terminal sequence of YGANM, which is two amino acid residues shorter than the predicted mature CCL22. The difference in potency between the two mature CCL22 isoforms has not been determined.

1. Godiska, R. et al. (1997) J. Exp. Med. 185:1595.
2. Chang, M-S. et al. (1997) J. Biol. Chem. 272:25229.
3. Pal, R. et al. (1997) Science 278:5338.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CCL22/MDC Antibody (MAB336)(15)

Publications using MAB336

• Maroui, MA;Odongo, GA;Mundo, L;Manara, F;Mure, F;Fusil, F;Jay, A;Gheit, T;Michailidis, TM;Ferrara, D;Leoncini, L;Murray, P;Manet, E;Ohlmann, T;De Boevre, M;De Saeger, S;Cosset, FL;Lazzi, S;Accardi, R;Herceg, Z;Gruffat, H;Khoueiry, R; Aflatoxin B1 and Epstein-Barr virus-induced CCL22 expression stimulates B cell infection Proceedings of the National Academy of Sciences of the United States of America 2024-04-16 [PMID: 38574017] (Western Blot, Neutralization, Human, Transgenic Mouse)
• C Kötting, L Hofmann, R Lotfi, D Engelhardt, S Laban, PJ Schuler, TK Hoffmann, C Brunner, MN Theodoraki Immune-Stimulatory Effects of Curcumin on the Tumor Microenvironment in Head and Neck Squamous Cell Carcinoma Cancers, 2021-03-16;13(6):. 2021-03-16 [PMID: 33809574] (ELISA Capture, Human)
• Xuan Zhao, Shasha Liu, Xinfeng Chen, Jianyi Zhao, Feng Li, Qitai Zhao et al. L1CAM overexpression promotes tumor progression through recruitment of regulatory T cells in esophageal carcinoma Cancer Biology and Medicine 2021-05-15 [PMID: 33710805]
• ZQ Li, HY Wang, QL Zeng, JY Yan, YS Hu, H Li, ZJ Yu p65/miR-23a/CCL22 axis regulated regulatory T cells recruitment in hepatitis B virus positive hepatocellular carcinoma Cancer Med, 2019-11-25;0(0):. 2019-11-25 [PMID: 31769216] (Western Blot, Human)
• Alejandro L. Antonia, Kyle D. Gibbs, Esme D. Trahair, Kelly J. Pittman, Amelia T. Martin, Benjamin H. Schott et al. Pathogen Evasion of Chemokine Response Through Suppression of CXCL10 Frontiers in Cellular and Infection Microbiology 8/7/2019 [PMID: 31440475]
• G Ren, JL Whittaker, C Leonard, D De Rantere, DSJ Pang, P Salo, M Fritzler, M Kapoor, APJ de Koning, JL Jaremko, CA Emery, RJ Krawetz CCL22 is a biomarker of cartilage injury and plays a functional role in chondrocyte apoptosis Cytokine, 2019-01-07;115(0):32-44. 2019-01-07 [PMID: 30623804] (IHC-P, Human)
• XW Yang, HX Jiang, R Lei, WS Lu, SH Tan, SY Qin Recruitment and significance of Th22 cells and Th17 cells in malignant ascites Oncol Lett, 2018-08-16;16(4):5389-5397. 8/16/2018 [PMID: 30250609]
• SH Omland, EE Wettergren, S Mollerup, M Asplund, T Mourier, AJ Hansen, R Gniadecki Cancer associated fibroblasts (CAFs) are activated in cutaneous basal cell carcinoma and in the peritumoural skin BMC Cancer, 2017-10-07;17(1):675. 10/7/2017 [PMID: 28987144]
• Oo YH, Weston CJ, Lalor PF, Curbishley SM, Withers DR, Reynolds GM, Shetty S, Harki J, Shaw JC, Eksteen B, Hubscher SG, Walker LS, Adams DH Distinct roles for CCR4 and CXCR3 in the recruitment and positioning of regulatory T cells in the inflamed human liver. J. Immunol., 2010-02-17;184(6):2886-98. 2010-02-17 [PMID: 20164417] (IHC-P, Human)
• Mizukami Y, Kono K, Kawaguchi Y, Akaike H, Kamimura K, Sugai H, Fujii H CCL17 and CCL22 chemokines within tumor microenvironment are related to accumulation of Foxp3+ regulatory T cells in gastric cancer. Int. J. Cancer, 2008-05-15;122(10):2286-93. 2008-05-15 [PMID: 18224687] (Flow Cytometry, Human)
• Haas J, Schopp L, Storch-Hagenlocher B, Fritzsching B, Jacobi C, Milkova L, Fritz B, Schwarz A, Suri-Payer E, Hensel M, Wildemann B Specific recruitment of regulatory T cells into the CSF in lymphomatous and carcinomatous meningitis. Blood, 2007-10-29;111(2):761-6. 2007-10-29 [PMID: 17967942] (Neutralization, Human)
• Liu LY, Bates ME, Jarjour NN, Busse WW, Bertics PJ, Kelly EA Generation of Th1 and Th2 chemokines by human eosinophils: evidence for a critical role of TNF-alpha. J. Immunol., 2007-10-01;179(7):4840-8. 2007-10-01 [PMID: 17878383] (ELISA Development, Human)
• Curiel , Tyler J, Coukos , George, Zou , Linhua, Alvarez , Xavier, Cheng , Pui, Mottram , Peter, Evdemon-Hogan , Melina, Conejo-Garcia , Jose R, Zhang , Lin, Burow , Matthew, Zhu , Yun, Wei , Shuang, Kryczek , Ilona, Daniel , Ben, Gordon , Alan, Myers , Leann, Lackner , Andrew, Disis , Mary L, Knutson , Keith L, Chen , Lieping, Zou , Weiping Specific recruitment of regulatory T cells in ovarian carcinoma fosters immune privilege and predicts reduced survival. Nat Med, 2004-08-22;10(9):942-9. 2004-08-22 [PMID: 15322536] (Neutralization, Human, Xenograft)
• Liu L, Jarjour NN, Busse WW, Kelly EA Enhanced generation of helper T type 1 and 2 chemokines in allergen-induced asthma. Am. J. Respir. Crit. Care Med., 2004-03-04;169(10):1118-24. 2004-03-04 [PMID: 15001464] (ELISA Development, Human)
• Conroy DM, Jopling LA, Lloyd CM, Hodge MR, Andrew DP, Williams TJ, Pease JE, Sabroe I CCR4 blockade does not inhibit allergic airways inflammation. J. Leukoc. Biol., 2003-07-15;74(4):558-63. 2003-07-15 [PMID: 12960268] (ELISA Development, Guinea Pig)

Bioinformatics

• Gene Symbol: CCL22
• Uniprot:
  • O00626.1()