human ccl22  (Proteintech)

Journal: Journal for Immunotherapy of Cancer

Article Title: Anti-HER2×CCR4 bispecific antibody enhances antitumor immunity in advanced HER2-positive tumors by chemotaxis blockade and depletion of tumor-associated Tregs, without inducing systemic toxicity

doi: 10.1136/jitc-2025-012829

Figure Lengend Snippet: The structures and properties of four anti-HER2×CCR4 DVD-Igs. ( A ) Anti-HER2×CCR4 DVD-Igs consist of two VD1 and two VD2. The VD1 and VD2 are connected by linkers of different lengths. ( B ) SDS-PAGE gels of purified proteins under NR and R conditions determined the molecular weights of the anti-HER2×CCR4 DVD-Ig. ( C ) The affinity of anti-HER2×CCR4 DVD-Igs to HER2 and CCR4 protein was determined by SPR. ( D, E ) The affinity of anti-HER2×CCR4 DVD-Igs to HER2 protein ( D ) and CCR4 protein ( E ) was determined by ELISA. ( F ) Competitive ELISA was used to assess the competition of CCL22 protein and antibodies in binding to CCR4 protein. ( G ) Flow cytometry analysis of HER2 expression in MDA-MB-231 (left), SKBR (middle), and NCI-N87 (right) cells. ( H ) Flow cytometry analyzes the binding of anti-HER2×CCR4 DVD-Igs to MDA-MB-231 (left), SKBR (middle), and NCI-N87 (right) cells. Representative flow cytometry plots. ( I ) Representative flow cytometry plots. ( J ) CCR4 expression on Treg-CCR4 cells was assessed by flow cytometry. ( K ) Flow cytometry analyzed the HER2×CCR4 DVD-Ig mediated co-binding of SKBR3 cells and Treg-CCR4 cells by calculating CFSE + PKH26 + populations. ( L ) Confocal microscopy was used to observe the co-binding of SKBR3 cells and Treg-CCR4 cells mediated by anti-HER2×CCR4 DVD-Ig. Green: HER2 + cells, red: CCR4 + cells. Microscope magnification: ×200 times. All in vitro experiments were performed three times independently. HER2, human epidermal growth factor receptor 2; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; SPR, surface plasmon resonance; Treg, regulatory T cell; VD, variable domains.

Article Snippet: Human CCL22 (CSB-AP000951HU, CUSABIO, https://www.cusabio.cn ) was coated onto plates.

Techniques: SDS Page, Purification, Enzyme-linked Immunosorbent Assay, Competitive ELISA, Binding Assay, Flow Cytometry, Expressing, Confocal Microscopy, Microscopy, In Vitro, Polyacrylamide Gel Electrophoresis, SPR Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Anti-HER2×CCR4 bispecific antibody enhances antitumor immunity in advanced HER2-positive tumors by chemotaxis blockade and depletion of tumor-associated Tregs, without inducing systemic toxicity

doi: 10.1136/jitc-2025-012829

Figure Lengend Snippet: XL-11 eliminates tumor-associated Tregs in vitro and prevents the exhaustion of Tregs in the peripheral circulation. ( A–C ) The in vitro ADCC assays of SKBR3 ( A ), NCI-N87 ( B ), and Treg-CCR4 cells ( C ). ( D ) Schematic of chemotaxis assay. ( E ) Anti-HER2×CCR4 DVD-Ig inhibited CCL22 chemotaxis to Treg-CCR4 + cells. ( F–I ) Teff cells were labeled with CFSE. The proportion of Teff cells was assessed by flow cytometry ( F, G ), and IFN-γ ( H ) and IL-2 ( J ) in the culture medium supernatant of the co-culture system was assessed by ELISA. ( J ) CD3 + CD4 + T cells from PBMC were isolated and differentiated to Tregs in vitro, and the proportion of CD3 + CD4 + T cells (left) and Tregs (middle, right) was assessed by flow cytometry. ( K ) The binding of anti-HER2×CCR4 DVD-Ig to Tregs was assessed by flow cytometry. All in vitro experiments were performed three times independently. Values were expressed as mean±SEM. ADCC, antibody-dependent cellular; CFSE, carboxyfluorescein succinimidyl ester; HER2, human epidermal growth factor receptor 2; IFN, interferon; IL, interleukin; PBMC, peripheral blood mononuclear cell; Teff, effector T cells; Treg, regulatory T cell.

Article Snippet: Human CCL22 (CSB-AP000951HU, CUSABIO, https://www.cusabio.cn ) was coated onto plates.

Techniques: In Vitro, Chemotaxis Assay, Labeling, Flow Cytometry, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Isolation, Binding Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Anti-HER2×CCR4 bispecific antibody enhances antitumor immunity in advanced HER2-positive tumors by chemotaxis blockade and depletion of tumor-associated Tregs, without inducing systemic toxicity

doi: 10.1136/jitc-2025-012829

Figure Lengend Snippet: A schematic illustrating the mechanism of XL-11-mediated antitumor activity without systemic toxicity. ( a ) CCR4 ligands CCL22 and CCL17 establish a chemotactic microenvironment around the tumor, recruiting peripheral CCR4 + Tregs into the TME. XL-11 inhibits CCL22/CCL17–CCR4 binding, suppressing Treg chemotaxis and infiltration into the TME. ( b ) XL-11 targets tumor cells and TIL-Tregs to eliminate these cells by inducing ADCC. This action overcomes TIL-Treg–mediated suppression and restores the function of Teff. Additionally, XL-11 prevents TIL-Treg migration to metastatic sites via CCR4 blockade. ( c ) XL-11 depletes tumor-chemotactic Tregs with high expression of CCR4 while maintaining Tregs essential for immune homeostasis in peripheral circulation. ADCC, antibody-dependent cellular cytotoxicity; ECM, extracellular matrix; HER2, human epidermal growth factor receptor 2; Teff, effector T cells; TIL, tumor-infiltrating lymphocyte; TME, tumor microenvironment; Treg, regulatory T cell.

Article Snippet: Human CCL22 (CSB-AP000951HU, CUSABIO, https://www.cusabio.cn ) was coated onto plates.

Techniques: Activity Assay, Binding Assay, Chemotaxis Assay, Migration, Expressing

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: rhCCL22-loaded PLGA microparticles exhibit different size distribution based on the manufacturing method. ( a , b ) Volume-weighted size distributions and ( c , d ) representative scanning electron microscopy images of rhCCL22-loaded PLGA microparticles that were prepared by ( a , c ) microfluidics or ( b , d ) conventional methods.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques: Electron Microscopy

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: Representative scanning electron microscopy images demonstrate the surface morphology and inner porosity differences for rhCCL22-loaded PLGA microparticles. PLGA microparticles were prepared by ( a , c ) microfluidics and ( b , d ) conventional methods. Arrows in ( d ) highlight some examples of the inner pore connections.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques: Electron Microscopy

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: rhCCL22 encapsulation (%) in the PLGA microparticles shows a significant difference depending on the manufacturing process: microfluidics vs. conventional methods. ** p ≤ 0.01.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques: Encapsulation

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: rhCCL22 release kinetics from PLGA microparticles demonstrates different release kinetics according to the manufacturing method. ( a ) Microparticles prepared by the microfluidics method showed a zero-order delayed (~2 days) release profile. ( b ) Microparticles prepared by the conventional method presented a zero-order release profile.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: Batch-to-batch variations in the size distribution of rhCCL22-loaded PLGA microparticles when PLGA microparticles were prepared by ( a ) microfluidics and ( b ) conventional methods.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: Batch-to-batch variations in terms of rhCCL22 encapsulation (%) in PLGA microparticles regarding the manufacturing method. rhCCL22-loaded PLGA microparticles were prepared by ( a ) microfluidics and ( b ) conventional methods. * p ≤ 0.05, *** p ≤ 0.001, ns indicates non-significant difference.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques: Encapsulation

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: Batch-to-batch variations in terms of rhCCL22 release kinetics from PLGA microparticles. rhCCL22-loaded PLGA microparticles were prepared by ( a ) microfluidics and ( b ) conventional methods.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques:

Journal: Translational Oncology

Article Title: CCL22 as an independent prognostic factor in endometrial cancer patients

doi: 10.1016/j.tranon.2024.102116

Figure Lengend Snippet: Representative images of CCL22-expression in EC: (A) intermediate expression in glandular cells (IRS=2.67), S/M (IRS=0.67); (B) high expression in glandular cells (IRS=12.00), S/M (IRS=5.33); (C) strongly positive cells in tumor distant S/M; (D) significant correlation between CCL22 in tumor- and stroma-cells; (E) CCL22 levels in tumor cells were higher than in S/M. Objective 20x, Scale bar 100 µm.

Article Snippet: Paraffin-embedded TMA of EC-patients and tissue of the control group were incubated with the polyclonal rabbit anti-human MDC (CCL22) antibody (500-P107 1:300, Peprotech) using ZytoChem Plus HRP Polymer System mouse/rabbit following the manufacturer's instructions.

Techniques: Expressing

Journal: Translational Oncology

Article Title: CCL22 as an independent prognostic factor in endometrial cancer patients

doi: 10.1016/j.tranon.2024.102116

Figure Lengend Snippet: Representative images of CCL22-expression in endometrial control with negative (A) and strong staining in glandular epithelial cells (B) and strongly positive cells in myometrium (C). Elevated CCL22-expression in glandular epithelial cells compared to stroma cells (D). Significantly lower CCL22-IRS EC compared to benign endometrium (E). Concerning CCL22 in S/M a trend to higher levels was found in the specimens of EC patients (F). Objective 20x, Scale bar 100 µm.

Article Snippet: Paraffin-embedded TMA of EC-patients and tissue of the control group were incubated with the polyclonal rabbit anti-human MDC (CCL22) antibody (500-P107 1:300, Peprotech) using ZytoChem Plus HRP Polymer System mouse/rabbit following the manufacturer's instructions.

Techniques: Expressing, Control, Staining

Journal: Translational Oncology

Article Title: CCL22 as an independent prognostic factor in endometrial cancer patients

doi: 10.1016/j.tranon.2024.102116

Figure Lengend Snippet: CCL22-expression in S/M increases with higher grade in EC (A). High CCL22-expression in S/M in EC is associated to poorer OS (B), but not to PFS (C). (D) High CCL22-expression in tumor epithelium is associated to prolonged OS, but not to PFS (E).

Article Snippet: Paraffin-embedded TMA of EC-patients and tissue of the control group were incubated with the polyclonal rabbit anti-human MDC (CCL22) antibody (500-P107 1:300, Peprotech) using ZytoChem Plus HRP Polymer System mouse/rabbit following the manufacturer's instructions.

Techniques: Expressing

Journal: Translational Oncology

Article Title: CCL22 as an independent prognostic factor in endometrial cancer patients

doi: 10.1016/j.tranon.2024.102116

Figure Lengend Snippet: Isolated CCL22+ cells were identified as mainly M1-macrophages in distant myometrial tissue areas by double immunofluorescence: (A) Representative EC stained for CCL22 (red) and CD68, CD80, CD163, and DEC205 (green); (B) Proportion of CCL22+ cells, that expressed also one of those immune cells markers is presented; (C) Proportion of total expression of all markers: dominant occurrence of CD68+ and CD80+ cells; (D) Kaplan-Meier Curve for OS and PFS (E). Correlation analysis of FoxP3 and CCL22 in S/M (F), distant strongly positive cells (G), and in tumor cells (H).

Article Snippet: Paraffin-embedded TMA of EC-patients and tissue of the control group were incubated with the polyclonal rabbit anti-human MDC (CCL22) antibody (500-P107 1:300, Peprotech) using ZytoChem Plus HRP Polymer System mouse/rabbit following the manufacturer's instructions.

Techniques: Isolation, Immunofluorescence, Staining, Expressing

Journal: Translational Oncology

Article Title: CCL22 as an independent prognostic factor in endometrial cancer patients

doi: 10.1016/j.tranon.2024.102116

Figure Lengend Snippet: High CCL22 secretion by freshly isolated PBMCs in contrast to EC cell lines without stimulation (A). Significantly increased CCL22 in SN after coculture of PBMCs and EC cell lines (B). The addition of tumor-SN to PBMCs led to a significant increase of CCL22 levels (C), while incubation of tumor cells with PBMC-SN resulted in a decrease in CCL22 levels (D). mRNA levels of tumor cells after coculture revealed a significant increase in CCL22 of Ishikawa+ with PBMCs (E). CCL22 levels of tumor RIPA lysates also revealed a significant increase after coculture (F). All experiments were carried out in technical triplicate and repeated three times with PBMCs from different blood donors.

Article Snippet: Paraffin-embedded TMA of EC-patients and tissue of the control group were incubated with the polyclonal rabbit anti-human MDC (CCL22) antibody (500-P107 1:300, Peprotech) using ZytoChem Plus HRP Polymer System mouse/rabbit following the manufacturer's instructions.

Techniques: Isolation, Incubation