Human IL-22 APC-conjugated Antibody

Detection of IL‑22 in Human PBMCs stimulated to induce Th17 Cells by Flow Cytometry. CD4⁺Human peripheral blood mononuclear cells (PBMCs) either (A) stimulated to induce Th17 cells with 10 ug/mL Mouse Anti-Human CD3e Monoclonal Antibody (Catalog # MAB100), 5 ug/mL Mouse Anti-Human CD28 Monoclonal Antibody (Catalog # MAB342), 10 ng/mL Recombinant Human TGF-beta 1 (Catalog # 240-B), 20 ng/mL Recombinant Human IL-2 (Catalog # 202-IL), 20 ng/mL Recombinant Human IL-23 (Catalog # 1290-IL), 40 ng/mL Recombinant Human IL-6 (Catalog # 206-IL), and 10 ng/mL Recombinant Human IL-1 beta /IL-1F2 (Catalog # 201-LB) for 5 to 7 days, then re-stimulated with 50 ng/mL PMA and 200 ng/mL Calcium ionomycin for 2 to 4 hours or (B) resting CD4⁺PBMCs were stained with Mouse Anti-Human CD4 PE-conjugated Monoclonal Antibody (Catalog # FAB3791P) and Mouse Anti-Human IL-22 APC-conjugated Monoclonal Antibody (Catalog # IC7821A). Quadrant markers were set based on isotype control (Catalog # IC002A). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin. View our protocol for Staining Intracellular Molecules.

Detection of Human IL-22 by Flow Cytometry Frequency of IL-17 and/or IL-22 expressing mucosal CD4+T cells decreases by progression of Fiebig stage.To quantify the expression of Il-17 and IL-22 in CD4+T cells, mucosal and peripheral mononuclear cells were stimulated for 5 hours with 40 ng/mL PMA and 1 µM Ionomycin. Gating strategy of sigmoid colon Th17 and Th22 CD4+T cells is shown for a FI subject (a) and representative flow cytometry plots including unstimulated controls (unstim) gated on CD4+T cells showing expression of IL-17 and/or IL-22 in FI (b), FIII (c) and FV (d) in the sigmoid colon. A decrease in the frequency of IL-17 (e), IL-22 (f), IL-17/IL-22 (g)-producing cells was seen as well as in the subpopulation of triple-cytokine producing (IL-2, IL-22, IFN gamma ) Th17 cells (h) from FI (black circle)/FII (red circle) to FIII and FIV (red square)/FV (black square). Frequency of single and double cytokine producing cells is calculated from percentage CD4+T cells, while triple-cytokine producing cells are calculated from percentage of CD4+IL17+T cells. All comparisons were made to FI/II: *p≤0.05, **p≤0.01 and ***p≤0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25503054), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human IL-22 by Flow Cytometry Frequency of IL-17 and/or IL-22 expressing mucosal CD4+T cells decreases by progression of Fiebig stage.To quantify the expression of Il-17 and IL-22 in CD4+T cells, mucosal and peripheral mononuclear cells were stimulated for 5 hours with 40 ng/mL PMA and 1 µM Ionomycin. Gating strategy of sigmoid colon Th17 and Th22 CD4+T cells is shown for a FI subject (a) and representative flow cytometry plots including unstimulated controls (unstim) gated on CD4+T cells showing expression of IL-17 and/or IL-22 in FI (b), FIII (c) and FV (d) in the sigmoid colon. A decrease in the frequency of IL-17 (e), IL-22 (f), IL-17/IL-22 (g)-producing cells was seen as well as in the subpopulation of triple-cytokine producing (IL-2, IL-22, IFN gamma ) Th17 cells (h) from FI (black circle)/FII (red circle) to FIII and FIV (red square)/FV (black square). Frequency of single and double cytokine producing cells is calculated from percentage CD4+T cells, while triple-cytokine producing cells are calculated from percentage of CD4+IL17+T cells. All comparisons were made to FI/II: *p≤0.05, **p≤0.01 and ***p≤0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25503054), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human IL-22 by Flow Cytometry Frequency of IL-17 and/or IL-22 expressing mucosal CD4+T cells decreases by progression of Fiebig stage.To quantify the expression of Il-17 and IL-22 in CD4+T cells, mucosal and peripheral mononuclear cells were stimulated for 5 hours with 40 ng/mL PMA and 1 µM Ionomycin. Gating strategy of sigmoid colon Th17 and Th22 CD4+T cells is shown for a FI subject (a) and representative flow cytometry plots including unstimulated controls (unstim) gated on CD4+T cells showing expression of IL-17 and/or IL-22 in FI (b), FIII (c) and FV (d) in the sigmoid colon. A decrease in the frequency of IL-17 (e), IL-22 (f), IL-17/IL-22 (g)-producing cells was seen as well as in the subpopulation of triple-cytokine producing (IL-2, IL-22, IFN gamma ) Th17 cells (h) from FI (black circle)/FII (red circle) to FIII and FIV (red square)/FV (black square). Frequency of single and double cytokine producing cells is calculated from percentage CD4+T cells, while triple-cytokine producing cells are calculated from percentage of CD4+IL17+T cells. All comparisons were made to FI/II: *p≤0.05, **p≤0.01 and ***p≤0.001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25503054), licensed under a CC-BY license. Not internally tested by R&D Systems.