Journal: Clinical and Experimental Immunology

Article Title: Interleukin (IL)-22 receptor 1 is over-expressed in primary Sjogren’s syndrome and Sjögren-associated non-Hodgkin lymphomas and is regulated by IL-18

doi: 10.1111/cei.12643

Figure Lengend Snippet: List of primers for reverse transcription–polymerase chain reaction (RT–PCR) and primary and secondary antibodies used

Article Snippet: Final values were expressed as fold of induction (FOI). table ft1 table-wrap mode="anchored" t5 caption a7 GAPDH Hs99999905_m1 IL-18 Hs01038788_m1 IL-22 Hs01574154_m1 IL-22R1 Hs00222035_m1 IL-22BP Hs00364814_m1 IL-17 Hs00174383_m1 IL-23p19 Hs00372324_m1 RORc Hs01076122_m1 STAT-3 Hs00234174_m1 Rabbit anti-human IL-22 (1 : 100 dilution) Novus Biological, Littleton, CO, USA Rabbit anti-human IL-22R1 (1 : 100 dilution) Novus Biological, Littleton, CO, USA Rabbit anti-human IL-18 (1 : 100 dilution) Novus Biological, Littleton, CO, USA Rabit anti-human IL-22BP (1 : 100 dilution) Novus Biological, Littleton, CO, USA Mouse anti-human CD3 (1 : 100 dilution) Dako, Glostrup, Denmark Mouse anti-human CD68 (1 : 100 dilution) Dako, Glostrup, Denmark Mouse anti-human CD19 (1 : 100 dilution) Dako, Glostrup, Denmark Alexa Fluor® 488 Goat Anti-Rabbit IgG Life Technologies, Europe, BV, Stockholm, Sweden Alexa Fluor® 555 Goat Anti-Mouse IgG Life Technologies, Europe, BV, Stockholm, Sweden Anti-human-CD3-APC-conjugated BD Biosciences, San Jose, CA, USA Anti-human-CD19-FITC-conjugated BD Biosciences, San Jose, CA, USA Anti-human-CD14-FITC-conjugated BD Biosciences, San Jose, CA, USA Anti-human-CD68-FITC-conjugated BD Biosciences, San Jose, CA, USA Anti-human-IL-22R1-PE-conjugated R&D Systems, Minneapolis, MN, USA Anti-human-CD4-APC-conjugated BD Biosciences, San Jose, CA, USA Anti-human-IL-17-FITC-conjugated R&D Systems, Minneapolis, MN, USA Open in a separate window APC = allophycocyanin; FITC = fluorescein isothiocyanate; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; Ig = immunoglobulin; PE = phycoerythrin; STAT-3 = signal transducer and activator of transcription-3.

Techniques: Reverse Transcription Polymerase Chain Reaction

Journal: iScience

Article Title: Peripheral immunophenotyping of AITD subjects reveals alterations in immune cells in pediatric vs adult-onset AITD

doi: 10.1016/j.isci.2021.103626

Figure Lengend Snippet: Manual gating analysis of myeloid and NK cell subsets reveals pediatric AITD patients show a tendency of reduced frequencies of total NK and the cytotoxic CD56 lo NK subset compared to pediatric controls (A) Frequencies of dendritic cell (DC) subsets are similar among AITD and control subjects. Conventional type 1 DCs (cDC1) were gated as CD3-CD19-HLA-DR + CD14-CD16-CD11c + CD1c-, conventional type 2 DCs (cDC2) were gated as CD3-CD19-HLA-DR + CD14-CD16-CD11c + CD1c+, and plasmacytoid DC (pDC) were gated as CD3-CD19-HLA-DR + CD45RA + CD123 + CD11c-. (B) The frequency of classical monocytes (CD3-CD19-HLA-DR + CD14+) is increased in pediatric AITD subjects compared to pediatric controls, whereas the frequency of intermediate monocytes (CD3-CD19-HLA-DR + CD14 + CD16+) and non-classical monocytes (CD3-CD19-HLA-DR + CD14-CD16+) are similar among AITD and control subjects. (C) Frequencies of cytokine producing monocytes at baseline demonstrates adult AITD patients have reduced frequencies of IL-23p19+ and IL-6+ monocytes compared to adult controls. (D) The frequency of natural killer (NK; CD3-CD19-HLA-DR-CD16+) cells is reduced, but not significantly, in pediatric AITD and adult AITD patients compared to controls. The reduced frequency of NK cells appears relegated to the cytotoxic CD56 lo NK cell subset, as opposed to the regulatory CD56 hi NK subset. PA: pediatric AITD, PH: pediatric healthy control, AA: adult AITD, AH: adult healthy control. Data are represented as mean ± SEM. Samples were processed as depicted in Figure S1 . Frequency of myeloid and NK cell subsets, and cytokine production were determined by manual gating, as depicted in and . Statistical significance determined by One-way ANOVA with Sidak multiple comparisons post-test. ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Mouse monoclonal anti-human IL-23p19 (clone 23dcdp) , eBioscience , Cat#12-7823-42 RRID: AB_10668835.

Techniques: Control

Journal: iScience

Article Title: Peripheral immunophenotyping of AITD subjects reveals alterations in immune cells in pediatric vs adult-onset AITD

doi: 10.1016/j.isci.2021.103626

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-human IL-23p19 (clone 23dcdp) , eBioscience , Cat#12-7823-42 RRID: AB_10668835.

Techniques: Recombinant, Antibody Labeling, Immunopeptidomics, Mass Cytometry, Software

Journal: Scientific Reports

Article Title: Chemotherapy-induced high expression of IL23A enhances efficacy of anti-PD-1 therapy in TNBC by co-activating the PI3K-AKT signaling pathway of CTLs

doi: 10.1038/s41598-024-65129-7

Figure Lengend Snippet: IL23A was significantly upregulated by chemotherapy and associated with a favorable prognosis in TNBC patients. ( A ) The whole transcriptomic sequencing of TNBC cells treated with chemotherapeutic drugs. Upper panel, the experimental design. Lower panel, overlapping immune- and secretome-related genes, and significantly affected genes in TNBC cells treated with chemotherapeutic drugs (log2|Fc|≥ 2, P < 0.05). ( B ) Heapmap of 18 genes identified in Fig. 1A, ranked by normalized log2Fc. IL23A was identified as the most upregulated gene in response to chemotherapeutic drugs. ( C ) mRNA levels of two subunits of IL-23 (IL23A and IL12B) in TNBC cells treated with chemotherapeutic drugs, detected by qRT-PCR. ( D ) Protein expressions and quantifications of IL-23 in TNBC cells treated with chemotherapeutic drugs, detected by western blot. The PVDF membranes were cropped at 35 kDa before antibody incubation. Original blots are presented in Supplementary Fig. . ( E ) Left panel, representative immunohistochemistry (IHC) staining of IL-23A and CD8 on resected tumor tissues from TNBC patients who did (n = 27) or did not (n = 37) receive neoadjuvant chemotherapy. Scale bars 50 μm. Middle and right panel, quantitative analysis. ( F ) Spearman correlation analysis revealing a correlation between IL-23A staining score and CD8 + T cell counts in resected tissues from TNBC patients (n = 64, r = 0.465, p = 0.0001). ( G ) Upper panel, representative CT scans before and after neoadjuvant chemotherapy from one of the patients whose resected tumor tissues exhibited high scores of IL-23A staining, as well as the representative IHC staining. Scale bars, 50 μm. Lower panel, the slope chart shows the overall response rate (ORR) of TNBC patients who received neoadjuvant chemotherapy (n = 22). Blue bars represent patients from the IL-23A low group with an IL-23A IHC score of 0–6 (n = 8); Red bars represent patients from the IL-23A high group with an IL-23A IHC score of 7–12 (n = 14). (H) The correlation between IL23A mRNA levels and overall survival of patients with breast cancer (HR = 0.73, P = 0.0048, left panel) or Triple-negative breast cancer (HR = 0.22, P < 0.0001, right panel). Data from the Kaplan–Meier plotter database ( https://www.kmplot.com/ ). Error bars represent standard deviation (SD), *P < 0.05; ** P < 0.01; ****P < 0.0001.

Article Snippet: After blocking with 1% BSA (Beyotime, China), the slides were incubated with primary antibodies [IL-23A (1:500; Proteintech, China), CD8α (1:200; Abcam, UK)] overnight at 4 °C and then the corresponding secondary antibody (ZSGB-BIO, China) for 30 min at room temperature.

Techniques: Sequencing, Quantitative RT-PCR, Western Blot, Incubation, Immunohistochemistry, Staining, Standard Deviation

Journal: Scientific Reports

Article Title: Chemotherapy-induced high expression of IL23A enhances efficacy of anti-PD-1 therapy in TNBC by co-activating the PI3K-AKT signaling pathway of CTLs

doi: 10.1038/s41598-024-65129-7

Figure Lengend Snippet: Clinical parameters of patients for response evaluation in this study.

Article Snippet: After blocking with 1% BSA (Beyotime, China), the slides were incubated with primary antibodies [IL-23A (1:500; Proteintech, China), CD8α (1:200; Abcam, UK)] overnight at 4 °C and then the corresponding secondary antibody (ZSGB-BIO, China) for 30 min at room temperature.

Techniques: Significance Assay

Journal: Heliyon

Article Title: IL-23 production in human macrophages is regulated negatively by tumor necrosis factor α-induced protein 3 and positively by specificity protein 1 after stimulation of the toll-like receptor 7/8 signaling pathway

doi: 10.1016/j.heliyon.2022.e08887

Figure Lengend Snippet: Effect of Lipofectamine on IL-23 expression. IL-23p19 mRNA levels were determined in macrophages treated with or without Lipofectamine by real-time reverse transcription polymerase chain reaction (RT-PCR) (a). p53 protein levels in macrophages treated with or without Lipofectamine for 6 h were measured by Western blotting. A representative blot is shown. Data were obtained by using samples from 3 individuals in each group. Results are shown as the mean (SE). ∗∗ P < .01 (with Bonferroni correction) (b). Furthermore, IL-23 protein levels in macrophages transfected with siRNA for p53, TNFα, IRF5, TRAF6, OAS-1, or cryopyrin after exposure to resiquimod were determined by enzyme-linked immunosorbent assay (c and d). Data were obtained by using samples from 3 individuals in each group. Results are shown as the mean (SE). ∗∗ P < .01; ∗ P < .05 (with Bonferroni correction); N.S., not significant IL-23p19 protein levels were measured by Western blotting in macrophages transfected with siRNA for IRF5, p53, TNFα, OAS-1 or cryopyrin after stimulation with resiquimod. A representative blot is shown. Data were obtained by using samples from 3 individuals in each group. Results are shown as the mean (SE). ∗ P < .05 (with Bonferroni correction); N.S., not significant (e). Furthermore, OAS-1 protein levels were measured by Western blotting in macrophages treated with or without Lipofectamine. A representative blot is shown. Data were obtained by using samples from 3 individuals in each group. Results are shown as the mean (SE). ∗ P < .05 (with Bonferroni correction); N.S., not significant (f).

Article Snippet: Western blotting for IL-23, TNFAIP3, Sp1, TGFβ1, cyclooxygenase-2 and OAS-1 After macrophages were exposed to LPS (10 ng/mL), or resiquimod (5μM) for 6 h, production of the proteins for IL-23p19, TNFAIP3, Sp1, TGFβ1, and cyclooxygenase-2 (COX-2) were assessed by Western blotting with the following antibodies: IL-23p19 (catalog#sc-271279, Santa Cruz Biotechnology, Santa Cruz, CA), TNFAIP3 catalog#sc-166692, Santa Cruz Biotechnology), Sp1 (catalog#sc-17824, Santa Cruz Biotechnology), TGFβ1 (catalog#sc-130348, Santa Cruz Biotechnology), and COX-2 (catalog#sc-376861, Santa Cruz Biotechnology).

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Aspergillus fumigatus-induced IL-22 is not restricted to a specific Th cell subset and is dependent on complement receptor 3.

doi: 10.4049/jimmunol.1202601

Figure Lengend Snippet: FIGURE 5. Role of IL-23 and IL-1 in A. fumigatus–induced IL-17A and IL-22. (A) IL-1b and IL-23 concentrations in PBMCs (2.5 3 106/ml) (n = 7) that were stimulated with HI A. fumigatus conidia or hyphae at 37C and 5% CO2 for 24 h. IL-17 and IL-22 were measured in the culture supernatants of PBMCs (2.5 3 106/ml) stimulated with HI A. fumigatus conidia in the presence or absence of 10 mg/ml IL-1Ra (B) (n = 7 for IL-17, n = 12 for IL-22), IL-23 (C) (n = 12), or anti–IL-23p19 (D) (n = 5 for IL-17, n = 6 for IL-22). Differences of the means were analyzed for significance using the Wilcoxon signed rank test. *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: IL-1R signaling was blocked by its natural receptor antagonist (Ra) IL-1Ra (10 mg/ml) (Amgen, Thousand Oaks, CA), and IL-23 was blocked with mouse anti-human IL-23p19 (10 mg/ml) (R&D Systems).

Techniques: