Journal: International Journal of Molecular Sciences

Article Title: Human IL-23R Cytokine-Binding Homology Region-Fc Fusion Protein Ameliorates Psoriasis via the Decrease of Systemic Th17 and ILC3 Cell Responses

doi: 10.3390/ijms20174170

Figure Lengend Snippet: Construction, expression, and purification of rhIL-23R-CHR/Fc. ( A ) PCR products of rhIL-23R-CHR/Fc recombinant gene. Lane 1, rhIL-23R-CHR/Fc gene sequence. Lane 2, hIgG1 Fc fragment. Lane 3, rhIL-23R-CHR gene. M, DNA molecular weight markers, bp. ( B ) Identification of rhIL-23R-CHR-Fc/T vector digested by Hind III and Xho I sites. Lane 1, pcDNA3.1 (+) - IL-23R-CHR/Fc after digestion. Lane 2, pcDNA3.1 (+) - IL-23R-CHR/Fc before digestion. M, DNA marker, bp. ( C ) Plasmid map of pcDNA3.1 (+) - IL-23R-CHR/Fc. The gene sequence encoding rhIL-23R-CHR/Fc was inserted into pcDNA3.1 (+) vector at the corresponding restriction sites Hind III and Xho I. ( D ) SDS-PAGE analysis of the purified rhIL-23R-CHR/Fc fusion protein using Protein A column. Lane 1, purified rhIL-23R-CHR/Fc fusion protein at reduced state. M, protein molecular weight markers, KDa. ( E ) Western blot analysis of rhIL-23R-CHR/Fc using mAbs against human IL-23R. Lane 1, purified rhIL-23R-CHR/Fc fusion protein at non-reduced state. Lane 2, purified rhIL-23R-CHR/Fc fusion protein at reduced state. ( F ) Plasma clearance of IL23RCHR-Fc in mice.

Article Snippet: The rhIL23R-CHR/Fc fragment cleaved from rhIL-23R-CHR/Fc/T Vector pMD19 (Takara, Tokyo, Japan) with Xho I and Hind III (Takara, Tokyo, Japan).

Techniques: Expressing, Purification, Recombinant, Sequencing, Molecular Weight, Plasmid Preparation, Marker, SDS Page, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Human IL-23R Cytokine-Binding Homology Region-Fc Fusion Protein Ameliorates Psoriasis via the Decrease of Systemic Th17 and ILC3 Cell Responses

doi: 10.3390/ijms20174170

Figure Lengend Snippet: Role of rhIL-23R-CHR/Fc in mouse Th17 cell differentiation. Mouse naive CD4 + T cells were differentiated into Th17 cells in the presence of different concentrations of rhIL23R-CHR/Fc in vitro. ( A ) Analysis of Th17 cells (CD4 + IL-17 + ) by flow cytometry. ( B ) The percentage of Th17 cells. ( C ) IL-17A level in supernatants derived from cultured spleen cells. All tests were performed in triplicate and are presented as the mean ± SEM. * p < 0.05; ** p < 0.01; and *** p < 0.001.

Article Snippet: The rhIL23R-CHR/Fc fragment cleaved from rhIL-23R-CHR/Fc/T Vector pMD19 (Takara, Tokyo, Japan) with Xho I and Hind III (Takara, Tokyo, Japan).

Techniques: Cell Differentiation, In Vitro, Flow Cytometry, Derivative Assay, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Human IL-23R Cytokine-Binding Homology Region-Fc Fusion Protein Ameliorates Psoriasis via the Decrease of Systemic Th17 and ILC3 Cell Responses

doi: 10.3390/ijms20174170

Figure Lengend Snippet: rhIL-23R-CHR/Fc ameliorated skin inflammation in an imiquimod (IMQ)-induced psoriasis-like model. ( A ) Schematic diagram of intravenous administration of rhIL23R-CHR/Fc on days 2, 4 and 6 during the application of IMQ. Three mice in each group were sacrificed on days 4, 7, 10 and 14 to conduct experiments. The normal group did not receive IMQ as a negative control. Cyclosporin A (CsA), an immunosuppressive agent commonly used in the clinical therapy for autoimmune diseases, was used in the treatment of psoriasis at 1.5 mg/kg as a positive control. ( B ) Phenotypic presentation of lesional skin from mice. ( C ) The size of spleens of mice. ( D ) Mice skin Psoriasis Area and Severity Index (PASI) scores. ( E ) Haematoxylin and eosin staining of lesional skin obtained from normal mice or mice treated with IMQ, CsA or rhIL-23R-CHR/Fc at day 7 (200×, 400×).

Article Snippet: The rhIL23R-CHR/Fc fragment cleaved from rhIL-23R-CHR/Fc/T Vector pMD19 (Takara, Tokyo, Japan) with Xho I and Hind III (Takara, Tokyo, Japan).

Techniques: Negative Control, Positive Control, Staining

Journal: International Journal of Molecular Sciences

Article Title: Human IL-23R Cytokine-Binding Homology Region-Fc Fusion Protein Ameliorates Psoriasis via the Decrease of Systemic Th17 and ILC3 Cell Responses

doi: 10.3390/ijms20174170

Figure Lengend Snippet: rhIL-23R-CHR/Fc inhibited Th17 cell-mediated inflammatory response. ( A ) Representative flow-cytometric analysis of Th17 cells (CD4 + IL-17A + ) in spleen from normal mice or mice ( n = 3) treated with IMQ at day 4, 7, 10, and 14. ( B ) Statistical data of Th17 cell percentage. ( C , D ) Protein levels of mIL-17A(C) and mIL-23(D) in mice serum. Data are mean ± SEM of n ≥ 3 per group. * p < 0.05, ** p < 0.01, and *** p < 0.001. ns, not significant (Student’s t -test).

Article Snippet: The rhIL23R-CHR/Fc fragment cleaved from rhIL-23R-CHR/Fc/T Vector pMD19 (Takara, Tokyo, Japan) with Xho I and Hind III (Takara, Tokyo, Japan).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Human IL-23R Cytokine-Binding Homology Region-Fc Fusion Protein Ameliorates Psoriasis via the Decrease of Systemic Th17 and ILC3 Cell Responses

doi: 10.3390/ijms20174170

Figure Lengend Snippet: rhIL-23R-CHR/Fc inhibited the transcriptional levels of inflammatory-associated cytokines, transcription factors, and chemokines in mouse skin and spleen. ( A – D ) The mRNA expression of IL-17A, IL-17F, IL-22 ( A ), Rorγt ( B ), IL-23R ( C ) and CCL20 ( D ) in skin lesions from normal or mice treated with IMQ ( n > 3). ( E , F ) The mRNA expression of IL-17A, IL-17F ( E ), and IL-23R ( F ) in spleen from normal or mice treated with IMQ. All results are representative of at least 3 independent experiments with at least 3 samples in each group. Data are mean ± SEM. * p < 0.05, ** p < 0.01 and *** p < 0.001. ns, not significant (Student’s t -test).

Article Snippet: The rhIL23R-CHR/Fc fragment cleaved from rhIL-23R-CHR/Fc/T Vector pMD19 (Takara, Tokyo, Japan) with Xho I and Hind III (Takara, Tokyo, Japan).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Human IL-23R Cytokine-Binding Homology Region-Fc Fusion Protein Ameliorates Psoriasis via the Decrease of Systemic Th17 and ILC3 Cell Responses

doi: 10.3390/ijms20174170

Figure Lengend Snippet: rhIL-23R-CHR/Fc limited the production of ILC3 cells. ( A ) Percentages of ILC3s (Lin − CD45 + RORγt + ) in small intestine lamina propria from Normal, Model, rhIL-23R-CHR/Fc, and CsA group mice were analyzed by flow cytometry ( n = 3) at day 7, 10, and 14. ( B ) Statistical data of ILC3 cells percentage. ( C ) The frequency of Nkp46 − / + ILC3 cells at day 7 from Normal, Model, rhIL-23R-CHR/Fc, and CsA group mice. ( D ) Analysis of IL-22 level in small intestines by immunofluorescence staining (200×). All results are representative of at least 3 independent experiments with at least 3 samples per group in each. Data represent the mean ± SEM. * p < 0.05 (Student’s t -test).

Article Snippet: The rhIL23R-CHR/Fc fragment cleaved from rhIL-23R-CHR/Fc/T Vector pMD19 (Takara, Tokyo, Japan) with Xho I and Hind III (Takara, Tokyo, Japan).

Techniques: Flow Cytometry, Immunofluorescence, Staining