Journal: iScience

Article Title: Interferon-γ and IL-27 positively regulate type 1 regulatory T cell development during adaptive tolerance

doi: 10.1016/j.isci.2025.112308

Figure Lengend Snippet:

Article Snippet: Mouse Anti-Mouse IL-27 p28 (clone MM27.7B1) , BioXCell , Cat #BE0326; RRID:AB_2819053.

Techniques: Recombinant, Saline, Staining, Red Blood Cell Lysis, Sequencing, Software

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

Article Title: Suppression of ongoing experimental autoimmune encephalomyelitis by neutralizing the function of the p28 subunit of IL-27.

doi: 10.4049/jimmunol.173.10.6465

Figure Lengend Snippet: FIGURE 1. DNA vaccination-based anti-IL-27 Abs are highly specific. The Western blot shows that our DNA vaccination-based anti-IL-27 Ab binds mouse IL-27 p28 (lane 1; 27 kDa), but not recombinant mouse IL-18, IL-12, or TNF- (lanes 2, 3, and 4, respectively). A, Coomassie Blue staining verifies the appearance of each cytokine on the loaded gel. B, Western blot showing that of these cytokines, our DNA vaccination-based anti-IL-27 Ab binds only mouse IL-27 p28. These Ab also bound natural mouse IL-27 (verified by sequencing) from supernatant of activated MOGp35–55-specific cultured primary draining lymph node cells (not shown).

Article Snippet: Our recombinant mouse IL-27 p28, produced as described above, and commercially available recombinant mouse IL-18, IL-12, and TNF- (R&D Systems) were each subjected to Western blot analysis according to the protocol described in detail previously (29, 32), with the minor modification of using a 12% (rather than 8%) running gel.

Techniques: Western Blot, Recombinant, Staining, Sequencing, Cell Culture

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

Article Title: Suppression of ongoing experimental autoimmune encephalomyelitis by neutralizing the function of the p28 subunit of IL-27.

doi: 10.4049/jimmunol.173.10.6465

Figure Lengend Snippet: FIGURE 2. Anti-p28 Abs suppresses ongoing severe EAE. A, Four groups of 10 mice each were subjected to MOGp35–55-induced EAE. Beginning at the onset of disease (day 17), these mice were repeatedly (every other day) administered 100 g/mouse of anti-p28 Ab (f), IgG obtained from naive Lewis rats (Œ), or PBS (E). An observer blind to the experimental procedure scored EAE daily. The experiment summarized in Fig. 2 shows the results of one of three experiments performed under similar experimental conditions, with similar results. The mean maximal score SE represents six mice per group. The other four mice were killed on day 30 and subjected to histological evaluation (see Fig. 3). B, Five groups of six mice each were subjected to MOGp35–55-induced EAE. Beginning at the onset of disease (day 17), these mice were repeatedly (every other day) administered 100 g of anti-p28 Ab/mouse (f), anti-IL-18 Ab (F), anti-IL-1 Ab (), IgG obtained from Lewis rats previously subjected to an empty plasmid administration (Œ), or PBS (E). An observer blind to the experimental procedure scored EAE daily. Results are shown as the mean maximal score SE of six mice per group. C, Three groups of six mice each were subjected to induction of transferred EAE. Beginning at the onset of disease (day 5), these mice were repeatedly (every other day) administered 100 g of anti-p28 Ab/mouse (f), IgG obtained from naive Lewis rats (Œ), or PBS (E). An observer blind to the experimental procedure scored EAE daily. Results are shown as mean maximal score SE of six mice per group.

Article Snippet: Our recombinant mouse IL-27 p28, produced as described above, and commercially available recombinant mouse IL-18, IL-12, and TNF- (R&D Systems) were each subjected to Western blot analysis according to the protocol described in detail previously (29, 32), with the minor modification of using a 12% (rather than 8%) running gel.

Techniques: Plasmid Preparation

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

Article Title: Suppression of ongoing experimental autoimmune encephalomyelitis by neutralizing the function of the p28 subunit of IL-27.

doi: 10.4049/jimmunol.173.10.6465

Figure Lengend Snippet: FIGURE 4. The beneficial effect of anti-IL-27 is dependent on the con- tinuing administration of protective Abs. Three groups of six mice each were subjected to active induction of EAE. Beginning 1 day after the onset of disease (day 17), these mice were treated with either a single dose of anti-p28 Ab (100 g/mouse; E) or with repeated administration (every other day) of this Ab (Œ) or PBS (f). An observer blind to the experi- mental procedure scored EAE daily. Results are shown as the mean max- imal score SE of six mice per group.

Article Snippet: Our recombinant mouse IL-27 p28, produced as described above, and commercially available recombinant mouse IL-18, IL-12, and TNF- (R&D Systems) were each subjected to Western blot analysis according to the protocol described in detail previously (29, 32), with the minor modification of using a 12% (rather than 8%) running gel.

Techniques:

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

Article Title: Suppression of ongoing experimental autoimmune encephalomyelitis by neutralizing the function of the p28 subunit of IL-27.

doi: 10.4049/jimmunol.173.10.6465

Figure Lengend Snippet: FIGURE 3. Anti-IL-27 therapy reduces the histological score of EAE. Histological evaluation was conducted 30 days after disease induction. Lumbar spinal cord samples from naive mice or from EAE mice treated with PBS, IgG from naive mice, or anti-IL-27 p28 Abs were subjected to histological analysis (nine sections each group). The arrowheads point to the parenchymal mononuclear cell infiltration. The scale for mononuclear cell infiltration used was: 0, no mononuclear cell infiltration; 1, one to five perivascular lesions per section with minimal parenchymal infiltration; 2, five to 10 perivascular lesions per section with parenchymal infiltration; and 3, 10 perivascular lesions per section with extensive parenchymal infiltration. The mean histological score SE was calculated for each group.

Article Snippet: Our recombinant mouse IL-27 p28, produced as described above, and commercially available recombinant mouse IL-18, IL-12, and TNF- (R&D Systems) were each subjected to Western blot analysis according to the protocol described in detail previously (29, 32), with the minor modification of using a 12% (rather than 8%) running gel.

Techniques:

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

Article Title: Suppression of ongoing experimental autoimmune encephalomyelitis by neutralizing the function of the p28 subunit of IL-27.

doi: 10.4049/jimmunol.173.10.6465

Figure Lengend Snippet: FIGURE 5. Protective administration of anti-IL-27 Abs decreases in vivo polarization of CD4 T cells into Th1 and suppresses IFN- production by Ag-specific T cells. C57BL/6 mice (three per group) were subjected to active induction of EAE and then to repeated administration (days 12, 14, and 16) of anti-IL-27 p28 Abs (100 g), PBS, or normal rat IgG. On day 17 cervical lymph node cells (that drain the autoimmune site) were subjected to intracellular staining of IL-4 and IFN-. A, FACS analysis of CD4 T cells in this experiment. This experiment represents results obtained in three different independent experiments with very similar data. Subsequently, cervical lymph node T cells from these mice were cultured in the presence of 100 M MOGp35–55. After 72 h of stimulation, supernatants were assayed for the protein level of IFN- (B) and IL-4 (not shown). This experiment represents results obtained in three different independent experiments with very similar data.

Article Snippet: Our recombinant mouse IL-27 p28, produced as described above, and commercially available recombinant mouse IL-18, IL-12, and TNF- (R&D Systems) were each subjected to Western blot analysis according to the protocol described in detail previously (29, 32), with the minor modification of using a 12% (rather than 8%) running gel.

Techniques: In Vivo, Staining, Cell Culture

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

Article Title: Suppression of ongoing experimental autoimmune encephalomyelitis by neutralizing the function of the p28 subunit of IL-27.

doi: 10.4049/jimmunol.173.10.6465

Figure Lengend Snippet: FIGURE 6. Neutralizing the function of IL-27 reduces IFN- produc- tion by IFN--producing T cells. A, C57BL/6 mice (three per group) were subjected to active induction of EAE and then to repeated administration (days 3 and 6) of 100 g of anti-IL-27 p28 Abs (group 3), PBS (group 2), or normal rat IgG (group 1). On day 9, spleen cells were subjected to spot ELISA as previously described (38). A, Relative number of positive spots per 107 cultured cells. The average size of positive spots was analyzed. B, The MOGp33–55-specific CD4 T cell line was cultured with or without 100 M MOGp33–55. Cultured cells were supplemented with anti-IL-27 Abs at a final concentration of 10 g/ml (), normal rat IgG (f), or PBS (E). After 60 h of incubation, cells were plates in spot ELISA plates for an additional 24 h for the detection of IFN--positive spots (38). Number of positive spots (y-axis) and spot sizes (x-axis; logarithmic scale) determined as previously described (46).

Article Snippet: Our recombinant mouse IL-27 p28, produced as described above, and commercially available recombinant mouse IL-18, IL-12, and TNF- (R&D Systems) were each subjected to Western blot analysis according to the protocol described in detail previously (29, 32), with the minor modification of using a 12% (rather than 8%) running gel.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Concentration Assay, Incubation

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: The expression of sIL-27Rα level in GVHD patients. ( A ) Serum sIL-27Rα levels were significantly increased on the day of neutrophil engraftment compared with pre-conditioning ( P < 0.0001). ( B ) The serum sIL-27Rα levels in patients with grade II–IV aGVHD were significantly lower than those of 0-I aGVHD patients on the day of neutrophil engraftment ( P < 0.001). ( C ) On the day of neutrophil engraftment, serum sIL-27Rα levels in patients with skin aGVHD ( P < 0.01) and liver aGVHD ( P < 0.05) were significantly lower than those of 0-I aGVHD patients. ( D ) There were no similar significant difference regarding cGVHD. Data shown are mean ± SD.

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: The diagnostic value of sIL-27Rα in aGVHD and the association of sIL-27Rα with aGVHD severity, relapse and survival. ( A ) The area under the ROC curve (AUC) was 0.735 (95% CI 0.618–0.853, P = 0.001) on the day of neutrophil engraftment; when using 59.4 ng/ml as cut-off value from the ROC curve, the sensitivity and specificity were 56% and 81%, respectively. ( B ) The cumulative incidence of grade II–IV aGVHD was significantly lower in patients with high sIL-27Rα levels by Gray’s test ( P = 0.004). ( C ) Patients with high sIL-27Rα levels showed favourable overall survival compared with patients with low sIL-27Rα levels with Kaplan-Meier survival analysis by log rank test ( P < 0.001). ( D , E ) Patients with high sIL-27Rα levels had lower relapse rate (CIR) and non-relapse mortality (NRM) than did patients with low sIL-27Rα levels by Gray’s test (P = 0.008, respectively).

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: Diagnostic Assay

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: The association of sIL-27Rα levels at neutrophil engraftment with clinical factors.

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: Biomarker Discovery

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: Univariate and multivariate analyses of factors affecting the incidence of grade II–IV acute graft- versus-host disease after allogeneic hematopoietic stem cell transplantation at neutrophil engraftment.

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: Transplantation Assay

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: sIL-27Rα as an independent prognostic aGVHD biomarker validated in an independent cohort of 80 patients. ( A ) The area under the ROC curve (AUC) was 0.790 (95% CI 0.688–0.892, P < 0.001) on the day of neutrophil engraftment, using 59.4 ng/ml as the cut-off value. ( B ) The cumulative incidence of grade II–IV aGVHD was significantly lower in patients with high sIL-27Rα levels by Gray’s test ( P < 0.001). ( C ) Patients with high sIL-27Rα levels showed favourable overall survival compared with patients with low sIL-27Rα levels on Kaplan-Meier survival analysis by log rank test ( P = 0.012). ( D , E ) Patients with high sIL-27Rα levels had similar relapse rates (CIR) and lower non-relapse mortality (NRM) than did patients with low sIL-27Rα levels by Gray’s test ( P = 0.0931, P = 0.005, respectively).

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: Biomarker Discovery

Journal: Scientific Reports

Article Title: Soluble interleukin-27 receptor alpha is a valuable prognostic biomarker for acute graft-versus-host disease after allogeneic haematopoietic stem cell transplantation

doi: 10.1038/s41598-018-28614-4

Figure Lengend Snippet: Association of sIL-27Rα with serum cytokine levels in aGVHD. The associations between sIL-27Rα levels with serum IL-27, IL-10, HGF, TNFR1, elafin, REG3α and ST2 levels were analysed by using Spearman’s rank correlation coefficient.

Article Snippet: After washing, biotinylated goat anti-human IL-27Rα polyclonal antibody (100 μL/well, Catalogue #BAF1479, R&D Systems) was added as detection antibody for 2 h at room temperature.

Techniques: