anti-il-27  (R&D Systems)

Journal: Journal of Neuroinflammation

Article Title: IL-27, a metabolic regulator secreted by astrocytes in response to GLP-1RA OHP2, modulates microglial reprogramming in Alzheimer’s disease by regulating cGAS lactylation

doi: 10.1186/s12974-025-03683-1

Figure Lengend Snippet: Blocking IL-27 attenuates both metabolic changes and the therapeutic effect of OHP2 on AD. A Schematic illustrating the design for evaluating the efficacy of Anti-IL-27 p28 in Aβ induced mice. B - C Representative images of trajectory heatmap ( B ) and recognition index ( C ) in NOR test (n=8 mice per group). D Alternation behavior in the Y-maze test in Control, Aβ 1-42 , Aβ 1-42 +OHP2, Aβ 1-42 +OHP2+anti-IL-27 p28 groups mice (n=8 mice per group). E - G The levels of IL-6, IL-1β and TNF-ɑ of mice (n=3 mice per group) were quantified using ELISA. H - I The mRNA levels of the M1 and M2 specific markers in the Control, Aβ 1-42 , Aβ 1-42 +OHP2, Aβ 1-42 +OHP2+anti-IL-27 p28 groups of microglia in astrocyte-microglia co-culture system (n=3). J - L Representative images ( J ) and quantification ( K & L ) of the level of TNF-ɑ and IL-1β as detected by WB in the Control, Aβ 1-42 , Aβ 1-42 +OHP2, Aβ 1-42 +OHP2+anti-IL-27 p28 groups (n=3). M Representative images of the phagocytic and degradation capabilities of microglia towards Aβ, as determined by Western blotting, in the Control, Aβ 1-42 , Aβ 1-42 +OHP2, Aβ 1-42 +OHP2+anti-IL-27 p28 groups. N The mRNA levels of the key metabolic enzymes Hk2, Ldha, Pkm, Pfk in in the Control, Aβ 1-42 , Aβ 1-42 +OHP2, Aβ 1-42 +OHP2+anti-IL-27 p28 groups (n=3). O - Q The glycolytic stress of primary microglia in astrocyte-microglia co-culture system following Aβ 1-42 / OHP2/ anti-IL-27 p28 treatment for 24h was evaluated using the Seahorse XFe96 system, with measurement of the extracellular acidification rate curve ( O ), as well as quantification of glycolysis ( P ) and glycolytic capacity ( Q ) (n=3 per group). The "Control", "Aβ", and "Aβ+OHP2" groups were all treated with an equivalent amount of nonspecific IgG as the vehicle control for the antibody. Data are mean ± SEM, * p<0.05, ** p<0.01, and *** p<0.001 vs Model group, # p<0.05, ## p<0.01 and ### p<0.001 vs OHP2 group. One-way ANOVA, followed by Tukey’s multiple comparisons test ( C - I , K , L , N , P & Q )

Article Snippet: The antibodies used are as follows: Rabbit monoclonal anti-Aβ (803015, 6E10, Biolegend), Rabbit monoclonal anti-TNF-α (A24214, Abclonal), Rabbit monoclonal anti-IL-1β (A22257, Abclonal), Rabbit monoclonal anti-ACTB (AC026, Abclonal), Rabbit monoclonal anti-IL-27 (sc-390482, Santa cruz, United States), Rabbit monoclonal anti-Pan-Kla (PTM-1410RM, PTM BIO, China), Rabbit monoclonal anti-cGAS (A8335, Abclonal), Rabbit monoclonal anti-p-TBK (AP1418, Abclonal), Rabbit monoclonal anti-TBK (A3458, Abclonal), Rabbit monoclonal anti-p-IRF3 (AP0623, Abclonal), Rabbit monoclonal anti-IRF3 (A2172, Abclonal), Rabbit monoclonal anti-p-mTORC1 (5536T, Cell Signaling Technology, United States), Rabbit monoclonal anti-mTORC1 (2983T, Cell Signaling Technology), Rabbit monoclonal anti-p-IκBα (82349-1-RR, Proteintech, China), Rabbit monoclonal anti-IκBα (80019-1-RR, Proteintech), Rabbit monoclonal anti-NFκB (80979-1-RR, Proteintech).

Techniques: Blocking Assay, Control, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Western Blot

Journal: Journal of Neuroinflammation

Article Title: IL-27, a metabolic regulator secreted by astrocytes in response to GLP-1RA OHP2, modulates microglial reprogramming in Alzheimer’s disease by regulating cGAS lactylation

doi: 10.1186/s12974-025-03683-1

Figure Lengend Snippet: IL-27 and OHP2 exert their effects via the glycolysis/cGAS lactylation clock/mTOR pathway. A The L-lactate levels in BV2 cells at 0, 2, 4, 8, 16, and 24 hours after the administration of Aβ 1-42 and IL-27 (n=3). B - C Representative images ( B ) and quantitative analysis ( C ) of the Pan-Kla levels in BV2 cells at 0, 2, 4, 8, 16, and 24 hours after the administration of Aβ 1-42 and IL-27 (n=3). D - E Representative images ( D ) and quantitative analysis ( E ) of the cGAS-Kla levels in BV2 cells at 0, 2, 4, 8, 16, and 24 hours after the administration of Aβ 1-42 and IL-27 (n=3). F The levels of P-TBK, TBK, P-IRF3, IRF3, P-mTORC1, mTORC1, P-IκBα, IκBα, and NF-κB in BV2 cells were measured at 4 hours after the administration of Aβ 1-42 and IL-27 (n=3). G The levels of P-TBK, TBK, P-IRF3, IRF3, P-mTORC1, mTORC1, P-IκBα, IκBα, and NF-κB in microglia of co-culture system were measured at 4 hours after the administration of Aβ 1-42 , OHP2 and anti-IL-27 p28 (n=3). H Schematic diagram of the cGAS lactylation clock pathway. Data are mean ± SEM, * p<0.05, ** p<0.01, and *** p<0.001 vs Model group, ##p<0.01 and ###p<0.001 vs the OHP2 group at the same time point. Two-way ANOVA, followed by Tukey’s multiple comparisons test ( A , C & E )

Article Snippet: The antibodies used are as follows: Rabbit monoclonal anti-Aβ (803015, 6E10, Biolegend), Rabbit monoclonal anti-TNF-α (A24214, Abclonal), Rabbit monoclonal anti-IL-1β (A22257, Abclonal), Rabbit monoclonal anti-ACTB (AC026, Abclonal), Rabbit monoclonal anti-IL-27 (sc-390482, Santa cruz, United States), Rabbit monoclonal anti-Pan-Kla (PTM-1410RM, PTM BIO, China), Rabbit monoclonal anti-cGAS (A8335, Abclonal), Rabbit monoclonal anti-p-TBK (AP1418, Abclonal), Rabbit monoclonal anti-TBK (A3458, Abclonal), Rabbit monoclonal anti-p-IRF3 (AP0623, Abclonal), Rabbit monoclonal anti-IRF3 (A2172, Abclonal), Rabbit monoclonal anti-p-mTORC1 (5536T, Cell Signaling Technology, United States), Rabbit monoclonal anti-mTORC1 (2983T, Cell Signaling Technology), Rabbit monoclonal anti-p-IκBα (82349-1-RR, Proteintech, China), Rabbit monoclonal anti-IκBα (80019-1-RR, Proteintech), Rabbit monoclonal anti-NFκB (80979-1-RR, Proteintech).

Techniques: Co-Culture Assay

Journal: Nature Communications

Article Title: Interleukin-27 is antiviral against Zika virus at the maternal-fetal interface

doi: 10.1038/s41467-025-67378-0

Figure Lengend Snippet: A Timeline for mating and treatment of human STAT2 knock-in (hSTAT2 KI) mice with high molecular weight polyinosinic-polycytidylic acid (HMW poly(I:C)). Created in BioRender. Jurado, K. (2025) https://BioRender.com/eegilr9 . B Serum IL-6 levels in PBS- and HMW Poly(I:C)-treated dams at 6 h post-injection as determined via ELISA. n = 3 dams per group. Statistical analysis performed via ordinary two-way ANOVA, **p < 0.01, ns= not significant (From left, p = >0.9999, 0.0013, 0.0011, 0.9896). Data presented as mean values ± SD. C Proportion of fetuses exhibiting healthy, early resorption, or total resorption phenotypes at E13.5 in PBS- and HMW Poly(I:C)-treated dams. Graph displays total number of fetuses from 3-4 litters per condition. D Representative images depicting fetal outcomes at E13.5. Phenotypes were determined for each fetus based on gross morphology and tissue integrity, as described in methods. E Timeline for mating and infection of hSTAT2 KI mice, as described in methods. Created in BioRender. Jurado, K. (2025) https://BioRender.com/m1xcoc2 . F Proportion of fetuses exhibiting healthy, early resorption, or total resorption phenotypes at E13.5 in PBS- and ZIKV-infected dams. Fetal outcomes were evaluated in 3 independent litters for all conditions except ZIKV-infected IL27RA −/− dams, for which 4 independent litters were evaluated. Graph displays total number of fetuses per condition. G–I ZIKV burdens at E13.5 as determined via quantitative RT-PCR. G Combined ZIKV burden of one matching fetus and placenta, normalized to combined tissue weight. Shape of data point represents observed fetal phenotype. Total number of fetal/placental units per condition plotted (see 4 F ). Statistical analysis performed via Kruskal-Wallis ANOVA, ***p < 0.001, ns=not significant (From left, p = >0.9999, 0.0006). H Left: Fetal ZIKV burdens, normalized to fetus weights. Viral burdens for subset of fetuses exhibiting “healthy” phenotypes plotted (Isotype n = 24, α-IL-27 n = 16, IL27RA −/− n = 28). Right: Placental ZIKV burdens, normalized to placental weights. Viral burdens of placentas matched to fetuses with “healthy” phenotypes plotted (Isotype n = 24, α-IL-27 n = 16, IL27RA −/− n = 28). Statistical analysis performed via Kruskal-Wallis ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns=not significant (Fetal from left, p = >0.9999, 0.0374; Placental from left, p = 0.0378, <0.0001). I Maternal serum ZIKV burdens, normalized to serum weight. n = 3 dams for Isotype and α-IL-27-treated groups, n = 4 IL27RA −/− dams. Not significant via Kruskal-Wallis ANOVA.

Article Snippet: Beginning at embryonic day 3.5 (E3.5), pregnant hSTAT2 KI dams were administered 500μg of α-IL-27 (BioXCell, BE0326, clone MM27.7B1) or isotype control (BioXCell, BE0085, clone C1.18.4) antibody via intraperitoneal injection, with subsequent treatments every three days (E6.5, E9.5, and E12.5).

Techniques: Knock-In, High Molecular Weight, Injection, Enzyme-linked Immunosorbent Assay, Infection, Quantitative RT-PCR