Journal: Science Advances

Article Title: Messenger RNA delivery of a dual CD25/CD122 affinity-tuned IL-2 variant prolongs exposure and potentiates antitumor T cell immunity

doi: 10.1126/sciadv.aeh9215

Figure Lengend Snippet: ( A ) IL-2 variants evaluated in this study. ( B ) Affinity measurements of IL-2 variants to IL-2Rα and IL-2Rβ as determined by biolayer interferometry. K d , dissociation constant. ( C ) Signal transducer and activator of transcription 5 (STAT5) phosphorylation in T cell subsets within human peripheral blood mononuclear cells (PBMCs; n = 5 donors) was analyzed following stimulation with serial dilutions of human embryonic kidney (HEK) 293T supernatants containing Alb–IL-2 variants, as determined by flow cytometry. The potency of Alb–IL-2 variants is expressed as reciprocal median effective concentration (EC 50 ). For potencies below the lower limit of quantification (LLOQ), LLOQ/2 values were used for EC 50 . ( D ) STAT5 phosphorylation in IL-2Rα RNA-electroporated or nonelectroporated CD8 + T cells isolated from PBMCs ( n = 2 donors) upon incubation with HEK293T supernatants containing Alb–IL-2 variants as determined by flow cytometry. ( E ) RiboCytokine platform technology. Albumin-cytokine fusion protein sequences are encoded by modified single-stranded RNA and formulated into LNPs. ( F ) Bioluminescence in vivo imaging of BALB/c mice ( n = 3) injected intravenously with 1 μg of firefly luciferase RNA-LNP. Alb RNA-LNP served as control. h, hours. ( G ) Serum levels of translated Alb-IL–2var protein in BALB/c mice ( n = 3) injected intravenously with 10 μg of Alb–IL-2var RNA-LNP (Alb–IL-2var). Alb RNA-LNP served as the control. ( H ) Flow cytometry analysis of CD8 + and CD4 + T cells, T reg cells and CD8 + T–to–T reg cell ratio in blood of C57BL/6 mice ( n = 3) 7 days after dosing with 10 μg of Alb–IL-2 RNA-LNP (Alb–IL-2) or Alb–IL-2var RNA-LNP (Alb–IL-2var). Alb RNA-LNP served as the control. Horizontal dotted lines represent the means of the control group (H). One-way analysis of variance (ANOVA) with Dunnett’s post hoc test (C and H). Not significant (n.s.), P > 0.05; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001.

Article Snippet: Genes encoding the different IL-2 variants with an N-terminal His 6 -tag were synthesized and cloned into pTwist_CMV_BetaGlobin_WPRE_Neo expression vector (Twist Bioscience).

Techniques: Phospho-proteomics, Flow Cytometry, Concentration Assay, Isolation, Incubation, Modification, In Vivo Imaging, Injection, Luciferase, Control

Journal: Science Advances

Article Title: Messenger RNA delivery of a dual CD25/CD122 affinity-tuned IL-2 variant prolongs exposure and potentiates antitumor T cell immunity

doi: 10.1126/sciadv.aeh9215

Figure Lengend Snippet: ( A to D ) CT26 tumor-bearing BALB/c mice ( n = 11) were treated with gp70 RNA-LPX vaccine (vaccine), Alb–IL-2 RNA-LNP (Alb–IL-2), or Alb–IL-2var RNA-LNP (Alb–IL-2var), starting on day 10 post–tumor inoculation (average tumor volume, 16 mm 3 ). Alb RNA-LNP served as the control. (A) Experimental design. (B) gp70-specific CD8 + T and T reg cells, and (C) gp70-specific CD8 + T–to–T reg cell ratio in blood 7 days after the first treatment, as determined by flow cytometry. (D) Survival and individual tumor growth curves. Horizontal dotted lines represent the means of the control group (B and C), and vertical dotted lines represent treatments (D). ( E to G ) B16F10 tumor-bearing C57BL/6 mice ( n = 15) were treated with Trp-1 RNA-LPX vaccine (vaccine), Alb–IL-2 RNA-LNP (Alb–IL-2), and/or Alb–IL-2var RNA-LNP (Alb–IL-2var), starting on day 8 post–tumor inoculation (average tumor volume, 3 mm 3 ). Irrelevant RNA-LPX and hAlb RNA-LNP served as a control. (E) Experimental design. (F) Trp-1–specific CD8 + T cells 7 days after the second treatment, T reg cells 7 days after the first treatment, and CD8 + T–to–T reg cell ratio 7 days after the second treatment, in blood, as determined by flow cytometry. (G) Survival and individual tumor growth curves. Horizontal dotted lines represent the means of the control group (F), and vertical dotted lines represent treatments (G). Statistical tests: Kruskal-Wallis test with Dunn’s multiple comparison test (B, C, and F), log-rank test (D, left; and G, left), and two-way ANOVA with Dunnett’s multiple comparison test (G, right). Not significant (n.s.), P > 0.05; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001. CR, complete regression; sc, subcutaneous; Trp-1, tyrosinase-related protein 1.

Article Snippet: Genes encoding the different IL-2 variants with an N-terminal His 6 -tag were synthesized and cloned into pTwist_CMV_BetaGlobin_WPRE_Neo expression vector (Twist Bioscience).

Techniques: Control, Flow Cytometry, Comparison

Journal: Science Advances

Article Title: Messenger RNA delivery of a dual CD25/CD122 affinity-tuned IL-2 variant prolongs exposure and potentiates antitumor T cell immunity

doi: 10.1126/sciadv.aeh9215

Figure Lengend Snippet: ( A ) Serum levels of translated Alb–IL-2var in female cynomolgus monkeys ( n = 3) injected intravenously with 50 μg (mean of 19.45 μg/kg) of Alb–IL-2var RNA-LNP (Alb–IL-2var). ( B to E ) Lymphocytes as assessed by hematocytometry (means + SEM) (B), CD8 + T cells, T reg cells, and NK cells pre- and postdose (7 days after treatment) (C) as well as CD8 + T–to–T reg cell ratios pre- and postdose (D) and over time (means + SEM) (E) as determined by flow cytometry in female cynomolgus monkeys ( n = 3) treated with Alb–IL-2 RNA-LNP (Alb–IL-2) or Alb–IL-2var RNA-LNP (Alb–IL-2var) (60 or 180 μg/kg). Empty LNPs (lipid dose adapted to 120 μg of RNA/kg) served as control. LLOQ, lower limit of quantification.

Article Snippet: Genes encoding the different IL-2 variants with an N-terminal His 6 -tag were synthesized and cloned into pTwist_CMV_BetaGlobin_WPRE_Neo expression vector (Twist Bioscience).

Techniques: Injection, Flow Cytometry, Control

Journal: Frontiers in Physiology

Article Title: Interleukin-13 R130Q aggravates airway smooth-muscle phenotypic switching and promotes airway remodeling in severe asthma

doi: 10.3389/fphys.2026.1906184

Figure Lengend Snippet: Effects of IL-13 130R and IL-13 130Q on pulmonary inflammatory responses. (A) Total cell counts in bronchoalveolar lavage fluid (BALF; n = 8 per group); differential leukocyte counting was not performed. (B–D) Relative mRNA expression levels of IL-4 (B) , IL-5 (C) , and IL-13 (D) in lung tissue (n = 8 per group). Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01. IL, interleukin; OVA, ovalbumin; SEM, standard error of the mean.

Article Snippet: According to the instructions of the manufacturer for the IL-4 and IL-5 ELISA kits (RayBiotech, Peachtree Corners, GA, USA), 100 μL of each sample was added to pre-coated wells and incubated at room temperature for 2.5 h. The wells were then washed three times for 5 min each, and blank wells served as negative controls.

Techniques: Expressing

Journal: Frontiers in Physiology

Article Title: Interleukin-13 R130Q aggravates airway smooth-muscle phenotypic switching and promotes airway remodeling in severe asthma

doi: 10.3389/fphys.2026.1906184

Figure Lengend Snippet: Effects of IL-13 130R and IL-13 130Q on human bronchial smooth muscle cell (hBSMC) phenotypes (n = 4 per group). (A) CCK-8-derived proliferative/metabolic activity. (B) Representative Fluo-3/AM flow-cytometry histograms and median fluorescence intensity quantification of the acetylcholine-induced intracellular calcium response. (C, D) IL-5 (C) and IL-4 (D) concentrations in culture supernatants. Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01. IL, interleukin; SEM, standard error of the mean.

Article Snippet: According to the instructions of the manufacturer for the IL-4 and IL-5 ELISA kits (RayBiotech, Peachtree Corners, GA, USA), 100 μL of each sample was added to pre-coated wells and incubated at room temperature for 2.5 h. The wells were then washed three times for 5 min each, and blank wells served as negative controls.

Techniques: CCK-8 Assay, Derivative Assay, Activity Assay, Flow Cytometry, Fluorescence

Journal: Frontiers in Physiology

Article Title: Interleukin-13 R130Q aggravates airway smooth-muscle phenotypic switching and promotes airway remodeling in severe asthma

doi: 10.3389/fphys.2026.1906184

Figure Lengend Snippet: Effects of IL-13 130R and IL-13 130Q on pulmonary inflammatory responses. (A) Total cell counts in bronchoalveolar lavage fluid (BALF; n = 8 per group); differential leukocyte counting was not performed. (B–D) Relative mRNA expression levels of IL-4 (B) , IL-5 (C) , and IL-13 (D) in lung tissue (n = 8 per group). Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01. IL, interleukin; OVA, ovalbumin; SEM, standard error of the mean.

Article Snippet: According to the instructions of the manufacturer for the IL-4 and IL-5 ELISA kits (RayBiotech, Peachtree Corners, GA, USA), 100 μL of each sample was added to pre-coated wells and incubated at room temperature for 2.5 h. The wells were then washed three times for 5 min each, and blank wells served as negative controls.

Techniques: Expressing

Journal: Frontiers in Physiology

Article Title: Interleukin-13 R130Q aggravates airway smooth-muscle phenotypic switching and promotes airway remodeling in severe asthma

doi: 10.3389/fphys.2026.1906184

Figure Lengend Snippet: Effects of IL-13 130R and IL-13 130Q on human bronchial smooth muscle cell (hBSMC) phenotypes (n = 4 per group). (A) CCK-8-derived proliferative/metabolic activity. (B) Representative Fluo-3/AM flow-cytometry histograms and median fluorescence intensity quantification of the acetylcholine-induced intracellular calcium response. (C, D) IL-5 (C) and IL-4 (D) concentrations in culture supernatants. Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01. IL, interleukin; SEM, standard error of the mean.

Article Snippet: According to the instructions of the manufacturer for the IL-4 and IL-5 ELISA kits (RayBiotech, Peachtree Corners, GA, USA), 100 μL of each sample was added to pre-coated wells and incubated at room temperature for 2.5 h. The wells were then washed three times for 5 min each, and blank wells served as negative controls.

Techniques: CCK-8 Assay, Derivative Assay, Activity Assay, Flow Cytometry, Fluorescence