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: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Concentration-dependent change in the humoral inflammatory response following incubation with Escherichia coli ( E. coli ) in the ex vivo whole blood model. a Absolute plasma concentrations of IL-6, IL-8, and MMP9 determined by enzyme-linked immunosorbent assay. b Normalized values and EC 50 curve fit by BuC=0% and 50 000 CFU/ml E. coli= 100%, respectively, for IL-6, IL-8, and MMP9 as indicated by EC 50 (%) on the respective Y-axis. BuC indicates buffer control after 60 min incubation; numbers on the X-axis indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation; LPS indicates lipopolysaccharide (LPS) 100 ng/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, comparing all shown concentrations of E. coli bacteria and 100 ng/ml LPS with BuC. P -values are indicated above the respective data points. ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Concentration Assay, Incubation, Ex Vivo, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, Bacteria

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Diagnostic performance for the detection of bacteremia, analyzing the neutrophil phenotype by determining the median fluorescence intensity (MFI) and the cellular response capacity (CRC) in comparison with traditional markers of humoral inflammation (IL-6, IL-8, MMP9). a Comparison of receiver operating characteristic (ROC) at 10,000 CFU/ml Escherichia coli ( E. coli ) with the respective 95% confidence interval (CI) and P -value, and half-maximal effective concentration (EC 50 ) as a function of the E. coli concentration. b Detailed comparison of the EC 50 as a function of the E. coli concentration. c Exemplary comparison of EC 50 curve fit after normalization as indicated by EC 50 (%) on the respective Y-axis to BuC=100% and 50 000 CFU/ml E. coli =0% for the humoral marker IL-6 (the IL-6 values were multiplied by −1 before EC 50 calculation to facilitate comparability with the CRC) and the change in neutrophil phenotype represented by CD11b CRC. BuC indicates buffer control after 60 min incubation; numbers on the X-axis of c indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, evaluating the EC 50 of IL-8, MMP9, the MFI, and CRC of CD10, CD11b, and CD62L in comparison to the EC 50 of IL-6. P -values are indicated above the respective data points. ⁎ P <0.05. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Diagnostic Assay, Fluorescence, Comparison, Concentration Assay, Marker, Control, Incubation, Bacteria

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Clinical specifications and parameters over all time points of the sepsis cohort. a Suspected infection cause of sepsis. b Distribution of the individual score points of the Sequential Organ Failure Assessment (SOFA) score. c Total SOFA score. d-h Traditional and humoral markers of inflammation: leukocytes and neutrophil-lymphocyte ratio ( d ), C-reactive protein (CRP) and procalcitonin (PCT) ( e ), interleukin-6 (IL-6) and interleukin-8 (IL-8) ( f ), serum amyloid A (SAA) and calprotectin ( g ), matrix metallopeptidase 9 (MMP9) and myeloperoxidase (MPO) ( h ). Values are shown as median and interquartile range. n =14. CNS. Central nervous system; HV. Healthy volunteers.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Infection

Journal: Bioactive Materials

Article Title: Dynamic feedback BacGuard anchors microbial metabolism to host symbiosis in real-time ulcerative colitis therapy

doi: 10.1016/j.bioactmat.2026.05.060

Figure Lengend Snippet: BacGuard promotes colon tissue repairment. a) Scheme of microbiota-dependent epithelial repair mechanism orchestrated by BacGuard. b) Short-chain fatty acid (SCFA) profile alterations. n = 6. c) BacGuard-induced probiotic proliferation and d) quantitative results. n = 3. e) Immunofluorescence analysis of ILC3 (ROR γt + CD3 − cells) in colon tissue. f) Flow cytometric analysis of lamina propria lymphocytes (ROR γt + ). n = 3. g) Concentration of IL-22 in MNK-3 cells. n = 3. h) Representative PAS-staining (upper panel) and MUC-2 immunohistochemistry (lower panel) images of colon tissues. n = 5. ns, not significant; ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001.

Article Snippet: For the detection of IL-22 and LPS concentrations, the Mouse IL-22 Precoated ELISA Kit (DAKEWE, China) and Mouse LPS ELISA Kit (JONLNBIO, China) were employed correspondingly.

Techniques: Immunofluorescence, Concentration Assay, Staining, Immunohistochemistry