Journal: bioRxiv

Article Title: PTPN1/2 inhibits alveolar macrophage-mediated control of lung metastasis

doi: 10.64898/2026.02.25.707995

Figure Lengend Snippet: A. C57BL/6 mice were subcutaneously implanted with 1 × 10⁶ CMT167 cells and treated daily with 20 mg/kg AC484 or vehicle. B. BALB/c mice were implanted with 1 × 10⁵ 4T1 cells in the mammary fat pad and treated as indicated. C. Representative H&E-stained and analysis markup images of lungs collected at the endpoint from A. Yellow, lung parenchyma; red, metastatic tumors; light blue, glass background. Scale bar = 2 mm. D. Tumor area quantification from C. Shown are mean ± SEM from 3 independent sections. E. Visible lung surface metastasis counts in CMT167-bearing mice at the endpoint. F. Representative H&E-stained and analysis markup images of lungs collected at the endpoint from B. Yellow, lung parenchyma; red, metastatic tumors; light blue, glass background. Scale bar = 2 mm. Arrows point to metastatic tumor regions. G. Tumor area quantification from F. Mean ± SEM from 3 sections. H. Surface metastases count in 4T1-bearing mice at the endpoint. I. Western blot of PTPN1/2 double knockout (dKO) and scrambled (Scr) control 4T1 cells treated overnight with IFNγ or IL-6 (100 ng/mL), in the presence or absence of 1 μM AC484. J. BALB/c mice implanted with dKO or Scr 4T1 cells and treated as indicated. Lung surface metastases quantified after India ink staining. K-M. NSG mice implanted with 4T1 or CMT167 cells and treated as indicated. Lung metastases were visualized and quantified separately for 4T1 ( L ) and CMT167 ( M ). N. C57BL/6 mice were treated with depleting antibodies (anti-CD8β, anti-NK1.1, or isotype control) starting on Day -1, followed by CMT167 cell implantation and AC484 treatment. Surface metastases were quantified at the endpoint. For all bar graphs, shown are individual animals (dots), with mean ± SEM (bars). Unpaired t-test for two-group comparisons; one-way ANOVA for multiple groups: ns, p > 0.05; * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001.

Article Snippet: For cell depletion, mice were administered i.p . injections of 200 μg anti-CD8β (clone 53.5.8, BioXcell) every four days to deplete CD8+ T cells, 200 μg anti-NK1.1 (clone PK136, BioXcell) every four days to deplete NK cells, 500 μg anti-CSF1R (clone AFS98, BioXcell) every other day to deplete interstitial macrophages ( , ), and 200 μg anti-IFNγ (clone XMG1.2, BioXcell) every three days to neutralize IFN-γ.

Techniques: Staining, Western Blot, Double Knockout, Control

Journal: bioRxiv

Article Title: Tissue Injury and Biomaterial Treatment Modulate Tumor Growth and Response to Immunotherapy

doi: 10.64898/2026.02.02.703323

Figure Lengend Snippet: VML Injury Impacts Anti-Tumor CD8 + T Cell Response. (A) Flow cytometric profiling of CT26 tumor-infiltrating CD8 + T cells, (B) PD1 expression on CD8 + T cells, and (C) CT26 tumor-specific CD8 + T cells marked by AH1-loaded MHC class I tetramer in uninjured and VML injured mice. (D) Single-cell RNA-sequencing (scRNAseq) of CT26 tumor-infiltrating T cells from uninjured and VML injured mice. UMAP depicting main T cell clusters (e.g., CD8, CD4, Treg, unconventional). (E) T cell UMAP separated by tumor-reactive AH1-Tetramer + CD8 + T cells (top) and other T cells (bottom) colored by cells from uninjured (gray) and VML injured (green) mice. (F) Differential gene expression and (G, H) GSEA using Hallmark or (I) curated list of T cell-related pathways with top leading edge genes comparing tumor-reactive AH1-Tetramer + CD8 + T cells of uninjured and VML injured mice. (J) Intracellular cytokine staining following ex vivo stimulation of IFNγ-producing effector cell types in CT26 tumors of uninjured and VML injured mice. (Statistics) Bar graphs: mean±SD. Normally distributed data (Shapiro-Wilk test, α=0.05) was analyzed using an unpaired two-tailed student t-test ( A-C, J ). Results representative of at least 2 independent experiments ( A-C ). NS: Not significant p>0.05, * p<0.05, ** p<0.01. UMAP: Uniform manifold approximation and projection. GSEA: Gene set enrichment analysis.

Article Snippet: CD8 + T cells were depleted using αCD8β antibody (clone 53-5.8, BioXCell BE0223) delivered via IP injections at 5mg/kg body weight prepared in sterile dilution buffer ( InVivo Pure pH 7.0, BioXCell IP0070).

Techniques: Expressing, Single Cell, RNA Sequencing, Gene Expression, Staining, Ex Vivo, Two Tailed Test

Journal: bioRxiv

Article Title: Tissue Injury and Biomaterial Treatment Modulate Tumor Growth and Response to Immunotherapy

doi: 10.64898/2026.02.02.703323

Figure Lengend Snippet: VML Injury Impacts Distant Tumor Growth by Engaging a Shared Tumor-Draining Lymph Node (tdLN). (A) Flow cytometric profiling of CD8 + T cells in CT26 tdLN, including effector memory differentiation (CD44 hi CD62L - ) and (B, C) activation (CD25 + , NKG2D + ) in uninjured and VML-injured mice. (D) The bilateral VML injury (square) was modified to a unilateral procedure performed on either the ipsilateral (triangle) or contralateral (inverted triangle) quadriceps muscle relative to the tumor-bearing flank. CT26 tumor growth kinetics of uninjured and VML-injured (bi-, ipsi-, and contra-lateral) mice (n=9-10). (E) Survival curve (volume >1500mm 3 or severe involuting ulceration) of CT26 tumor-bearing mice with ipsi-versus contra-lateral VML injury. (F) Inhibition of lymphocyte LN egress using FTY720 HCl (25 μg/mouse). Representative flow cytometry plots and quantification of CD3 + T cells in peripheral blood (collected at survival endpoint) to confirm successful sequestration of lymphocytes in LN/spleen. (G) CT26 tumor growth kinetics of uninjured and ipsilateral VML-injured mice treated with vehicle (solid) or FTY720 HCl (dashed) (n=9-10). (Statistics) Tumor growth curves: mean±SEM. Bar graphs: mean±SD ( D, E display earliest survival timepoint). For 2 groups, normally distributed data (Shapiro-Wilk test, α=0.05) was analyzed using an unpaired two-tailed student t-test ( B, C ); otherwise, a non-parametric two-tailed Mann-Whitney test was used ( A ). For >2 groups, data was analyzed using an ordinary one-way ( D ) or two-way ANOVA ( F, G ) with Tukey’s multiple comparisons test (only relevant comparisons shown). Survival: Kaplan-Meier curve with Log-Rank Mantel-Cox test ( E ). Results representative of at least 2 independent experiments ( D, G ). NS: Not significant p>0.05, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

Article Snippet: CD8 + T cells were depleted using αCD8β antibody (clone 53-5.8, BioXCell BE0223) delivered via IP injections at 5mg/kg body weight prepared in sterile dilution buffer ( InVivo Pure pH 7.0, BioXCell IP0070).

Techniques: Activation Assay, Modification, Inhibition, Flow Cytometry, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Tissue Injury and Biomaterial Treatment Modulate Tumor Growth and Response to Immunotherapy

doi: 10.64898/2026.02.02.703323

Figure Lengend Snippet: Biomaterial Injury Treatment-Induced Type-2 Immunity Also Develops in Tumor-Associated Tissues and Contributes to Delayed Tumor Growth. (A) Flow cytometric profiling of B and T lymphocytes, (b) Tregs (CD4 + FoxP3 + CD25 + ), and (C) CD8 + /Treg ratio in CT26 tumor-draining lymph nodes (tdLNs) of uninjured and VML-injured (untreated and ECM-treated) mice. (D) Frequency of IL4-eGFP + CD4 + T cells (TH2) in CT26 tdLNs of VML-injured (untreated and ECM-treated) 4Get mice. (E) Type-2 cytokine (IL4 and IL13) production by CD4 + T cells in CT26 tdLNs of uninjured and VML-injured (untreated and ECM-treated) mice following ex vivo cell stimulation. (F) Frequency of IL4-eGFP + SiglecF + eosinophils and CD4 + T cells in CT26 tumor-encapsulating adipose of VML-injured (untreated and ECM-treated) 4Get mice. (G) IL4 neutralization: CT26 tumor growth and survival curves of VML-injured (untreated and ECM-treated) mice treated with isotype (solid) or αIL4 antibody (1 mg/mouse initial dose followed by 0.5 mg/mouse maintenance) (dashed) (n=9-10). (Statistics) Tumor growth curves: mean±SEM. Bar graphs: mean±SD. For 2 groups, normally distributed data (Shapiro-Wilk test, α=0.05) was analyzed using an unpaired, two-tailed student t-test ( D, F ). For >2 groups, data was analyzed using an ordinary one-way ( A-C, E ) or two-way ( G - only D20 ) ANOVA with Tukey’s multiple comparisons test. Survival: Kaplan-Meier curve with Log-Rank Mantel-Cox test ( G ). NS: Not significant p>0.05, * p<0.05, ** p<0.01, *** p<0.001.

Article Snippet: CD8 + T cells were depleted using αCD8β antibody (clone 53-5.8, BioXCell BE0223) delivered via IP injections at 5mg/kg body weight prepared in sterile dilution buffer ( InVivo Pure pH 7.0, BioXCell IP0070).

Techniques: Ex Vivo, Cell Stimulation, Neutralization, Two Tailed Test

Journal: Molecular Therapy. Nucleic Acids

Article Title: mRNA vaccine expressing enterovirus D68 virus-like particles induces potent neutralizing antibodies and protects against infection

doi: 10.1016/j.omtn.2025.102731

Figure Lengend Snippet: mRNA vaccine expressing VLP induced cellular immunity, contributing to CD8 T + cell-dependent cross-protection against CVB3 (A) IFN-γ response in splenocytes. The mRNA vaccine (P1:3CD = 1:1 on weight, total 2 μg of mRNA) and IWV (2 μg of protein) immunizations were performed twice before the splenocytes were collected. The splenocytes were cultured in the absence (non-treat) or presence of a mixture of recombinant 3C and 3D, 3C, or 3D protein of EV-D68, and the IFN-γ in the culture supernatants were quantified by ELISA. Points are presented as individual data, and lines are presented as the median. Dotted lines indicate detection limits. (B) Experiment schema for (C–E). The P1 or 3CD mRNA vaccine (1 μg of mRNA per dose) immunizations were performed twice, and CVB3 challenge was started on day 30. For CD4 + or CD8 + T cell depletion, mice vaccinated with 3CD mRNA were administered anti-CD4 (αCD4), anti-CD8 (αCD8), or their isotype control antibodies 1 day before the challenge. (C) The weight change after CVB3 challenge. (D, E) Influence of CD4 + or CD8 + T cell depletion on the efficacy of EV-D68 3CD against CVB3 challenge. (C–E) Data are shown as means ± SD. Dotted line represents the humane endpoint. (A) ∗∗ p < 0.01; ns, not significant as indicated by Tukey’s test. (C) # p < 0.05; ## p < 0.01; ### p < 0.001; #### p < 0.0001 as indicated by Dunnett’s test compared with 3CD. Black symbols show significance of PBS against 3CD, and red symbols show significance of P1 against 3CD. (D, E) § p < 0.05; §§§ p < 0.001; as indicated by Šidák’s test compared with isotype control group.

Article Snippet: For CD8 + T cell depletion, 100 μg of anti-mouse CD8β antibody (catalog number #BE0223, clone: 53-5.8, Bio X Cell) or IgG1 isotype control (catalog number #BE0088, clone: HRPN, Bio X Cell) was administered intraperitoneally 1 day before the challenge.

Techniques: Expressing, Cell Culture, Recombinant, Enzyme-linked Immunosorbent Assay, Control