Journal: Transplantation Direct

Article Title: Kinetics of Alloantigen-Specific Regulatory CD4 T Cell Development and Tissue Distribution After Donor-Specific Transfusion and Costimulatory Blockade

doi: 10.1097/TXD.0000000000000580

Figure Lengend Snippet: Abrogation of linked suppression by co-injection of blocking antibodies. Self-B6 regulation responses were variable, as measured by linked suppression of recall tv-DTH response on day 14. No significant differences in recovery of TT/DT response from linked suppression by self (B6) antigens were observed on day 14 with IL10, TGFβ or IL35 neutralization, although a trend ( P = 0.06, P = 0.09, and P = 0.06, respectively) was observed. However, allo (CBA)-specific linked suppression responses on day 35 were clearly reversed by the IL10, TGFβ or IL35 cytokine-neutralizing antibodies (all P < 0.001). The co-injection of αIL4 antibody did not elicit a significant LS reversal effect relative to IgG isotype control. Data were presented as mean with SEM from 3 to 6 mice in each group (Open symbols, wild type B6 mice; closed symbols, Foxp3/YFP, TdTomRed/Ebi3 reporter B6 mice), and analyzed by Student t test (*** P < 0.001).

Article Snippet: Coinjected blocking antibodies were: αTGFβ (BD Pharmigen) (10 μg), αIL10 (R&D Systems) (10 μg), a combination of αp35 (R&D Systems) + αEbi3 (provided by Dr. Vignali, University of Pittsburgh) (1 μg of each), and αIL4 (BD Bioscience) (10 μg).

Techniques: Injection, Blocking Assay, Neutralization

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 Promotes Type-2 Immune Signature in CT26 Tumor Microenvironment. (A) Frequency of CT26 tumor-infiltrating IL4-eGFP + CD4 + T cells (TH2) in VML-injured (untreated and ECM-treated) IL4-eGFP reporter (4Get) mice. CD4 + CD25 + (Tregs) were removed before TH2 gating (flow plot % are out of parent CD4 + CD25 - , which differs from bar graph depicting % out of all CD4 + ). (B) Differential gene expression and (C) normalized RNA transcript counts per million for type-2 cytokine genes from bulk RNAseq of CT26 tumor-infiltrating CD3 + T cells from uninjured and VML injured (untreated and ECM-treated) mice. (D) Intra-tumoral density of type-2 cytokine (IL4, IL5, IL13)-producing CT26 tumor-infiltrating CD4 + T cells from uninjured and VML injured (untreated and ECM-treated) mice following ex vivo cell stimulation. (E) Frequency of CT26 tumor-infiltrating IL4-eGFP + CD11b + myeloid cells in VML injured (untreated and ECM-treated) 4Get mice. (F) Differential gene expression from bulk RNAseq of CT26 tumor-associated F480 + macrophages from untreated and ECM-treated VML injured mice. (G) Polarization of CT26 tumor-associated macrophages towards regenerative R1 (CD9 + CD301b + MHCII + ) phenotype in uninjured and VML injured (untreated and ECM-treated) mice. (Statistics) 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 ( A, E ). For >2 groups, data was analyzed using an ordinary one-way ANOVA with Tukey’s multiple comparisons test ( D, G ). Bulk RNAseq: Negative-binomial distribution using DeSeq2 with FDR correction for multiple testing (adjusted p-value thresholds indicated on plots) ( B, F ). No statistical analysis was performed on normalized transcript counts ( C ). NS: Not significant p>0.05, * p<0.05, ** p<0.01.

Article Snippet: IL4 was neutralized using αIL4 antibody (clone 11B11, BioXCell BE0045) delivered via IP injection prepared in sterile dilution buffer ( InVivo Pure pH 7.0, BioXCell IP0070).

Techniques: Gene Expression, RNA sequencing, Ex Vivo, Cell Stimulation, 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: 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: IL4 was neutralized using αIL4 antibody (clone 11B11, BioXCell BE0045) delivered via IP injection prepared in sterile dilution buffer ( InVivo Pure pH 7.0, BioXCell IP0070).

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