Journal: Alzheimer's & Dementia

Article Title: Fibrillar tau alters cerebral endothelial cell metabolism, vascular inflammatory activation, and barrier function in vitro and in vivo

doi: 10.1002/alz.70077

Figure Lengend Snippet: Metabolic alterations induced by protofibrillar tau in association with pro‐inflammatory endothelial cell activation. A, Representative trace for glycolysis stress test ECAR measurement using a Seahorse XF96 extracellular flux analyzer in hCMECs treated with aggregated tau for 24 hours. B, ECAR analysis indicates increased glycolysis (top) and glycolytic capacity (bottom) in the presence of 5 or 25 nM tau for 24 hours. C, Representative ECAR trace for glycolysis stress test in hCMECs treated with aggregated tau for 3 hours. D, ECAR indicates increased glycolysis (top) and glycolytic capacity (bottom) after 3 hours of protofibrillar tau challenge. E, Expression of adhesion markers VCAM‐1 and ICAM‐1 at 24 hours post treatment with 5 and 25 nM tau, as determined by western blot analysis. F, Expression of adhesion markers VCAM‐1 and ICAM‐1 at 3 hours post treatment with 5 and 25 nM tau. G, Increased IL‐4, IL‐10, TNF‐α, and IFN‐γ release after 25 nM tau treatment for 3 hours, determined by MSD proinflammatory panel analysis. H, Increased levels of IL‐4 and increasing trends of IL‐6 released in the media of hCMECs treated with 25 nM protofibrillar tau for 24 hours. I, Treatment with α‐IL‐4 does not decrease VCAM‐1 expression in hCMECs at 24 hours post‐tau treatment. J, TEER measurements during co‐treatment with tau protofibrils (25 nM) and a neutralizing antibody against IL‐4 (α‐IL‐4) only demonstrate a partial recovery against tau‐mediated loss of barrier resistance. Statistical significance established by one‐way analysis of variance; (WB), Seahorse (SH), and Tukey post hoc test, multiple unpaired t test (MSD). Significant P values are reported in the graphs. ECAR, extracellular acidification rate; hCMEC, human cerebral microvascular endothelial cell; ICAM‐1, intercellular adhesion molecule 1; IFN‐γ, interferon gamma; IL, interleukin; MSD, Meso Scale Discovery; TEER, trans‐endothelial electrical resistance; TNF‐α, tumor necrosis factor alpha; VCAM‐1, vascular cell adhesion molecule 1; WB, western blot.

Article Snippet: All experimental treatments were prepared in EBM‐2 containing 1% FBS, including: pre‐aggregated 1N4R tau fibrils, tau monomers, the arachidonic acid–containing vehicle, human IL‐4 antibody (R&D Systems, MAB304; αIL‐4, 1:100), heptelidic acid (abcam, ab144269, 100 nM), 2‐deoxyglucose (Sigma D8375, 1 mM), and human tau fibrils.

Techniques: Activation Assay, Expressing, Western Blot

Journal: Alzheimer's & Dementia

Article Title: Fibrillar tau alters cerebral endothelial cell metabolism, vascular inflammatory activation, and barrier function in vitro and in vivo

doi: 10.1002/alz.70077

Figure Lengend Snippet: Modulation of glycolytic metabolism reverts fibrillar tau‐mediated loss of barrier resistance. A, HA decreases tau‐mediated IL‐4 production in hCMECs 24 hours after treatment. B, Western blot analysis of hCMECs co‐treated with tau and HA reveals a decrease in VCAM‐1 expression in the presence of the glycolysis inhibitor. C, TEER measurement after co‐treatment with protofibrillar tau and 100  nM HA indicates that blocking glycolysis prevents tau‐mediated loss of barrier resistance. D, 2‐deoxyglucose partially rescues tau‐mediated loss of barrier resistance after 3 hours of tau treatment. Statistical significance established by one‐way ANOVA (WB), and Tukey post hoc test, two‐way ANOVA (Electrical Cell Impedance Sensing‐Zθ). Significant P values are reported in the graphs. ANOVA, analysis of variance; hCMEC, human cerebral microvascular endothelial cell; HA, heptelidic acid; IL, interleukin; TEER, trans‐endothelial electrical resistance; VCAM‐1, vascular cell adhesion molecule 1; WB, western blot

Article Snippet: All experimental treatments were prepared in EBM‐2 containing 1% FBS, including: pre‐aggregated 1N4R tau fibrils, tau monomers, the arachidonic acid–containing vehicle, human IL‐4 antibody (R&D Systems, MAB304; αIL‐4, 1:100), heptelidic acid (abcam, ab144269, 100 nM), 2‐deoxyglucose (Sigma D8375, 1 mM), and human tau fibrils.

Techniques: Western Blot, Expressing, Blocking Assay

Journal: Alzheimer's & Dementia

Article Title: Fibrillar tau alters cerebral endothelial cell metabolism, vascular inflammatory activation, and barrier function in vitro and in vivo

doi: 10.1002/alz.70077

Figure Lengend Snippet: PS19 mice demonstrate vascular metabolic dysfunction and EC activation at 3 months. A, ATP production rate assay reveals increased glycolytic ATP production (left) and decreased mitochondrial ATP production rate (middle) in PS19 mice cerebral microvessels, compared to WT mice. Representative ATP production rates by percentage (right). B, Representative trace for Mito Stress Test OCR of extracted vessels reveals deficits in mitochondrial respiratory function in PS19 mice. C, OCR measurement in extracted microvessels demonstrates decreased basal respiration rates in PS19 mice. D–G, Western blot analysis reveals increased VCAM‐1 (H) and GLUT1 (I) expression, as well as increases in total (pro‐MMP2 + cleaved‐MMP2; left) and cleaved (right) MMP2 (J) in PS19 mice cortices. H, Increased pro‐inflammatory cytokine production (IL‐6, IL‐5, IL‐1β, and anti‐inflammatory cytokine IL‐10), by extracted cerebral microvessels. Statistical significance established by Student t test (WB, SH, MSD). Significant p values or trends are reported in the graphs. ATP, adenosine triphosphate; EC, endothelial cell; IL, interleukin; GLUT1, glucose transporter 1; MMP2, matrix metalloproteinase 2; MSD, Meso Scale Discovery; OCR, oxygen consumption rate; SH, Seahorse; VCAM‐1, vascular cell adhesion molecule 1; WB, western blot WT, wild type.

Article Snippet: All experimental treatments were prepared in EBM‐2 containing 1% FBS, including: pre‐aggregated 1N4R tau fibrils, tau monomers, the arachidonic acid–containing vehicle, human IL‐4 antibody (R&D Systems, MAB304; αIL‐4, 1:100), heptelidic acid (abcam, ab144269, 100 nM), 2‐deoxyglucose (Sigma D8375, 1 mM), and human tau fibrils.

Techniques: Activation Assay, Western Blot, Expressing

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