Journal: bioRxiv

Article Title: Molecular Imaging of the TGF-β Activating Integrin αvβ6 Detects Chronic Lung Allograft Dysfunction

doi: 10.64898/2026.01.07.698265

Figure Lengend Snippet: (A) Flow cytometric analysis of POD 16-tolerant and CLAD allografts with (A) representative contour plots gated on live CD45-allograft cells (N=5/group) and (B) Intracellular Sftpc expression analysis on CLAD allograft CD326+ αvβ6+ cells. The histogram shown is representative of three independent experiments. (C) Plot of POD 16 allograft numbers of live CD45-CD326+ αvβ6+ cells shown with a mean ± standard deviation from the mean for N=5 per group. Representative immunohistochemical stains with αvβ6 specific antibodies with indicated magnification and scales (N≥4/group).

Article Snippet: Sections were then stained with 1:200 αvβ6 antibodies (bs-5791R, Bioss for mouse tissue; AB00891-23.0, Absolute Antibody for human tissue) overnight at 4°C.

Techniques: Expressing, Standard Deviation, Immunohistochemical staining

Journal: bioRxiv

Article Title: Molecular Imaging of the TGF-β Activating Integrin αvβ6 Detects Chronic Lung Allograft Dysfunction

doi: 10.64898/2026.01.07.698265

Figure Lengend Snippet: αvβ6 immunohistological staining of allografts from recipients that underwent club cell injury and received (A) CCL2 neutralizing Abs, (B) CD8α depleting Abs, respectively, and (A, B) Control Ig injections followed by POD 16 [ 64 Cu]Cu-DOTA-A20-K16R PET/CT imaging. Images shown are representative of N≥4 per group. Scale bars: 400 μm for 10x, 100 μm for 40x.

Article Snippet: Sections were then stained with 1:200 αvβ6 antibodies (bs-5791R, Bioss for mouse tissue; AB00891-23.0, Absolute Antibody for human tissue) overnight at 4°C.

Techniques: Staining, Control, Positron Emission Tomography-Computed Tomography, Imaging

Journal: bioRxiv

Article Title: Molecular Imaging of the TGF-β Activating Integrin αvβ6 Detects Chronic Lung Allograft Dysfunction

doi: 10.64898/2026.01.07.698265

Figure Lengend Snippet: (A, Left Panel) Representative autoradiographic images following incubation of [ 64 Cu]Cu-DOTA-A20-K16R with human explanted lung tissue sections from a patient with CLAD and Control lung tissue from a donor lung obtained prior to transplantation. Preincubation with an excess of a cold probe sharply reduces autoradiographic activity (Cold Probe/CLAD). (A, Right Panel) corresponding trichrome-stained lung sections that were incubated with radiotracer. Images are representative of independently conducted experiments, with (B) violin plots showing combined data from 3 explanted CLAD transplants with or without cold-probe blockade and 3 control lungs per group. Two-sided Mann-Whitney U t-test *p<0.05, ***p<0.001. Representative αvβ6 (C) immunohistochemical and (D) immunofluorescent staining of human explanted lungs with CLAD and control lungs (N=3/group). Arrows denote clusters of cells expressing αvβ6. Scale bars: 400 μm for 10x, 100 μm for 40x.

Article Snippet: Sections were then stained with 1:200 αvβ6 antibodies (bs-5791R, Bioss for mouse tissue; AB00891-23.0, Absolute Antibody for human tissue) overnight at 4°C.

Techniques: Incubation, Control, Transplantation Assay, Activity Assay, Staining, MANN-WHITNEY, Immunohistochemical staining, Expressing

Journal: bioRxiv

Article Title: UC-associated autoantibodies to αvβ6 inhibit mucosal TGFβ activation and predispose to intestinal inflammation

doi: 10.64898/2026.03.12.709585

Figure Lengend Snippet: ( A-B ) Serum anti-αvβ6 IgG ( A ) and IgA ( B ) quantified by ELISA in healthy subjects (HS; n = 338) and ulcerative colitis (UC; n = 194). Dashed lines indicate OD thresholds for autoantibody positivity, determined by receiver operating characteristic (ROC) curve analysis using the highest Youden’s index. ( C-D ) Concentrations of anti-αvβ6 IgG (HS, n=10; UC n=153) ( C ) and IgA (HS, n=4; UC n=61) ( D ) in sera were interpolated from monoclonal antibody standard curves; samples below the lower limit of quantification (LLOQ) are indicated and were excluded from concentration-based statistical analysis. ( E-F ) Correlation between serum anti-αvβ6 IgG (left) or IgA (right) OD values and Simple Clinical Colitis Activity Index (SCCAI) scores (n=171) ( E ) or serum C-reactive protein (CRP) levels (n=132) ( F ) in UC donors. ( G ) Representative IgG ELISPOT images (left) and quantification (right) of αvβ6-specific IgG-producing cells in peripheral blood mononuclear cells. ( H ) Representative IgG ELISPOT images (left) and quantification (right) of αvβ6-specific IgG and IgA-producing cells in colonic biopsies. Wells coated without αvβ6 served as negative controls. ( I, J ) Anti-αvβ6 IgG ( I ) and IgA ( J ) levels measured in supernatants from overnight-cultured colonic biopsies; dotted lines indicate lower limits of detection (LLOD). Data are shown as individual data points with medians ( A-D, G-J ) or with best-fit lines for visualization purposes only ( E-F ). Two-group comparisons were performed using two-tailed Mann-Whitney tests ( A-D, G, I, J ). Correlations were assessed using two-tailed Spearman rank correlation ( E-F ).

Article Snippet: For induction of αvβ6 autoantibodies, C57BL/6J mice (Jackson Laboratory) were immunized subcutaneously with recombinant human αvβ6 protein (WuXi Biologics) emulsified in Complete Freund’s Adjuvant (CFA; InvivoGen), followed by booster immunization in Incomplete Freund’s Adjuvant (IFA; InvivoGen) two weeks later.

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Activity Assay, Enzyme-linked Immunospot, Cell Culture, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: UC-associated autoantibodies to αvβ6 inhibit mucosal TGFβ activation and predispose to intestinal inflammation

doi: 10.64898/2026.03.12.709585

Figure Lengend Snippet: (A) Adhesion of HT-29 cells to latent TGFβ (L-TGFβ) in the presence of anti-αvβ6 blocking antibody 3G9, HS serum, or UC serum, normalized to untreated controls. (B) Inhibition of cell adhesion to L-TGFβ by individual donor UC serum (left axis) plotted against corresponding serum anti-αvβ6 IgG levels (right axis). (C) Representative flow cytometry histograms showing surface expression of αvβ6 on T84 cells stained with 3G9 (green) or secondary-only control (gray). (D) Schematic of the in vitro assay used to measure αvβ6-dependent activation of L-TGFβ by T84 cells and signaling to HEK-Blue TM TGFβ reporter cells. (E) αvβ6-dependent activation of latent TGFβ by T84 cells measured by HEK-Blue reporter activity (OD620); dotted line indicates mean reporter activity across HS controls. Data represent mean ± SD and statistical significance determined using unpaired two-tailed t test with Welch’s correction (A) , individual data points (B) or mean ± SEM (E)

Article Snippet: For induction of αvβ6 autoantibodies, C57BL/6J mice (Jackson Laboratory) were immunized subcutaneously with recombinant human αvβ6 protein (WuXi Biologics) emulsified in Complete Freund’s Adjuvant (CFA; InvivoGen), followed by booster immunization in Incomplete Freund’s Adjuvant (IFA; InvivoGen) two weeks later.

Techniques: Blocking Assay, Inhibition, Flow Cytometry, Expressing, Staining, Control, In Vitro, Activation Assay, Activity Assay, Two Tailed Test

Journal: bioRxiv

Article Title: UC-associated autoantibodies to αvβ6 inhibit mucosal TGFβ activation and predispose to intestinal inflammation

doi: 10.64898/2026.03.12.709585

Figure Lengend Snippet: (A) Serum anti-human (left) and anti-mouse (right) αvβ6 IgG in αvβ6-immunized and adjuvant-only (n=8 per group) mice quantified by ELISA following primary immunization and boost. (B) Inhibition of αvβ6-dependent adhesion to L-TGFβ by serial dilutions of serum from αvβ6-immunized (n=6) or adjuvant-only (n=4) mice. (C) Body weight following primary immunization and boost. (D) Representative colonic H&E at baseline. (E, F) Representative flow cytometry plots and frequencies of CD8αß + T cells among TCRß + T cells (E) and CD103 + CD69 + T cells among CD8ß + T cells (F) from IELs. (H-K) Following DSS-induced colitis, body weight (H) , survival (I) , colon length (J) , and representative H&E-stained colon sections at day 14 post-DSS (K) . Body weight data (mean ± SEM) were analyzed using a mixed-effects model (REML; p = 0.0224). Survival was analyzed using Kaplan–Meier curves and compared by the log-rank (Mantel–Cox) test (p=0.22). Flow cytometry data are shown as mean ± SD and are representative of a single experiment (of two independent experiments) conducted with n = 4 adjuvant-only and n = 5 αvβ6-immunized mice per group. Statistical significance was determined using multiple unpaired t tests with Welch’s correction (F, H, J) or log-rank (Mantel–Cox) test (I) . *P < 0.05; **P < 0.01; ***P < 0.001

Article Snippet: For induction of αvβ6 autoantibodies, C57BL/6J mice (Jackson Laboratory) were immunized subcutaneously with recombinant human αvβ6 protein (WuXi Biologics) emulsified in Complete Freund’s Adjuvant (CFA; InvivoGen), followed by booster immunization in Incomplete Freund’s Adjuvant (IFA; InvivoGen) two weeks later.

Techniques: Adjuvant, Enzyme-linked Immunosorbent Assay, Inhibition, Flow Cytometry, Staining