bite  (InvivoGen)

Journal: bioRxiv

Article Title: The T-cell niche tunes immune function through modulation of the cytoskeleton and TCR-antigen forces

doi: 10.1101/2024.01.31.578101

Figure Lengend Snippet: a , Schematic of the mechanism of action of blinatumomab. Blinatumomab is a bispecific T-cell engager (BiTE) that bridges CD3+ T-cells with CD19+ B-cells to induce cytolysis. b , Schematic of the blinatumomab DNA-based tension probe design (“blina-probe”). Blinatumomab is added to CD19-presenting tension probes to bridge CD3+ T-cells with CD19-presenting probes. c , Representative images and d , quantification of locked 4.7 pN tension signal of donor T-cells on blina-probes after a 24 hr treatment with UCM, SCM, or ACM. Circles (●) represent cells from donor 1, squares (▪) represent cells from donor 2, triangles (▴) represent cells from donor 3. Scale bar = 10 μm. Graphs represent means ± SD and data points represent fluorescence intensity measurements of individual cells. Statistics were calculated using a one-way ANOVA with a Tukey’s multiple comparisons test. **P < 0.0091, ****P < 0.0001, n = 3 independent donors.

Article Snippet: Blinatumomab (# bimab-hcd19cd3) and SIINFEKL-OVA peptide (# vac-sin) were purchased from InVivoGen.

Techniques: Fluorescence

Journal: bioRxiv

Article Title: The T-cell niche tunes immune function through modulation of the cytoskeleton and TCR-antigen forces

doi: 10.1101/2024.01.31.578101

Figure Lengend Snippet: a-b , Representative ImageStream flow cytometer images of blinatumomab-mediated immunological synapse formation after a 24 hr treatment with UCM, SCM or ACM. T-cells were stained with LAMP-1, LFA-2, and SirActin to show markers of immunological synapse formation. Nalm6 B-cells were stained with cell tracker violet. Scale bars = 7 μm. c , Fold-increase of immunological synapse formation in blinatumomab-treated samples compared to vehicle-treated samples. d , Blinatumomab-mediated cytolysis of Nalm6 B-cells after a 24 hr or e , 48 hr treatment with UCM, SCM, or ACM. %Lysis was calculated using 100 x (% blinatumomab-treated sample [Dead Nalm6] - %vehicle-treated sample [Dead Nalm6]). 24 hr: UCM v. SCM * P = 0.0151, UCM v. ACM * P = 0.0151, SCM v. ACM *** P = 0.0004. 48 hr: UCM v. SCM P = 0.1201, UCM v. ACM P = 0.0971, SCM v. ACM *** P = 0.0064. Graphs represent means ± SD. Data points represent individual donors. Statistics were calculated using a one-way ANOVA with a Tukey’s multiple comparisons test. n = 2-3 independent donors.

Article Snippet: Blinatumomab (# bimab-hcd19cd3) and SIINFEKL-OVA peptide (# vac-sin) were purchased from InVivoGen.

Techniques: Flow Cytometry, Staining, Lysis