csf 1  (Miltenyi Biotec)

Journal: bioRxiv

Article Title: Noncanonical CD11b on CD8 + T cells promotes cytotoxic effector function

doi: 10.64898/2026.10.07.757268

Figure Lengend Snippet: a, Representative flow cytometry plot of CD11b expression on OT-I CD8 + T cells 10 days after in vitro stimulation. Representative of four independent experiments. b, Experimental schematic (left) and real-time cytotoxicity assay (xCELLigence; right) evaluating B16-OVA tumor cell lysis when cultured alone or treated OT-I CD11b⁻ or OT-I CD11b⁺ CD8 + T cells (E:T = 0.25:1). Representative of three independent experiments; replicate-level data are shown in . c, Experimental workflow for generating CD11b-deficient NY-ESO-1-specific human CD8⁺ T cells. Cells were transduced with an NY-ESO-1-specific TCR, selected for CD11b expression, edited using CRISPR–Cas9, expanded and assessed for antitumor activity in real-time cytotoxicity assays. d, Representative flow cytometry plots showing CD11b expression in control-electroporated (CD11b + ctrl) and ITGAM-targeted (CD11b KO) NY-ESO-1-specific CD8 + T cells (left), and real-time cytotoxicity assay (xCELLigence; right) evaluating SK-MEL28-NY-ESO-1 tumor cell lysis when cultured alone, or with CD11b + ctrl or CD11b KO NY-ESO-1 CD8 + T cells (E:T = 0.25:1). Representative of three independent experiments; CD11b knockout efficiency and two additional independent cytotoxicity experiments are shown in and , respectively. e, Representative flow cytometry plot (left) and quantification (right) of CD11b expression in untransduced (UTD) or 19BBz cells after 10 days of in vitro expansion (n = 7 donors). f, Real-time cytotoxicity assay (xCELLigence) evaluating SK-MEL28-CD19t tumor cell lysis when cultured alone or with CD11b + or CD11b⁻ 19BBz CAR-T cells (E:T = 0.25:1). Representative of three independent experiments; replicate-level data are shown in . P values were calculated using two-tailed, paired Student’s t test ( e ).

Article Snippet: Unless otherwise stated, T cells were activated with anti-CD3/CD28 Dynabeads (Thermo Fisher Scientific, #11132D) at a bead-to-cell ratio of 1:1 in CST OpTimizer T Cell Expansion medium (Thermo Fisher Scientific, #A1048501) containing 5% CTS Immune Cell Serum Replacement (Thermo Fisher Scientific, #A2596101) and recombinant human IL-2 (100 IU/mL; Miltenyi Biotec, #130-097-748).

Techniques: Flow Cytometry, Expressing, In Vitro, Cytotoxicity Assay, Lysis, Cell Culture, Transduction, CRISPR, Activity Assay, Control, Knock-Out, Two Tailed Test, FACS