anti mouse cd3  (Bio-Rad)

Journal: Cell reports

Article Title: De-coupling immune parameters and toxicity associated with IL-12 agonism

doi: 10.1016/j.celrep.2025.115840

Figure Lengend Snippet: (A–C) C57BL/6 (WT) and IFN-γ −/− mice ( n = 5 mice/group) received daily i.p. injections of IL-12 Fc, IL-12 3x Ala Fc, or an IgG2a Fc as a control and mice were euthanized on day 4 and lungs harvested for analysis of myeloid cells (CD3 − , Ly6G − , B220 − , Sirtpa + , CD11c − ) by flow cytometry. (A) Quantification of total monocyte and macrophage cells in the lung for 1 representative experiment of 3 performed. Data are presented as x ‾ ± S E M with statistical significance determined by two-way ANOVA and Tukey’s multiple correction (* p < 0.05 and ** p < 0.005). (B) Representative flow plot of macrophage and monocyte populations depicting (1) Ly6C low , (2) Ly6C high , and (3) MHCII + inflammatory monocytes, and (4) Ly6C − MHCII + macrophages. (C) Total numbers ( x ‾ ± S E M ) of Ly6C low , Ly6C high , and Ly6C + MHCII + inflammatory monocytes and Ly6C − MHCII + macrophages, with statistical significance determined by two-way ANOVA and Tukey’s multiple correction (* p < 0.05, ** p < 0.005, *** p < 0.001, and **** p < 0.0001). (D) Representative histograms of PD-L1, CD80, and CD86 on inflammatory monocytes (Ly6C + MHCII + ) with quantification of the geometric mean fluorescence intensity. Data are presented as x ‾ ± S E M from a single experiment with n = 5 per group, and similar results were seen in a repeat experiment. Statistical significance was determined by one-way ANOVA and Tukey’s multiple correction (*** p < 0.001 and **** p < 0.0001).

Article Snippet: Armenian hamster anti-mouse CD3 (clone: 145-2C11) , BioXcell , Catalog: BE0001-1; RRID: AB_1107634.

Techniques: Control, Flow Cytometry, Fluorescence

Journal: Cell reports

Article Title: De-coupling immune parameters and toxicity associated with IL-12 agonism

doi: 10.1016/j.celrep.2025.115840

Figure Lengend Snippet: IFN-γ Thy1.1 reporter mice received daily i.p. injections of IL-12 Fc, IL-12 3x Ala Fc, or an IgG2a Fc fragment as a control and were euthanized after 7 days, and the spleen and liver were analyzed to characterize the natural killer (NK) and innate-like cell 1 (ILC1) response. (A) Quantification of the total splenic NK and ILC1 cell populations ( n = 5 per group, x ‾ ± S E M ). NK cells are defined as live, singlets, CD3 − , NK1.1 + Tbet + Eomes + and ILC1s are defined as live, singlets, CD3 − , NK1.1 + Tbet + Eomes − . Data shown are representative of 3 experiments performed. Statistical significance was determined by one-way ANOVA and Tukey’s multiple correction; (*** p < 0.001 and **** p < 0.0001). (B and E) Representative flow plots depicting splenic and lung NK cells (defined as Eomes high and Eomes low populations) and splenic ILC1 cells after gating down to live, singlets, CD3 − , NK1.1 + . (C and F) Representative flow plot depicting bound IL-12 agonist x Thy1.1 expression for each subset of splenic NK cells (defined as Eomes high and Eomes low populations) and ILC1 cells. (D) Quantification of the number of NK and ILC1 cells in the lung ( n = 3–4 per group, x ‾ ± S E M ), representative of 3 experiments performed. Statistical significance was determined by one-way ANOVA and Tukey’s multiple correction (* p < 0.05). (G) Frequency of total NK1.1 + IL-12Fc + and Thy1.1 + cells at the indicated time points in the spleen and lung of IFN-γ reporter mice ( n = 3–4 per group, x ‾ ± S E M ). Statistical significance was determined by one-way ANOVA and Tukey’s multiple correction (** p < 0.005, *** p < 0.001, and **** p < 0.0001).

Article Snippet: Armenian hamster anti-mouse CD3 (clone: 145-2C11) , BioXcell , Catalog: BE0001-1; RRID: AB_1107634.

Techniques: Control, Expressing

Journal: Cell reports

Article Title: De-coupling immune parameters and toxicity associated with IL-12 agonism

doi: 10.1016/j.celrep.2025.115840

Figure Lengend Snippet: IFN-γ Thy1.1 reporter mice received daily i.p. injections of IL-12 Fc, IL-12 3x Ala Fc, or an IgG2a Fc fragment, and spleen and lung were harvested on day 7 for analysis. (A) Quantification of the total number of CD3 + (CD4 − CD8 − ) DN NKT cells in the spleen ( n = 6 group, pooled data from 2 independent experiments, x ‾ ± S E M ). Statistical significance was determined by one-way ANOVA and Tukey’s multiple correction (* p < 0.05). (B) Frequency of splenic Thy1.1 + CD3 + (CD4 − CD8 − ) DN NKT cells ( n = 6 group, pooled data from 2 independent experiments, x ‾ ± S E M ). Statistical significance was determined by one-way ANOVA and Tukey’s multiple correction (** p < 0.005 and **** p < 0.0001). (C) Representative flow plot depicting bound IL-12 agonist x Thy1.1 expression for CD3 + (CD4 − CD8 − ) DN NKT cells from the spleen and lung. (D) Impact of IL-12 Fc on the frequency of Thy1.1 + splenic CD3 + (CD4 − CD8) DN NKT cells at the indicated time points ( n = 3–4 mice/group, x ‾ ± S E M ). Statistical significance was determined by one-way ANOVA and Tukey’s multiple correction (*** p < 0.001 and **** p < 0.0001). Data are from a representative experiment of 2–3 per time point. (E) Quantification of splenic CD3 + CD4 + or CD8 + T cells on day 7. n = 5 per group. Data shown are representative of 2 experiments performed, with statistical significance determined by one-way ANOVA and Tukey’s multiple correction. (F) Representative flow plot depicting CD11aexpression x Thy1.1 expression to identify splenic effector CD4 + and CD8 + T cells. (G) Quantification of the number of splenic Thy1.1 + CD4 + or CD8 + T cells on day 7. Data are pooled from 2 independent experiments with n = 3/group, and statistical significance was determined by one-way ANOVA and Tukey’s multiple correction (* p < 0.05, ** p < 0.005, and **** p < 0.0001). (H) Representative flow plot depicting bound IL-12 agonist x Thy1.1 expression by CD4 + and CD8 + T cells from the spleen and lung on day 7.

Article Snippet: Armenian hamster anti-mouse CD3 (clone: 145-2C11) , BioXcell , Catalog: BE0001-1; RRID: AB_1107634.

Techniques: Expressing

Journal: Cell reports

Article Title: De-coupling immune parameters and toxicity associated with IL-12 agonism

doi: 10.1016/j.celrep.2025.115840

Figure Lengend Snippet: C57BL/6 (WT) mice received an i.p. injection of 200 μg/mouse of depleting antibody (Ab). After 24 h, they received daily i.p. injections of IL-12 Fc, and body weight was monitored. Mice were euthanized on day 7. All data are from 1 of 2 experiments performed with n = 3–5/group. Data are presented as x ‾ ± S E M . (A) Body weight was measured daily and statistical significance determined by one-way ANOVA and Tukey’s multiple correction. (* p < 0.05 and **** p < 0.0001). (B) Blood was collected on day 7 and IFN-γ levels measured by ELISA. Statistical significance was determined by one-way ANOVA and Tukey’s multiple correction (** p < 0.005 and **** p < 0.0001). (C) Quantification of the number of splenic CD3 − NK1.1 + NK cells, CD3 + CD4 − CD8 − DN NKT cells, and CD3 + CD4 + and CD3 + CD8 + T cells on day 7. Statistical significance was determined by one-way ANOVA and Tukey multiple correction. (* p < 0.05, ** p < 0.005, and **** p < 0.0001). (D) Representative flow plot depicting Tbetx KLRG1 expression after pre-gating on CD3 + CD4 − CD8 − DN NKT cells. Frequency and total number of Tbet + KLRG1 + CD3 + CD4 − CD8 − DN NKT cells. Statistical significance was determined by one-way ANOVA and Tukey’s multiple correction (* p < 0.05, ** p < 0.005, and **** p < 0.0001). (E) Serum levels of aspartate and alanine aminotransferase on day 7. Data presented are pooled from 2 independent experiments with n = 5–10/group. Statistical significance was determined by one-way ANOVA and Tukey’s multiple correction (* p < 0.05 and *** p < 0.001).

Article Snippet: Armenian hamster anti-mouse CD3 (clone: 145-2C11) , BioXcell , Catalog: BE0001-1; RRID: AB_1107634.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Expressing