Journal: Advanced Functional Materials

Article Title: Multifunctional Nanovaccine Sensitizes Breast Cancer to Immune Checkpoint Therapy

doi: 10.1002/adfm.202401749

Figure Lengend Snippet: Multifunctional nanovaccine synergizes with αOX40 immune checkpoint in 4T1‐bearing mice. a) Timeline (days) of tumor inoculation in BALB/c mice, immunization scheme, and immune checkpoint therapy. b) Tumor growth curve. Data are presented as mean ± s.e.m. ( N = 5 animals). p values correspond to tumor volume at day 22 after tumor inoculation relative to the PBS‐treated group and * p values are relative to the nanovaccine + αOX40 group. One‐way ANOVA followed by Tukey post‐hoc test. c) Kaplan‐Meier overall survival over time graph for mice inoculated with 7 × 10 5 4T1 cells (two independent assays were performed with N = 8 animals for NP1+NP2+αOX40 group and N = 5 animals for the remaining groups). Log‐Rank test, for the combination NP1_ɑ‐Lac+NP2+αOX40, compared to all other treatment groups ( p values), and for PBS compared to NP‐based treatment groups ( *P values). Tumor‐infiltrating immune cell populations ford) CD3 + , e) CD3 + CD4 + , f) CD3 + CD8 + , g) CD3 + CD8 + TNF‐α + , h) CD3 + CD8 + IFN‐γ + , i) CD3 + CD8 + IL‐2 + , j) CD3 + CD8 + CD107b + , k) CD3 + CD8 + PD1 + , and l) CD11b + Gr‐1 + . Tumors were isolated on day 19 after tumor cell inoculation. Quantification was performed by flow cytometric analysis. Data are presented as mean ± s.d., N = 3 animals, n = 3 measurements per experiment. One‐way ANOVA followed by Tukey post‐hoc test. p values relative to the PBS‐treated group.

Article Snippet: CD8 ± T cell panel : CD3‐PerCP‐Vio700 (Miltenyi Biotec, Cat.# 130‐109‐240, mouse, clone: REA606, 1:10 dilution), CD8a‐PE (Miltenyi Biotec, Cat.# 130‐109‐247, mouse, clone: REA601, 1:10 dilution), CD107b‐APC‐Vio770 (Miltenyi Biotec, Cat.# 130‐106‐292, mouse, clone: M3/84, 1:10 dilution), CD279 (PD‐1)‐VioBlue (Miltenyi Biotec, Cat.# 130‐102‐741, mouse, clone: HA2‐7B1, 1:10 dilution).

Techniques: Isolation

Journal: Advanced Functional Materials

Article Title: Multifunctional Nanovaccine Sensitizes Breast Cancer to Immune Checkpoint Therapy

doi: 10.1002/adfm.202401749

Figure Lengend Snippet: PD‐1 blockade did not improve the anti‐tumor effect of the dual therapy. a) Timeline (days) of tumor inoculation in BALB/c mice, immunization scheme, and immune checkpoint therapy. b) Tumor growth curve. Data are presented as mean ± s.e.m. (N = 5 animals). One‐way ANOVA followed by Tukey post‐hoc test. P values correspond to tumor volume at day 22 after tumor inoculation relative to the PBS‐treated group. c) Individual tumor volume at day 22 (N = 5 animals) with mean ± s.e.m. P values correspond to tumor volume at day 22 after tumor inoculation relative to the PBS‐treated group. One‐way ANOVA followed by Tukey post‐hoc test. Tumor‐infiltrating immune cell populations for d) CD3 + , e) CD3 + CD4 + , f) CD3 + CD8 + , g) CD3 + CD8 + CD107b + , h) CD3 + CD8 + TNF‐α + , i) CD3 + CD8 + IFN‐γ + , j) CD3 + CD8 + IL‐2 + , k) CD3 + CD8 + PD‐1 + , and l) CD11b + Gr‐1 + . Tumors were isolated on day 22 after tumor cell inoculation. Quantification was performed by flow cytometry. Data are presented as mean ± s.d., N = 3 animals, n = 3 measurements per experiment. One‐way ANOVA followed by Tukey post‐hoc test. P values relative to the PBS‐treated group.

Article Snippet: CD8 ± T cell panel : CD3‐PerCP‐Vio700 (Miltenyi Biotec, Cat.# 130‐109‐240, mouse, clone: REA606, 1:10 dilution), CD8a‐PE (Miltenyi Biotec, Cat.# 130‐109‐247, mouse, clone: REA601, 1:10 dilution), CD107b‐APC‐Vio770 (Miltenyi Biotec, Cat.# 130‐106‐292, mouse, clone: M3/84, 1:10 dilution), CD279 (PD‐1)‐VioBlue (Miltenyi Biotec, Cat.# 130‐102‐741, mouse, clone: HA2‐7B1, 1:10 dilution).

Techniques: Isolation, Flow Cytometry

Journal: Advanced Functional Materials

Article Title: Multifunctional Nanovaccine Sensitizes Breast Cancer to Immune Checkpoint Therapy

doi: 10.1002/adfm.202401749

Figure Lengend Snippet: Nanovaccines synergize with αOX40 immune checkpoint in the EO771 luminal B‐type breast cancer mouse model. a) Timeline (days) of tumor inoculation in C57BL/6J mice, immunization scheme, and immune checkpoint therapy. b) Tumor growth curve. Data are presented as mean ± s.e.m. (N = 10 animals). One‐way ANOVA followed by Tukey post‐hoc test. P values correspond to tumor volume at day 22 after tumor inoculation relative to the PBS‐treated group and *P values are relative to the nanovaccine + αOX40 group. c) Individual tumor volume at day 22 (N = 10 animals) with mean ± s.e.m. One‐way ANOVA followed by Tukey post‐hoc test. d) Kaplan‐Meier overall survival over time graph for mice inoculated with 1 × 10 6 EO771 cells (N = 6–7 animals). Log‐Rank test, for the combination NP1_KRAS (s.c.) + NP1_KRAS (i.t.) + αOX40 compared to KRAS+siTGF‐β1+Adj + αOX40 group. Tumor‐infiltrating immune cell populations for live CD45 + cells: e) CD3 + , f) CD3 + CD4 + , g) CD3 + CD8 + , h) CD3 + CD8 + CD107b + , and i) CD3 + CD4 + CD25 + CD127‐. Tumors were isolated on day 22 after tumor cell inoculation. Quantification was performed by flow cytometry. Data are presented as mean ± s.d., N = 3 animals, n = 3 measurements per experiment. One‐way ANOVA followed by Tukey post‐hoc test.

Article Snippet: CD8 ± T cell panel : CD3‐PerCP‐Vio700 (Miltenyi Biotec, Cat.# 130‐109‐240, mouse, clone: REA606, 1:10 dilution), CD8a‐PE (Miltenyi Biotec, Cat.# 130‐109‐247, mouse, clone: REA601, 1:10 dilution), CD107b‐APC‐Vio770 (Miltenyi Biotec, Cat.# 130‐106‐292, mouse, clone: M3/84, 1:10 dilution), CD279 (PD‐1)‐VioBlue (Miltenyi Biotec, Cat.# 130‐102‐741, mouse, clone: HA2‐7B1, 1:10 dilution).

Techniques: Isolation, Flow Cytometry

Journal: Advanced Functional Materials

Article Title: Multifunctional Nanovaccine Sensitizes Breast Cancer to Immune Checkpoint Therapy

doi: 10.1002/adfm.202401749

Figure Lengend Snippet: Nanovaccine and αOX40 combination downregulated CD8 + T‐cell exhaustion signature and upregulated their memory profile. a) Unsupervised clustering of the single‐cell RNA‐seq profiles of CD8 + T cells identified in 4T1 tumors. b) Bar graphs showing the number of cells present in each cluster from each treatment group for each cell cluster identified. c) Clusters distribution between each treatment group for each cell cluster identified. d) Feature plot depicting the expression distribution of selected genes within clusters. e,g) Projection of a stem‐like T‐cells signature on a t‐sne plot of the CD8 + T cells and violin plots showing up‐regulation in combination therapy compared to PBS ( p value = p value < 0.01, ANOVA and a Tukey post‐hoc test). f,h) Projection of exhaustion T‐cells signature on t‐sne plot of the CD8 + T cells and violin plots showing up‐regulation in combination therapy compared to PBS ( p value = p value < 4.76 × 10 −9 , ANOVA and a Tukey post‐hoc test). COM = NP (s.c.) + NP (i.t.) + αOX40; NP = NP (s.c.) + NP (i.t.); OX40 = αOX40.

Article Snippet: CD8 ± T cell panel : CD3‐PerCP‐Vio700 (Miltenyi Biotec, Cat.# 130‐109‐240, mouse, clone: REA606, 1:10 dilution), CD8a‐PE (Miltenyi Biotec, Cat.# 130‐109‐247, mouse, clone: REA601, 1:10 dilution), CD107b‐APC‐Vio770 (Miltenyi Biotec, Cat.# 130‐106‐292, mouse, clone: M3/84, 1:10 dilution), CD279 (PD‐1)‐VioBlue (Miltenyi Biotec, Cat.# 130‐102‐741, mouse, clone: HA2‐7B1, 1:10 dilution).

Techniques: RNA Sequencing, Expressing

Journal: PLoS ONE

Article Title: Rapid Inflammation in Mice Lacking Both SOCS1 and SOCS3 in Hematopoietic Cells

doi: 10.1371/journal.pone.0162111

Figure Lengend Snippet: ( A ) Representative flow cytometry profiles of CD8 and CD44 expression on TCRβ + T cells in spleens mice 14d after tamoxifen or vehicle treatment. ( B ) Proportions of CD8 + T cells expressing high levels of CD44 (CD44 hi ) in spleens of mice at the times indicated after tamoxifen or vehicle treatment. ( C ) Proportions of naïve (CD44 - CD62L + ), central memory (CD44 + CD62L + ) and effector memory (CD44 + CD62L - ) CD3 + T cells in spleens of mice 50d (top panels) or 180d (bottom) after tamoxifen or vehicle treatment (m, moribund). Mean ± SD is shown with * p<0.05 for comparison of γ S1S3 (moribund) with all genotypes at day 50 except γ S1 (non-moribund day 50). ** p<0.05 for comparison of γ S1 (moribund) with all genotypes at day 180 except γ S1 (non-moribund, day 180), one-way ANOVA with Tukey’s multiple comparisons test, n = 7–20 mice per group.

Article Snippet: CD8 + cells were enriched from spleen cell preparations via positive selection using anti-CD8a (Ly-2) microbeads (Miltenyi Biotech) and then CD45.2 + CD3 + CD8 + T cells were sorted by flow cytometry.

Techniques: Flow Cytometry, Expressing, Comparison

Journal: PLoS ONE

Article Title: Rapid Inflammation in Mice Lacking Both SOCS1 and SOCS3 in Hematopoietic Cells

doi: 10.1371/journal.pone.0162111

Figure Lengend Snippet: ( A ) Representative flow cytometry profiles showing expression of activation and homing markers as indicated on CD8 + CD44 high cells from spleens of mice 14d after tamoxifen or vehicle treatment. ( B ) CD8 + T cells were purified from spleens of mice 14d following tamoxifen or vehicle treatment. The cells were stimulated for 72 hours in culture in the presence of IL-15 or IL-2 plus anti-CD3/CD28. Expression of CD25, CD69, CD44 and CD62L before and after culture in a representative experiment is shown.

Article Snippet: CD8 + cells were enriched from spleen cell preparations via positive selection using anti-CD8a (Ly-2) microbeads (Miltenyi Biotech) and then CD45.2 + CD3 + CD8 + T cells were sorted by flow cytometry.

Techniques: Flow Cytometry, Expressing, Activation Assay, Purification

Journal: PLoS ONE

Article Title: Rapid Inflammation in Mice Lacking Both SOCS1 and SOCS3 in Hematopoietic Cells

doi: 10.1371/journal.pone.0162111

Figure Lengend Snippet: Concentrations of cytokines/chemokines in supernatant of CD8 + T cells 48hr after stimulation with IL-2 plus anti-CD3/CD28 ( A ) or IL-15 ( B ). Mean ± SD is shown. CD45.2 + CD3 + CD8 + T cells were flow-sorted from spleens of mice 14d after tamoxifen or vehicle treatment. Mean ± SD is shown with * p<0.05 for comparison of γ S1S3 with TAM- and S3 . ** p<0.05 for comparison of γ S1 with TAM- and S3 , one-way ANOVA with Tukey’s multiple comparisons test, n = 6–7 mice per group.

Article Snippet: CD8 + cells were enriched from spleen cell preparations via positive selection using anti-CD8a (Ly-2) microbeads (Miltenyi Biotech) and then CD45.2 + CD3 + CD8 + T cells were sorted by flow cytometry.

Techniques: Comparison