Journal: Molecular cancer

Article Title: Biologically targeted dual adaptive and innate nano-Immunotherapy for clear cell renal cell carcinoma treatment.

doi: 10.1186/s12943-025-02382-y

Figure Lengend Snippet: Fig. 2 The MINP platform mimics plate-bound agonistic antibodies to crosslink and activate 4-1BB and/or CD27 receptors on mouse CD8+ T cells and NK cells in vitro. (A) Frequencies of CD69+ activated mouse CD8+ T cells after incubation for 6 h with different formulations of α-CA-9, α-4-1BB, and/or α-CD27 (n = 4). (B) Frequencies of CD69+ activated mouse NKp46+ NK cells after incubation for 6 h with different formulations of α-CA-9, α-4-1BB, and/ or α-CD27 (n = 4). (C) Frequencies of IFN-γ+ activated mouse CD8+ T cells after incubation for 72 h with different formulations of α-CA-9, α-4-1BB, and/or α-CD27 (n = 4). (D) Frequencies of IFN-γ+ activated mouse NK cells after incubation for 72 h with different formulations of α-CA-9, α-4-1BB, and/or α-CD27 (n = 4). Anti-human CA-9, anti-mouse 4-1BB, and CD27 agonistic antibodies were used in these studies. Data are presented as mean ± SEM. All p-values were analyzed using two-way ANOVA with Tukey’s HSD multiple comparisons post-hoc test

Article Snippet: The actual degree of DBCOfunctionalization was calculated via UV-visible absorption spectroscopy using a DBCO absorption coefficient of 12,000 M− 1 L cm− 1 at 310 nm, an immunoglobulin absorption coefficient of 1.33 mg/mL cm− 1 at 280 nm for the anti-mouse 4-1BB and anti-mouse CD27 antibodies (as provided by BioXCell), 1.45 mg/mL cm− 1 for Girentuximab (anti-human CA-9), 1.66 mg/mL cm− 1 for Utomilumab (anti-human 4-1BB), and 1.61 mg/mL cm− 1 for Varlilumab (anti-human CD27), with a correction factor of 1.089 at 280 nm.

Techniques: In Vitro, Incubation

Journal: Molecular cancer

Article Title: Biologically targeted dual adaptive and innate nano-Immunotherapy for clear cell renal cell carcinoma treatment.

doi: 10.1186/s12943-025-02382-y

Figure Lengend Snippet: Fig. 3 Human CA-9-targeted chimeric MINPs mediate mouse CD8+ T cells and NK cells to kill hCA-9-expressing RCC cells in vitro. (A) In vitro lysis of free antibody- and chimeric MINP-pretreated calcein-loaded RCC cells after coculture with CD8+ T cells for 6 h. The effector cell-to-target (E: T) ratio was 5:1. (B) In vitro lysis of free antibody- and chimeric MINP-pretreated calcein-loaded RCC cells after coculturing for 6 h with mouse NK cells. The E: T ratio was 5:1. (C) Relative viability of free antibody- and chimeric MINP-pretreated RCC cells after coculture for 72 h with mouse CD8+ T cells, as determined through an MTS assay (E: T ratio = 5:1). (D) Relative viability of free antibody- and chimeric MINP-pretreated RCC cells after coculture for 72 h with mouse CD8+ T cells, as determined through an MTS assay (E: T ratio = 5:1). The chimeric MINPs were functionalized with anti-human CA-9, anti-mouse 4-1BB, and/or CD27 agonistic antibodies in these in vitro studies. (E) Representative CLSM images of free antibody- and chimeric MINP-pretreated calcein-loaded RAG/hCA-9 cells after coculture for 6 h with CD8+ T cells or NK cells. Calcein-free RAG/hCA-9 cells and calcein-loaded RAG/hCA-9 cells were used as negative and positive controls, respectively, for the calcein channel. The E: T ratio was 5:1. The merge channel images are the same as those shown in Fig. S12. Data are presented as mean ± SEM. All p-values were analyzed using two-way ANOVA with Tukey’s HSD multiple comparisons post-hoc test

Article Snippet: The actual degree of DBCOfunctionalization was calculated via UV-visible absorption spectroscopy using a DBCO absorption coefficient of 12,000 M− 1 L cm− 1 at 310 nm, an immunoglobulin absorption coefficient of 1.33 mg/mL cm− 1 at 280 nm for the anti-mouse 4-1BB and anti-mouse CD27 antibodies (as provided by BioXCell), 1.45 mg/mL cm− 1 for Girentuximab (anti-human CA-9), 1.66 mg/mL cm− 1 for Utomilumab (anti-human 4-1BB), and 1.61 mg/mL cm− 1 for Varlilumab (anti-human CD27), with a correction factor of 1.089 at 280 nm.

Techniques: Expressing, In Vitro, Lysis, MTS Assay

Journal: Molecular cancer

Article Title: Biologically targeted dual adaptive and innate nano-Immunotherapy for clear cell renal cell carcinoma treatment.

doi: 10.1186/s12943-025-02382-y

Figure Lengend Snippet: Fig. 6 Human CA-9-targeted chimeric MINPs inhibit RAG/hCA-9 tumor growth by activating tumor-infiltrating CD8+ T cells and NK cells in immunocom petent mice. (A) Ex vivo fluorescence images of RAG/hCA-9 tumors preserved 48 h after i.v. administration of Cy5-labeled α-CA-9 NPs plus Cy5-labeled α-4-1BB/α-CD27 NPs, or Cy5-labeled α-CA-9/α-4-1BB/α-CD27 NPs (n = 5, except for the non-treatment control group where n = 3). (B) Frequencies of active IFN-γ+ tumor-infiltrating NK cells. (C) Frequencies of active IFN-γ+ tumor-infiltrating CD8+ T cells. (D) frequencies of tumor-infiltrating TEM cells. (E) The TEM-to-TCM ratio. The phenotypes of tumor-infiltrating lymphocytes were determined by the FACS method 4 days after 2 targeted or non-targeted MINP treatments (n = 5). (F) Representative immunohistochemistry images of RAG/hCA-9 tumors after treatment with free combinational antibodies or hCA-9-targeted MINPs. The chimeric MINPs were functionalized using anti-human CA-9, anti-mouse 4-1BB, and/or CD27 agonistic antibodies in these in vivo studies. Data are presented as mean ± SEM. All p-values were analyzed using two-way ANOVA with Tukey’s HSD multiple comparisons post-hoc test

Article Snippet: The actual degree of DBCOfunctionalization was calculated via UV-visible absorption spectroscopy using a DBCO absorption coefficient of 12,000 M− 1 L cm− 1 at 310 nm, an immunoglobulin absorption coefficient of 1.33 mg/mL cm− 1 at 280 nm for the anti-mouse 4-1BB and anti-mouse CD27 antibodies (as provided by BioXCell), 1.45 mg/mL cm− 1 for Girentuximab (anti-human CA-9), 1.66 mg/mL cm− 1 for Utomilumab (anti-human 4-1BB), and 1.61 mg/mL cm− 1 for Varlilumab (anti-human CD27), with a correction factor of 1.089 at 280 nm.

Techniques: Ex Vivo, Fluorescence, Labeling, Control, Immunohistochemistry, In Vivo

Journal: Molecular cancer

Article Title: Biologically targeted dual adaptive and innate nano-Immunotherapy for clear cell renal cell carcinoma treatment.

doi: 10.1186/s12943-025-02382-y

Figure Lengend Snippet: Fig. 8 The MINP platform reduces hepatotoxicity induced by agonistic α-4-1BB and α-CD25 antibodies in healthy immunocompetent mice. (A) Serum ALT, AST, and BUN levels recorded for healthy BALB/c mice 48 h after 4 i.v. administrations of free α-CA-9, α-4-1BB, and α-CD27, or α-CA-9/α-4-1BB/α- CD27 NPs (chimeric MINPs) (n = 7, except for mice treated with MINPs, where n = 8). (B) Representative H&E-stained images of the liver, lung, spleen, and kidney preserved from mice at the study endpoint. (C) Representative immunohistochemistry images of liver sections preserved at the study endpoint after different treatments. (D) Serum DyLight 650 fluorescent intensity 6 h after i.v. administration of DyLight 650 labeled α-CA-9, α-4-1BB, and α-CD27 or α-CA-9/α-4-1BB/α-CD27 NPs (functionalized with DyLight 650-labeled antibodies) (n = 5, except for the non-treatment control group, where n = 3). The chimeric MINPs were functionalized using anti-human CA-9, anti-mouse 4-1BB, and CD27 agonistic antibodies in these in vivo studies. Data are presented as mean ± SEM. All p-values were analyzed using two-way ANOVA with Tukey’s HSD multiple comparisons post-hoc test

Article Snippet: The actual degree of DBCOfunctionalization was calculated via UV-visible absorption spectroscopy using a DBCO absorption coefficient of 12,000 M− 1 L cm− 1 at 310 nm, an immunoglobulin absorption coefficient of 1.33 mg/mL cm− 1 at 280 nm for the anti-mouse 4-1BB and anti-mouse CD27 antibodies (as provided by BioXCell), 1.45 mg/mL cm− 1 for Girentuximab (anti-human CA-9), 1.66 mg/mL cm− 1 for Utomilumab (anti-human 4-1BB), and 1.61 mg/mL cm− 1 for Varlilumab (anti-human CD27), with a correction factor of 1.089 at 280 nm.

Techniques: Staining, Immunohistochemistry, Labeling, Control, In Vivo

Journal: Science Advances

Article Title: Small molecules from antibody pharmacophores (SMAbPs) as a hit identification workflow for immune checkpoints

doi: 10.1126/sciadv.adq5540

Figure Lengend Snippet: ( A ) Chemical structure of MG-C-30. ( B ) Activation of natural killer cells from PBMCs of healthy donors as revealed by the frequency of CD69 + cells in the presence of anti-hCD27 mAb (250 nM) and multiple concentrations of MG-C-30. Error bars represent SD ( n = 5). ( C ) Tumor volume in the EG7-OVA mouse model of vehicle control, MG-C-30 (25 or 50 mg/kg, po), and anti-mCD27 mAbs groups of C57BL/6 mice. Error bars represent SEM ( n = 8). ns denotes nonsignificant; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 relative to control.

Article Snippet: When the average tumor volume reached ~50 mm 3 , the mice were randomly assigned to four groups of eight mice each: control group, 25 mg/kg (po) of MG-C-30 treatment group, 50 mg/kg (po) of MG-C-30 treatment group, and anti-mCD27 mAb group (10 mg/kg; Bio X Cell, BE0348).

Techniques: Activation Assay, Control