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hek blue il 12 reporter cell line 24 (InvivoGen)

Name: hek blue il 12 reporter cell line 24
Brand: InvivoGen
Rating: 94 (72 reviews)

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InvivoGen hek blue il 12 reporter cell line 24
Hek Blue Il 12 Reporter Cell Line 24, supplied by InvivoGen, used in various techniques. Bioz Stars score: 94/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 72 article reviews

hek blue il 12 reporter cell line 24 - by Bioz Stars, 2026-03

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a , Schematic of LbL-NP assembly with either Mal or Ni linker chemistries for the conjugation of IL-12. b , Quantification of total PLE and PLR retained with LbL NPs following incubation in cell-free ascites fluid at 37 °C (* indicates a fluorophore-tagged polymer; n = 2–6 independent release assays; mean ± s.d.). c , Quantification of the total IL-12 available for monoclonal antibody binding from Mal NPs incubated in either diH 2 O or 10% FBS media ( n = 2 independent batches with two technical replicates; mean ± s.d.). NS, not significant. d , Quantification of the total IL-12 released from LbL NPs on incubation in cell-free ascites fluid at 37 °C ( n = 3–6 independent release assays; mean ± s.d.). e , Representative flow cytometry fluorescence histogram of HM-1 cells incubated with UL or PLE-coated LbL NPs for 4 h in vitro. f , Quantification of median fluorescence intensity (MFI) of treated HM-1 cells shown in e ( n = 4 technical replicates; mean ± s.d.). g , h , Representative confocal images of HM-1 cells incubated with UL ( g ) or LbL ( h ) NPs for 4 h—UL images adjusted relative to LbL to visualize internalized NPs (blue, Hoechst 33342 nuclear stain; green, WGA cell surface stain; cyan, NPs). Data are representative of at least two independent experiments. Statistical comparisons in c and f were performed using two- and one-way analysis of variance (ANOVA), respectively, with Tukey’s multiple comparisons test.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a , Schematic of LbL-NP assembly with either Mal or Ni linker chemistries for the conjugation of IL-12. b , Quantification of total PLE and PLR retained with LbL NPs following incubation in cell-free ascites fluid at 37 °C (* indicates a fluorophore-tagged polymer; n = 2–6 independent release assays; mean ± s.d.). c , Quantification of the total IL-12 available for monoclonal antibody binding from Mal NPs incubated in either diH 2 O or 10% FBS media ( n = 2 independent batches with two technical replicates; mean ± s.d.). NS, not significant. d , Quantification of the total IL-12 released from LbL NPs on incubation in cell-free ascites fluid at 37 °C ( n = 3–6 independent release assays; mean ± s.d.). e , Representative flow cytometry fluorescence histogram of HM-1 cells incubated with UL or PLE-coated LbL NPs for 4 h in vitro. f , Quantification of median fluorescence intensity (MFI) of treated HM-1 cells shown in e ( n = 4 technical replicates; mean ± s.d.). g , h , Representative confocal images of HM-1 cells incubated with UL ( g ) or LbL ( h ) NPs for 4 h—UL images adjusted relative to LbL to visualize internalized NPs (blue, Hoechst 33342 nuclear stain; green, WGA cell surface stain; cyan, NPs). Data are representative of at least two independent experiments. Statistical comparisons in c and f were performed using two- and one-way analysis of variance (ANOVA), respectively, with Tukey’s multiple comparisons test.

Article Snippet: HEK-Blue IL-12 and HEK-Blue IL-2 (InvivoGen) cells were cultured and used for IL-12 and IL-15sa bioactivity assessment, respectively, according to the manufacturer’s instructions.

Techniques: Conjugation Assay, Incubation, Polymer, Binding Assay, Flow Cytometry, Fluorescence, In Vitro, Staining

a , Chemical structure of headgroup-modified lipids with either chelated nickel or N-aryl maleimide. N-aryl maleimide was employed to prevent potential thiosuccinimide retro-Michael addition and subsequent thiol-exchange, as this headgroup favors thiosuccinimide hydrolysis into stable succinamic acid thioethers. b , IL-12 crystal structure derived from PDB 1F45 showing C-terminus used to engineered terminal poly-histidine (polyHis) tag and terminal cysteine. c , Reaction pathway for irreversible maleimide-based conjugation with thiols.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a , Chemical structure of headgroup-modified lipids with either chelated nickel or N-aryl maleimide. N-aryl maleimide was employed to prevent potential thiosuccinimide retro-Michael addition and subsequent thiol-exchange, as this headgroup favors thiosuccinimide hydrolysis into stable succinamic acid thioethers. b , IL-12 crystal structure derived from PDB 1F45 showing C-terminus used to engineered terminal poly-histidine (polyHis) tag and terminal cysteine. c , Reaction pathway for irreversible maleimide-based conjugation with thiols.

Techniques: Modification, Derivative Assay, Conjugation Assay

a , Intensity-weighted hydrodynamic size (Z-avg), number average size (#-avg), and polydispersity index (PDI) of NPs during synthesis as measured via dynamic light scattering (mean ± s.d.). b , Zeta potential of NPs during synthesis as measured via electrophoretic mobility in deionized water (mean ± s.d.). c , Yield and weight loading of IL-12 for unlayered and layered particles with nickel-histidine linker (Ni-UL and Ni-LbL) and unlayered and layered particles with a maleimide-cysteine bond (Mal-UL and Mal-LbL) (mean ± s.e.m.). d-e , Negative-stain (NS) transmission electron microscopy (TEM) with phosphotungstic acid of particles during synthesis with nickel-containing lipids ( d ) and maleimide-containing lipids ( e ) - scale bars represent 200 nm. (f) Unlayered liposomal NPs devoid of IL-12 (UL-NP) or maleimide-cysteine IL-12 conjugated NPs (Mal-UL) were deposited on TEM copper grids followed by binding to either anti-IL-12 or isotype IgG antibody (Ab) and then exposed to 1 nm gold-labeled secondary anti-IgG FAbs. Shown are control UL-NPs without IL-12 conjugation exposed to anti-IL12 Ab (i), Mal-UL NPs incubated with isotype control IgG (ii) or Mal-UL NPs exposed to anti-IL-12 Ab (iii, iv, v). Scale bars 100 nm. Data are presented as mean values ± error with at least n = 3 independent batches of NPs. Statistical comparisons in c were performed using two-way analysis of variance (ANOVA) with Tukey’s multiple-comparisons test.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a , Intensity-weighted hydrodynamic size (Z-avg), number average size (#-avg), and polydispersity index (PDI) of NPs during synthesis as measured via dynamic light scattering (mean ± s.d.). b , Zeta potential of NPs during synthesis as measured via electrophoretic mobility in deionized water (mean ± s.d.). c , Yield and weight loading of IL-12 for unlayered and layered particles with nickel-histidine linker (Ni-UL and Ni-LbL) and unlayered and layered particles with a maleimide-cysteine bond (Mal-UL and Mal-LbL) (mean ± s.e.m.). d-e , Negative-stain (NS) transmission electron microscopy (TEM) with phosphotungstic acid of particles during synthesis with nickel-containing lipids ( d ) and maleimide-containing lipids ( e ) - scale bars represent 200 nm. (f) Unlayered liposomal NPs devoid of IL-12 (UL-NP) or maleimide-cysteine IL-12 conjugated NPs (Mal-UL) were deposited on TEM copper grids followed by binding to either anti-IL-12 or isotype IgG antibody (Ab) and then exposed to 1 nm gold-labeled secondary anti-IgG FAbs. Shown are control UL-NPs without IL-12 conjugation exposed to anti-IL12 Ab (i), Mal-UL NPs incubated with isotype control IgG (ii) or Mal-UL NPs exposed to anti-IL-12 Ab (iii, iv, v). Scale bars 100 nm. Data are presented as mean values ± error with at least n = 3 independent batches of NPs. Statistical comparisons in c were performed using two-way analysis of variance (ANOVA) with Tukey’s multiple-comparisons test.

Techniques: Zeta Potential Analyzer, Staining, Transmission Assay, Electron Microscopy, Binding Assay, Labeling, Control, Conjugation Assay, Incubation

a – c , B6C3F1 mice ( n = 8 animals per group for 0–24 h and n = 3 animals per group for 24–96 h) inoculated with 10 6 HM-1-luc tumour cells on day 0 were administered fluorescently tagged NPs carrying 20 µg of IL-12 (or an equivalent dose of free IL-12) on day 14. Whole-animal imaging NP fluorescence ( a ) and IL-12 fluorescence for UL ( b ) and LbL ( c ) NP treatments are shown from the i.p. space collected over time post-dosing (mean ± s.d.). d , e , B6C3F1 mice ( n = 4 animals per group) inoculated with 10 6 HM-1-luc tumour cells on day 0 were administered 130 µg of lipids in fluorescently tagged versions of LbL NPs or UL NPs (devoid of IL-12) on day 14. UGT and omentum tissues were harvested at 1, 2, 4, 12 and 24 h after dosing and imaged ex vivo via IVIS. Weight-normalized tissue NP fluorescence in UGT ( d ) and omentum ( e ) are shown (mean ± s.d.). f – h , B6C3F1 mice ( n = 4 animals per group) were treated as that in a – c ( f – h , respectively). Four hours after dosing, animals were euthanized, and tissues were analysed ex vivo via IVIS. Weight-normalized tissue NP fluorescence (mean ± s.d.; f ) are shown, along with Pearson’s correlation coefficient for groups (eight tissues with four replicates per group) with significant ( P < 0.05) correlation between weight-normalized tissue NP fluorescence and BLI 4 h after dosing ( g ), and representative omentum and UGT tissue IVIS BLI and NP fluorescence images for LbL NPs and UL NPs ( h ). Statistics derived from using all n from experiment with each animal as a data point. For g , the correlation significance is performed based on a two-sided t -test analysis with the null hypothesis of no ( r = 0) correlation and no adjustments for multiple comparisons. Group statistical comparisons in f were performed using a two-way ANOVA with Tukey’s multiple comparisons test.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a – c , B6C3F1 mice ( n = 8 animals per group for 0–24 h and n = 3 animals per group for 24–96 h) inoculated with 10 6 HM-1-luc tumour cells on day 0 were administered fluorescently tagged NPs carrying 20 µg of IL-12 (or an equivalent dose of free IL-12) on day 14. Whole-animal imaging NP fluorescence ( a ) and IL-12 fluorescence for UL ( b ) and LbL ( c ) NP treatments are shown from the i.p. space collected over time post-dosing (mean ± s.d.). d , e , B6C3F1 mice ( n = 4 animals per group) inoculated with 10 6 HM-1-luc tumour cells on day 0 were administered 130 µg of lipids in fluorescently tagged versions of LbL NPs or UL NPs (devoid of IL-12) on day 14. UGT and omentum tissues were harvested at 1, 2, 4, 12 and 24 h after dosing and imaged ex vivo via IVIS. Weight-normalized tissue NP fluorescence in UGT ( d ) and omentum ( e ) are shown (mean ± s.d.). f – h , B6C3F1 mice ( n = 4 animals per group) were treated as that in a – c ( f – h , respectively). Four hours after dosing, animals were euthanized, and tissues were analysed ex vivo via IVIS. Weight-normalized tissue NP fluorescence (mean ± s.d.; f ) are shown, along with Pearson’s correlation coefficient for groups (eight tissues with four replicates per group) with significant ( P < 0.05) correlation between weight-normalized tissue NP fluorescence and BLI 4 h after dosing ( g ), and representative omentum and UGT tissue IVIS BLI and NP fluorescence images for LbL NPs and UL NPs ( h ). Statistics derived from using all n from experiment with each animal as a data point. For g , the correlation significance is performed based on a two-sided t -test analysis with the null hypothesis of no ( r = 0) correlation and no adjustments for multiple comparisons. Group statistical comparisons in f were performed using a two-way ANOVA with Tukey’s multiple comparisons test.

Techniques: Imaging, Fluorescence, Ex Vivo, Derivative Assay

a-d , B6C3F1 mice ( n = 4-5 animals/group/timepoint) inoculated with 10 6 HM-1-luc tumor cells on day 0 were administered NPs carrying 20 µg IL-12 (or an equivalent dose of free IL-12) on day 14. Four or 24 hours after dosing, mice were euthanized and ascites was extracted with a syringe. Shown are the quantification of ascites IL-12 concentration four ( a , mean ± s.d.) and 24 hrs ( b , mean ± s.d.) after dosing, and quantification of IFN-γ four ( c , mean ± s.d.) and 24 hrs after dosing ( d , mean ± s.d.). e , Both healthy B6C3F1 (n = 4/group) and B6C3F1 mice (n = 4/group) inoculated with 10 6 HM-1-luc tumor cells on day 0 were administered 20 µg fluorescently-tagged LbL-NPs or UL-NPs on day 14. Tissue was harvested at 4 hrs after dosing and imaged ex-vivo via IVIS. Shown are weight-normalized recovered NP fluorescence ( c , mean ± s.d.). Each data point represents one animal in the experiment.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a-d , B6C3F1 mice ( n = 4-5 animals/group/timepoint) inoculated with 10 6 HM-1-luc tumor cells on day 0 were administered NPs carrying 20 µg IL-12 (or an equivalent dose of free IL-12) on day 14. Four or 24 hours after dosing, mice were euthanized and ascites was extracted with a syringe. Shown are the quantification of ascites IL-12 concentration four ( a , mean ± s.d.) and 24 hrs ( b , mean ± s.d.) after dosing, and quantification of IFN-γ four ( c , mean ± s.d.) and 24 hrs after dosing ( d , mean ± s.d.). e , Both healthy B6C3F1 (n = 4/group) and B6C3F1 mice (n = 4/group) inoculated with 10 6 HM-1-luc tumor cells on day 0 were administered 20 µg fluorescently-tagged LbL-NPs or UL-NPs on day 14. Tissue was harvested at 4 hrs after dosing and imaged ex-vivo via IVIS. Shown are weight-normalized recovered NP fluorescence ( c , mean ± s.d.). Each data point represents one animal in the experiment.

Techniques: Concentration Assay, Ex Vivo, Fluorescence

a – e , B6C3F1 mice ( n = 4–5 animals per group) inoculated with 10 6 HM-1-luc tumour cells on day 0 were administered fluorescently tagged NPs carrying 20 µg of IL-12 on day 14. Four hours or 1 day after dosing, the animals were euthanized, and tissues were analysed ex vivo via IVIS. Pearson’s correlation coefficient is shown for groups (eight tissues with five replicates per group) with significant ( P < 0.05) correlation between weight-normalized tissue NP fluorescence and IL-12 fluorescence 4 h after dosing ( a ), weight-normalized tissue IL-12 fluorescence 1 day after dosing in UGT and omentum (mean ± s.d.; b ), Pearson’s correlation coefficient for groups (eight tissues with five replicates per group) with significant ( P < 0.05) correlation between weight-normalized tissue IL-12 fluorescence and BLI 1 day after dosing ( c ), representative omentum and UGT tissue IVIS BLI and IL-12 fluorescence images for Mal UL and Mal LbL ( d ) and pixel-by-pixel Spearman’s correlation coefficient between IL-12 fluorescence and BLI 1 day after dosing from IVIS images across all tissues ( e ; mean ± s.d., eight tissues with five replicates per group). f , g , B6C3F1 mice were treated as that in a . One day after dosing, Mal LbL NP animals were euthanized, and the omentum containing tumour nodules was frozen in OCT compound and then frozen, sectioned and stained for confocal microscopy analysis. Representative confocal microscopy images of tumour nodules in omental tissue are shown at low ( f ) and high ( g ) magnifications. The green arrows indicate areas with a high NP signal relative to IL-12, whereas yellow arrowheads indicate areas with high IL-12 relative to NPs. Statistics derived using all n from the experiment with each animal as a data point. For a and c , correlation significance performed based on a two-sided t -test analysis with the null hypothesis of no ( r = 0) correlation and no adjustments for multiple comparisons. Group statistical comparisons were performed using two-way ANOVA for b and one-way ANOVA for e with Tukey’s multiple comparisons test.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a – e , B6C3F1 mice ( n = 4–5 animals per group) inoculated with 10 6 HM-1-luc tumour cells on day 0 were administered fluorescently tagged NPs carrying 20 µg of IL-12 on day 14. Four hours or 1 day after dosing, the animals were euthanized, and tissues were analysed ex vivo via IVIS. Pearson’s correlation coefficient is shown for groups (eight tissues with five replicates per group) with significant ( P < 0.05) correlation between weight-normalized tissue NP fluorescence and IL-12 fluorescence 4 h after dosing ( a ), weight-normalized tissue IL-12 fluorescence 1 day after dosing in UGT and omentum (mean ± s.d.; b ), Pearson’s correlation coefficient for groups (eight tissues with five replicates per group) with significant ( P < 0.05) correlation between weight-normalized tissue IL-12 fluorescence and BLI 1 day after dosing ( c ), representative omentum and UGT tissue IVIS BLI and IL-12 fluorescence images for Mal UL and Mal LbL ( d ) and pixel-by-pixel Spearman’s correlation coefficient between IL-12 fluorescence and BLI 1 day after dosing from IVIS images across all tissues ( e ; mean ± s.d., eight tissues with five replicates per group). f , g , B6C3F1 mice were treated as that in a . One day after dosing, Mal LbL NP animals were euthanized, and the omentum containing tumour nodules was frozen in OCT compound and then frozen, sectioned and stained for confocal microscopy analysis. Representative confocal microscopy images of tumour nodules in omental tissue are shown at low ( f ) and high ( g ) magnifications. The green arrows indicate areas with a high NP signal relative to IL-12, whereas yellow arrowheads indicate areas with high IL-12 relative to NPs. Statistics derived using all n from the experiment with each animal as a data point. For a and c , correlation significance performed based on a two-sided t -test analysis with the null hypothesis of no ( r = 0) correlation and no adjustments for multiple comparisons. Group statistical comparisons were performed using two-way ANOVA for b and one-way ANOVA for e with Tukey’s multiple comparisons test.

Techniques: Ex Vivo, Fluorescence, Staining, Confocal Microscopy, Derivative Assay

a – d , B6C3F1 mice (one cohort of n = 10 animals per group) inoculated with 10 6 HM-1-luc tumour cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to NPs. Experimental timeline ( a ), IVIS whole-animal i.p. BLI readings (mean ± s.d.; b ) and overall survival ( c ). On day 30, PBMCs of surviving and naive mice ( n = 5 animals) were analysed via IFN-γ ELISpot restimulated with HM-1-luc tumour cells. Quantitation of spots detected (mean ± s.d.; d ) are shown. e – k , B6C3F1 mice inoculated with 10 6 HM-1 tumour cells on day 0 were treated on day 10 with 20 µg of IL-12 as a free cytokine or conjugated to Mal NPs (UL and LbL). Two days after dosing, ascites ( n = 6 per group) and i.p. tumour nodules (primarily omentum tissue, n = 4 per group) were harvested and processed for flow cytometry analysis. Timeline for experiment ( e ), representative flow plots of T cell (CD45 + CD3 + ) in ascites fluid ( f ), quantitation of T cells in ascites fluid ( g ), quantitation of CD8 + to CD4 + T cell ratio in ascites fluid ( h ), representative flow plots of T cell (CD45 + CD3 + ) in tumour nodules ( i ), quantitation of T cells in tumour nodules ( j ) and quantitation of CD8 + to CD4 + T cell ratio in tumour nodules ( k ). Statistics derived using all n from experiment with each animal as a data point. P values were determined by the log-rank (Mantel–Cox) test ( c ) and one-way ANOVA followed by Tukey’s multiple comparison test ( d , g , h , j and k ).

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a – d , B6C3F1 mice (one cohort of n = 10 animals per group) inoculated with 10 6 HM-1-luc tumour cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to NPs. Experimental timeline ( a ), IVIS whole-animal i.p. BLI readings (mean ± s.d.; b ) and overall survival ( c ). On day 30, PBMCs of surviving and naive mice ( n = 5 animals) were analysed via IFN-γ ELISpot restimulated with HM-1-luc tumour cells. Quantitation of spots detected (mean ± s.d.; d ) are shown. e – k , B6C3F1 mice inoculated with 10 6 HM-1 tumour cells on day 0 were treated on day 10 with 20 µg of IL-12 as a free cytokine or conjugated to Mal NPs (UL and LbL). Two days after dosing, ascites ( n = 6 per group) and i.p. tumour nodules (primarily omentum tissue, n = 4 per group) were harvested and processed for flow cytometry analysis. Timeline for experiment ( e ), representative flow plots of T cell (CD45 + CD3 + ) in ascites fluid ( f ), quantitation of T cells in ascites fluid ( g ), quantitation of CD8 + to CD4 + T cell ratio in ascites fluid ( h ), representative flow plots of T cell (CD45 + CD3 + ) in tumour nodules ( i ), quantitation of T cells in tumour nodules ( j ) and quantitation of CD8 + to CD4 + T cell ratio in tumour nodules ( k ). Statistics derived using all n from experiment with each animal as a data point. P values were determined by the log-rank (Mantel–Cox) test ( c ) and one-way ANOVA followed by Tukey’s multiple comparison test ( d , g , h , j and k ).

Techniques: Enzyme-linked Immunospot, Quantitation Assay, Flow Cytometry, Derivative Assay, Comparison

a , healthy B6C3F1 mice were treated on days 0 and 7 with 20 µg of IL-12 as a free cytokine or conjugated to Mal LbL-NPs. Shown are the percentage change of body weight. b-c , B6C3F1 mice (1 cohort n = 5 animals/group) inoculated with 10 6 HM-1-luc tumor cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal-LbL NPs. Serum was collected from mice one day before dosing as well as 24, 48 and 72 hrs after dosing. Shown are quantitation of serum IL-12 ( b ) and serum IFN-γ ( c ). d-h , B6C3F1 mice inoculated with 10 6 HM-1 tumor cells on day 0 were treated on days 10 with 20 µg of IL-12 as a free cytokine or conjugated to Mal NPs (UL and LbL). Two days after dosing blood (n = 5-6/group) and spleens (n = 4/group) were harvested and sent for a complete blood panel or processed for flow cytometry analysis, respectively. Shown are serum levels of liver damage markers (alanine transaminase – ALT - and aspartate aminotransferase - AST) compared to healthy mice controls ( n = 4, d ), complete blood count panel ( e ), quantitation of live leukocyte (CD45 + ) counts in spleen ( f ), and percentage of macrophage ( g ), NK ( h ) cell in splenocytes. Statistical comparisons performed using two-way ( d, e ) or one-way ( f, g, h ) analysis of variance (ANOVA) with Tukey’s multiple-comparisons (liver enzyme measurement was compared to healthy controls). In all panels, each point corresponds to a single animal, with values shown as mean ± standard deviation.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a , healthy B6C3F1 mice were treated on days 0 and 7 with 20 µg of IL-12 as a free cytokine or conjugated to Mal LbL-NPs. Shown are the percentage change of body weight. b-c , B6C3F1 mice (1 cohort n = 5 animals/group) inoculated with 10 6 HM-1-luc tumor cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal-LbL NPs. Serum was collected from mice one day before dosing as well as 24, 48 and 72 hrs after dosing. Shown are quantitation of serum IL-12 ( b ) and serum IFN-γ ( c ). d-h , B6C3F1 mice inoculated with 10 6 HM-1 tumor cells on day 0 were treated on days 10 with 20 µg of IL-12 as a free cytokine or conjugated to Mal NPs (UL and LbL). Two days after dosing blood (n = 5-6/group) and spleens (n = 4/group) were harvested and sent for a complete blood panel or processed for flow cytometry analysis, respectively. Shown are serum levels of liver damage markers (alanine transaminase – ALT - and aspartate aminotransferase - AST) compared to healthy mice controls ( n = 4, d ), complete blood count panel ( e ), quantitation of live leukocyte (CD45 + ) counts in spleen ( f ), and percentage of macrophage ( g ), NK ( h ) cell in splenocytes. Statistical comparisons performed using two-way ( d, e ) or one-way ( f, g, h ) analysis of variance (ANOVA) with Tukey’s multiple-comparisons (liver enzyme measurement was compared to healthy controls). In all panels, each point corresponds to a single animal, with values shown as mean ± standard deviation.

Techniques: Quantitation Assay, Flow Cytometry, Standard Deviation

a-j , B6C3F1 mice inoculated with 10 6 HM-1 tumor cells on day 0 were treated on days 10 with 20 µg of IL-12 as a free cytokine or conjugated to Mal NPs (UL and LbL). Two days after dosing ascites ( n = 6 animals/group) and i.p. tumor nodules (primarly omentum tissue, n = 4 animals/group) were harvested and processed for flow cytometry analysis. Shown are total counts of M1-like and M2-like macrophages (a) , logarithmic of M1-like to M2-like macrophages ratio ( b ), and total counts of PMN-MDSC ( c ), M-MDSC ( d) , and NK cells ( e ) in ascites fluid. Also shown are total counts of PMN-MDSC ( f ) and M-MDSC ( g ), logarithmic of M1-like to M2-like macrophages ratio ( h ), and total counts of M1-like and M2-like macrophages ( i ), and NK cells ( j ) in tumor nodules. Statistical comparisons were performed using two-way ( a, i ) or one-way ( b, c, d, e, h, j ) analysis of variance (ANOVA) with Tukey’s multiple comparisons (liver enzyme measurement was compared to healthy controls). In all panels, each point corresponds to a single animal, with values shown as mean ± standard deviation.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a-j , B6C3F1 mice inoculated with 10 6 HM-1 tumor cells on day 0 were treated on days 10 with 20 µg of IL-12 as a free cytokine or conjugated to Mal NPs (UL and LbL). Two days after dosing ascites ( n = 6 animals/group) and i.p. tumor nodules (primarly omentum tissue, n = 4 animals/group) were harvested and processed for flow cytometry analysis. Shown are total counts of M1-like and M2-like macrophages (a) , logarithmic of M1-like to M2-like macrophages ratio ( b ), and total counts of PMN-MDSC ( c ), M-MDSC ( d) , and NK cells ( e ) in ascites fluid. Also shown are total counts of PMN-MDSC ( f ) and M-MDSC ( g ), logarithmic of M1-like to M2-like macrophages ratio ( h ), and total counts of M1-like and M2-like macrophages ( i ), and NK cells ( j ) in tumor nodules. Statistical comparisons were performed using two-way ( a, i ) or one-way ( b, c, d, e, h, j ) analysis of variance (ANOVA) with Tukey’s multiple comparisons (liver enzyme measurement was compared to healthy controls). In all panels, each point corresponds to a single animal, with values shown as mean ± standard deviation.

Techniques: Flow Cytometry, Standard Deviation

a , Schematic of the proposed mechanism of tumour-targeted IL-12 lipid conjugate dissemination from Mal LbL NPs. b , c , B6C3F1 mice inoculated with 10 6 HM-1-luc tumour cells on day 0 were administered fluorescently tagged Mal LbL of SAT LbL NPs carrying 20 µg of IL-12 on day 14. One day after dosing, the animals were euthanized, and the omentum containing tumour nodules was frozen in an OCT compound and then frozen, sectioned and stained for confocal microscopy analysis. Representative high-magnification confocal images of omental tumour nodules from Mal LbL ( b ) and SAT LbL ( c ). d – f , B6C3F1 mice (two cohorts for n = 7 and 5 animals per group per cohort) inoculated with 10 6 HM-1-luc tumour cells on day 0 were treated on day 7 with NP vehicle control (unloaded LbL), 20 µg of IL-12 as a free cytokine or conjugated to Mal LbL or SAT LbL. Experimental timeline ( d ), one representative in vivo IVIS whole-animal i.p. BLI readings from two independent experiments (mean ± s.d., n = 7 animals per group; e ) and overall survival ( f ). Statistics derived using all n from experiment with each animal as a data point. Statistical comparisons between survival curves were performed using a log-rank (Mantel–Cox) test.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a , Schematic of the proposed mechanism of tumour-targeted IL-12 lipid conjugate dissemination from Mal LbL NPs. b , c , B6C3F1 mice inoculated with 10 6 HM-1-luc tumour cells on day 0 were administered fluorescently tagged Mal LbL of SAT LbL NPs carrying 20 µg of IL-12 on day 14. One day after dosing, the animals were euthanized, and the omentum containing tumour nodules was frozen in an OCT compound and then frozen, sectioned and stained for confocal microscopy analysis. Representative high-magnification confocal images of omental tumour nodules from Mal LbL ( b ) and SAT LbL ( c ). d – f , B6C3F1 mice (two cohorts for n = 7 and 5 animals per group per cohort) inoculated with 10 6 HM-1-luc tumour cells on day 0 were treated on day 7 with NP vehicle control (unloaded LbL), 20 µg of IL-12 as a free cytokine or conjugated to Mal LbL or SAT LbL. Experimental timeline ( d ), one representative in vivo IVIS whole-animal i.p. BLI readings from two independent experiments (mean ± s.d., n = 7 animals per group; e ) and overall survival ( f ). Statistics derived using all n from experiment with each animal as a data point. Statistical comparisons between survival curves were performed using a log-rank (Mantel–Cox) test.

Techniques: Staining, Confocal Microscopy, Control, In Vivo, Derivative Assay

a , Illustration of liposome bilayer composition effect on lipid exchange rate with serum proteins. b , Intensity-weighted hydrodynamic size (Z-avg), number average size (#-avg), and PDI of NPs during synthesis as measured via DLS ( n = 3 technical replicates from representative batch, mean ± s.d.). c , Zeta potential of NPs during synthesis as measured via electrophoretic mobility in deionized water ( n = 3 technical replicates from representative batch, mean ± s.d.). d , Association of NP fluorescence with HM1 cells in vitro relative to unlayered NPs after 4 and 24 hours of incubation ( n = 6 replicates from two independent batches, mean ± s.d.). e , Representative confocal microscopy images of HM-1 cells dosed with SAT-LbL for 4 hours. f , Calculated IL-12 EC 50 of IL-12 compared to SAT-LbL NPs from HEK-Blue IL-12 assay (mean ± s.e.m.). g , Assessment of de-quenching from fluorophore detachment from unsat and SAT NPs when incubated with 100% FBS at 37 °C – curves represent the best fit of a two-phase decay model. h , Quantification of IL-12 retention with Mal-LbL or SAT-LbL upon incubation with 100% FBS ( n = 4-9 independent release experiments from two batches, mean ± s.d.) – curves represent the best fit of a two-phase decay model. Statistical comparisons performed in d using two-way analysis of variance (ANOVA) with Tukey’s multiple-comparisons. Data are representative of at least two independent experiments. Panel a created with BioRender.com .

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a , Illustration of liposome bilayer composition effect on lipid exchange rate with serum proteins. b , Intensity-weighted hydrodynamic size (Z-avg), number average size (#-avg), and PDI of NPs during synthesis as measured via DLS ( n = 3 technical replicates from representative batch, mean ± s.d.). c , Zeta potential of NPs during synthesis as measured via electrophoretic mobility in deionized water ( n = 3 technical replicates from representative batch, mean ± s.d.). d , Association of NP fluorescence with HM1 cells in vitro relative to unlayered NPs after 4 and 24 hours of incubation ( n = 6 replicates from two independent batches, mean ± s.d.). e , Representative confocal microscopy images of HM-1 cells dosed with SAT-LbL for 4 hours. f , Calculated IL-12 EC 50 of IL-12 compared to SAT-LbL NPs from HEK-Blue IL-12 assay (mean ± s.e.m.). g , Assessment of de-quenching from fluorophore detachment from unsat and SAT NPs when incubated with 100% FBS at 37 °C – curves represent the best fit of a two-phase decay model. h , Quantification of IL-12 retention with Mal-LbL or SAT-LbL upon incubation with 100% FBS ( n = 4-9 independent release experiments from two batches, mean ± s.d.) – curves represent the best fit of a two-phase decay model. Statistical comparisons performed in d using two-way analysis of variance (ANOVA) with Tukey’s multiple-comparisons. Data are representative of at least two independent experiments. Panel a created with BioRender.com .

Techniques: Zeta Potential Analyzer, Fluorescence, In Vitro, Incubation, Confocal Microscopy

a-b , B6C3F1 mice were inoculated with 10 6 HM-1-luc tumor cells on day 0 were administered fluorescently-tagged Mal-LbL of SAT-LbL NPs carrying 20 µg IL-12 on day 14 ( n = 4 animals/group). One day after dosing, animals were sacrificed, and the omentum containing tumor nodules was frozen in optimal cutting temperature (OCT) compound then frozen sectioned and stained for confocal microscopy analysis. Shown are representative confocal images of omental tumor nodules from Mal-LbL ( a ) and SAT-LbL ( b ) treated animals.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a-b , B6C3F1 mice were inoculated with 10 6 HM-1-luc tumor cells on day 0 were administered fluorescently-tagged Mal-LbL of SAT-LbL NPs carrying 20 µg IL-12 on day 14 ( n = 4 animals/group). One day after dosing, animals were sacrificed, and the omentum containing tumor nodules was frozen in optimal cutting temperature (OCT) compound then frozen sectioned and stained for confocal microscopy analysis. Shown are representative confocal images of omental tumor nodules from Mal-LbL ( a ) and SAT-LbL ( b ) treated animals.

Techniques: Staining, Confocal Microscopy

a-d B6C3F1 mice ( n = 5 animals/group) inoculated with 10 6 HM-1 tumor cells on day 0 were treated on day 10 with 20 µg of IL-12 as a free cytokine or conjugated to Mal-LbL NPs. Two days after dosing ascites and i.p. tumor nodules (primarily omentum tissue) were harvested and processed for flow cytometry analysis. Shown are quantitation of the percentage of T regs among T cells in ascites ( a ), PD1 positive CD4 T cell ( b ) and CD8 T cell ( c ) in ascites, CD25 + FoxP3 − of CD4 T cells ( d ) in ascites, CD25 + of CD8 T cells ( e) in ascites, T regs of T cells in tumor ( f ), CD25 + FoxP3 − of CD4 T cells in tumor ( g ), PD1 + TIM3 − of CD4 T cells in tumors ( h ), PD1 + TIM3 + of CD4 T cells in tumors ( i ), PD1 + TIM3 − of CD8 T cells in tumors ( j ), PD1 + TIM3 + of CD8 T cells in tumors ( k ), and CD25 + of CD8 T cells ( l ). m , B6C3F1 mice ( n = 10 animals/group) inoculated with 10 6 HM-1-luc tumor cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal-LbL Mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 i.p. on days 8 and 15. Shown are the relative body weight of mice (black arrows indicate IL-12 treatment and blue indicate CPI). None of the treated mice showed significant drop in bodyweight compared to dextrose control. n-p , B6C3F1 mice (2 cohorts with n = 5 animals/group/cohort) inoculated with 10 6 HM-1-luc tumor cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal-LbL or Ni-LbL. Mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 i.p. on days 8 and 15. Shown are the in vivo IVIS whole-animal i.p. BLI readings ( mean ± s.d., n) . On day 150, surviving mice were rechallenged with either 3×10 5 ( o ) or 10 6 ( p ) HM-1-luc tumor cells i.p. Shown are the in vivo IVIS whole-animal i.p. BLI readings post rechallenge ( mean ± s.d). In all panels, each point corresponds to a single animal, with values shown as mean ± standard deviation.

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a-d B6C3F1 mice ( n = 5 animals/group) inoculated with 10 6 HM-1 tumor cells on day 0 were treated on day 10 with 20 µg of IL-12 as a free cytokine or conjugated to Mal-LbL NPs. Two days after dosing ascites and i.p. tumor nodules (primarily omentum tissue) were harvested and processed for flow cytometry analysis. Shown are quantitation of the percentage of T regs among T cells in ascites ( a ), PD1 positive CD4 T cell ( b ) and CD8 T cell ( c ) in ascites, CD25 + FoxP3 − of CD4 T cells ( d ) in ascites, CD25 + of CD8 T cells ( e) in ascites, T regs of T cells in tumor ( f ), CD25 + FoxP3 − of CD4 T cells in tumor ( g ), PD1 + TIM3 − of CD4 T cells in tumors ( h ), PD1 + TIM3 + of CD4 T cells in tumors ( i ), PD1 + TIM3 − of CD8 T cells in tumors ( j ), PD1 + TIM3 + of CD8 T cells in tumors ( k ), and CD25 + of CD8 T cells ( l ). m , B6C3F1 mice ( n = 10 animals/group) inoculated with 10 6 HM-1-luc tumor cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal-LbL Mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 i.p. on days 8 and 15. Shown are the relative body weight of mice (black arrows indicate IL-12 treatment and blue indicate CPI). None of the treated mice showed significant drop in bodyweight compared to dextrose control. n-p , B6C3F1 mice (2 cohorts with n = 5 animals/group/cohort) inoculated with 10 6 HM-1-luc tumor cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal-LbL or Ni-LbL. Mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 i.p. on days 8 and 15. Shown are the in vivo IVIS whole-animal i.p. BLI readings ( mean ± s.d., n) . On day 150, surviving mice were rechallenged with either 3×10 5 ( o ) or 10 6 ( p ) HM-1-luc tumor cells i.p. Shown are the in vivo IVIS whole-animal i.p. BLI readings post rechallenge ( mean ± s.d). In all panels, each point corresponds to a single animal, with values shown as mean ± standard deviation.

Techniques: Flow Cytometry, Quantitation Assay, Control, In Vivo, Standard Deviation

a – d , B6C3F1 mice ( n = 5 animals per group) inoculated with 10 6 HM-1 tumour cells on day 0 were treated on day 10 with 20 µg of IL-12 as a free cytokine or conjugated to Mal LbL NPs. Two days after dosing, ascites and i.p. tumour nodules (primarily omentum tissue) were harvested and processed for flow cytometry analysis. Flow plots of PD1 and TIM3 expression on T cells (CD45 + CD3 + ) in tumour nodules ( a ), quantitation of PD1 MFI of T cells in tumour nodules ( b ), representative fluorescence histogram plots of CD25 expression on CD8 T cells (CD45 + CD3 + CD8 + ) in tumour nodules ( c ) and quantitation of CD25 MFI of CD8 T cells in tumour nodules ( d ). e , f , B6C3F1 mice (two cohorts with n = 5 animals per group per cohort) inoculated with 10 6 HM-1-luc tumour cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal LbL or Ni LbL. Mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 i.p. on days 8 and 15. Experimental timeline ( e ) and overall survival ( f ). On day 150, the surviving mice were rechallenged with either 3 × 10 5 ( g ) or 10 6 ( h ) i.p. HM-1-luc tumour cells. Percentage of mice per group that survived the rechallenge is shown. i , B6C3F1 mice (one cohort with n = 10 animals per group) inoculated with 10 6 HM-1-luc tumour cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine, Mal LbL, SAT LbL or Mal LbL in combination with depletion antibodies. With an exception of the dextrose group, mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 via i.p. administration on days 8 and 15. Overall survival curves are shown. j , k , C57BL/6 mice (one cohort of n = 10 animals per group) were inoculated via i.p. administration with 5 × 10 6 ID8 ( Trp53 − / − ; Brca2 − / − ; j ) or 10 6 KPCA tumour cells ( k ) on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal LbL. Mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 via i.p. administration on days 8 and 15. Statistics derived using all n from experiment with each animal as a data point. Overall survival curves are shown. P values were determined by one-way ANOVA followed by Tukey’s multiple comparison test ( b and d ) and the log-rank (Mantel–Cox) test ( f and i – k ).

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a – d , B6C3F1 mice ( n = 5 animals per group) inoculated with 10 6 HM-1 tumour cells on day 0 were treated on day 10 with 20 µg of IL-12 as a free cytokine or conjugated to Mal LbL NPs. Two days after dosing, ascites and i.p. tumour nodules (primarily omentum tissue) were harvested and processed for flow cytometry analysis. Flow plots of PD1 and TIM3 expression on T cells (CD45 + CD3 + ) in tumour nodules ( a ), quantitation of PD1 MFI of T cells in tumour nodules ( b ), representative fluorescence histogram plots of CD25 expression on CD8 T cells (CD45 + CD3 + CD8 + ) in tumour nodules ( c ) and quantitation of CD25 MFI of CD8 T cells in tumour nodules ( d ). e , f , B6C3F1 mice (two cohorts with n = 5 animals per group per cohort) inoculated with 10 6 HM-1-luc tumour cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal LbL or Ni LbL. Mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 i.p. on days 8 and 15. Experimental timeline ( e ) and overall survival ( f ). On day 150, the surviving mice were rechallenged with either 3 × 10 5 ( g ) or 10 6 ( h ) i.p. HM-1-luc tumour cells. Percentage of mice per group that survived the rechallenge is shown. i , B6C3F1 mice (one cohort with n = 10 animals per group) inoculated with 10 6 HM-1-luc tumour cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine, Mal LbL, SAT LbL or Mal LbL in combination with depletion antibodies. With an exception of the dextrose group, mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 via i.p. administration on days 8 and 15. Overall survival curves are shown. j , k , C57BL/6 mice (one cohort of n = 10 animals per group) were inoculated via i.p. administration with 5 × 10 6 ID8 ( Trp53 − / − ; Brca2 − / − ; j ) or 10 6 KPCA tumour cells ( k ) on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal LbL. Mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 via i.p. administration on days 8 and 15. Statistics derived using all n from experiment with each animal as a data point. Overall survival curves are shown. P values were determined by one-way ANOVA followed by Tukey’s multiple comparison test ( b and d ) and the log-rank (Mantel–Cox) test ( f and i – k ).

Techniques: Flow Cytometry, Expressing, Quantitation Assay, Fluorescence, Derivative Assay, Comparison

a-d , B6C3F1 mice inoculated with 10 6 HM-1-luc tumor cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal-LbL Mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 i.p. on days 8 and 15. Mice were euthanized before tumor inoculation (healthy n = 5), prior to the first dose (day 7, n = 3 animals), on day 65 ( n = 5 animals), or on day 130 ( n = 4 animals). Organs were harvested and processed for histopathologic evaluation. Shown are representative images of the primary finding in the omentum/pancreasin naive mice ( a ), day 7 mice showing tumor, inflammation and pancreatic degeneration ( b ), day 65 mice showing inflammation and pancreatic degeneration ( c ), and day 130 mice showing recovered omentum/pancreas ( d ).

Journal: Nature Materials

Article Title: IL-12-releasing nanoparticles for effective immunotherapy of metastatic ovarian cancer

doi: 10.1038/s41563-025-02390-9

Figure Lengend Snippet: a-d , B6C3F1 mice inoculated with 10 6 HM-1-luc tumor cells on day 0 were treated on days 7 and 14 with 20 µg of IL-12 as a free cytokine or conjugated to Mal-LbL Mice were also treated with 250 µg of anti-PD1 and 100 µg of anti-CTLA4 i.p. on days 8 and 15. Mice were euthanized before tumor inoculation (healthy n = 5), prior to the first dose (day 7, n = 3 animals), on day 65 ( n = 5 animals), or on day 130 ( n = 4 animals). Organs were harvested and processed for histopathologic evaluation. Shown are representative images of the primary finding in the omentum/pancreasin naive mice ( a ), day 7 mice showing tumor, inflammation and pancreatic degeneration ( b ), day 65 mice showing inflammation and pancreatic degeneration ( c ), and day 130 mice showing recovered omentum/pancreas ( d ).

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