Journal: Nature Communications

Article Title: Hypoxia induces HIF1α-dependent epigenetic vulnerability in triple negative breast cancer to confer immune effector dysfunction and resistance to anti-PD-1 immunotherapy

doi: 10.1038/s41467-022-31764-9

Figure Lengend Snippet: a Schematic diagram showing the establishment of humanized mice (humice) with human immune system reconstituted in NIKO mice. The presence of human CD45 + cells, NK cells, CD4 + and CD8 + T cells in the mice’s peripheral system was validated by flow cytometry. b Primary LM2 tumor size in humice (control, n = 14; Keytruda, n = 14; ENT, n = 12; PX478, n = 14; ENT + Keytruda, n = 16; PX478 + Keytruda, n = 16) and NIKO mice (control, n = 10; ENT + Keytruda, n = 10; PX478 + Keytruda, n = 10), at Day 21 of treatments. c Lung metastasis of humice (control, n = 6; Keytruda, n = 6; ENT, n = 6; PX478, n = 6; ENT + Keytruda, n = 7; PX478 + Keytruda, n = 7) and NIKO mice (control, n = 5; ENT + Keytruda, n = 5; PX478 + Keytruda, n = 5) bearing LM2 tumors at Day 35 assessed by bioluminescence (BLI) measurement. d Representative bioluminescence (BLI) images showing the lung metastasis of humice and NIKO mice. e Flow cytometric analysis of LM2 tumors harvested from humanized mice. IFNγ, TNFα, and granzyme B expression was examined in tumor-infiltrating human CD8 + T cells and NK cells. N = 5 for each group. f Flow cytometry analysis of LM2 tumors harvested from humanized mice. Expressions of human PD-L1 and PD-L2 were examined in total living cells dissociated from LM2 tumors. N = 5 for each group. Quantification data of flow cytometry ( e , f ) are presented as a box and whiskers, with median values and whiskers of minimum and maximum values. Data for b and c were presented as mean ± SD . P values were determined by one-way ( e , f ) or two-way ( b , c ) ANOVA with Turkey’s test. Source data are provided as a source data file.

Article Snippet: The following antibodies were used for staining, anti-activated pimonidazole FITC antibody (Hypoxyprobe, CAT# HP2-200kit, dilution 1:200), anti-mouse HIF1α APC antibody (R&D Systems, CAT# IC1935A, dilution 1:50), anti-mouse CD3 BV421 antibody (BD Biosciences, CAT# 564008, dilution 1:100), anti-mouse CD45 Percp-Vio700 antibody (Miltenyi Biotec, CAT# 130-110-663, dilution 1:100) anti-mouse CD8 APC-Vio770 antibody (Miltenyi Biotec, CAT# 130-120-737, dilution 1:100), anti-mouse Nkp46 APC antibody (Miltenyi Biotec, CAT# 130-112-202, dilution 1:100), anti-mouse CD4 BV650 antibody (Biolegend, CAT# 563747, dilution 1:100), anti-mouse TIM-3 BV711 antibody (Biolegend, CAT# 119727, dilution 1:100), anti-mouse PD-1 PE-Vio770 (Miltenyi Biotec, CAT# 130-120-391, dilution 1:100), anti-mouse IFNγ PE (Miltenyi Biotec, CAT# 130-117-352, dilution 1:100), anti-mouse TNFα BV711 (BD Biosciences, CAT# 563944, dilution 1:100), anti-mouse/human granzyme B FITC (Miltenyi Biotec, Cat#130-118-430, dilution 1:100), anti-mouse PD-L1 BV786 antibody (BD Biosciences, CAT# 741014, dilution 1:100), anti-mouse PD-L2 FITC antibody (Miltenyi Biotec, Cat# 130-102-222, dilution 1:100), anti-human CD45 FITC antibody (BD Biosciences, CAT# 304006, dilution 1:100), anti-human CD3 PE antibody (Biolegend, CAT# 300308, dilution 1:100) anti-human CD8 APC-Cy7 antibody (BD Biosciences, CAT# 557834, dilution 1:100), anti-human CD56 BV711 antibody (Biolegend, CAT# 318336, dilution 1:100), anti-human CD4 APC antibody (Biolegend, CAT# 300514, dilution 1:100), anti-human IFNγ BV785 (Biolegend, CAT# 502542, dilution 1:100), anti-human TNFα BV650 (Biolegend, CAT# 502398, dilution 1:100), anti-human Granzyme B BV421 (BD Biosciences, Cat# 563389, dilution 1:100), anti-human PD-L1 PE-Cy7 antibody (Biolegend, CAT# 374506, dilution 1:100), anti-human PD-L2 PE antibody (Miltenyi Biotec, CAT# 130-098-530, dilution 1:100).

Techniques: Flow Cytometry, Control, Expressing

Journal: bioRxiv

Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling

doi: 10.64898/2026.03.24.712780

Figure Lengend Snippet: ( A ) Mice were inoculated with 3×10 6 BPPNM cells IP and dosed with 20 μg of STING or Scr conjugate delivered by LNP IP at 10, 13, 16, and 19 days after inoculation. Omental tumor was collected on day 20 for analysis by flow cytometry. Groups included N = 5 mice. ( B ) Cell populations in tumor after treatment, displaying percentage of CD45 + cells made up by T cells (CD3 + ), CD8 + T cells, CD4 + T cells, NK cells (CD3 - NK1.1 + ), B cells (CD19 + ), Macrophages (F4/80 + ), DCs (CD11c + MHCII + ), and MDSCs (CD11b + Gr-1 hi ). Percentage of CD45 - cells made up by BPPNM cancer cells (GFP + ) are also displayed. ( C-D ) Polarization of Macrophages (F4/80 + ), showing MFI and representative distributions of ( C ) CD86 and ( D ) CD206. ( E ) Activation of DCs (CD11c + MHCII + ), showing MFI and representative distributions of CD86. ( F ) Expression of PD-1 on CD8 + T cells, showing percentage of cells that are PD-1 + and representative distributions of PD-1. P values computed with a one-way ANOVA followed by Tukey’s post-hoc test are displayed above each figure. All flow data is represented as mean ± SD. ( G ) Mice were inoculated with 3×10 6 BPPNM cells IP and dosed with 20 μg of STING or Scr conjugate delivered by LNP IP at 10, 13, 16, 19, and 22 days after inoculation. A subset of groups were additionally treated with 100 μg αPD-1 antibody at 11 and 17 days after inoculation. Groups included N = 6 (PBS, STING LNP, Scr LNP), N = 5 (STING LNP + αPD-1), or N = 4 (PBS + αPD-1, Scr LNP + αPD-1) mice. ( H ) Survival plot with P values determined by log(rank) (Mantel–Cox) test. ( I-K ) Tumor burden measured by IVIS, displaying ( I ) geometric mean ± SD bioluminescent intensity over the treatment period, ( J ) an image of the mouse with the median bioluminescent intensity from each group at the measurement following the end of treatment (day 24), and ( K ) individual mouse bioluminescent intensity values.

Article Snippet: In groups specified in figure captions, mice were also administered a 100 μg dose of anti-mouse PD-1 antibody (Bio X Cell #BE0273 InVivoMAb, Clone 29F.1A12) in 200 μL of 1× PBS on days 11 and 17 after tumor inoculation.

Techniques: Flow Cytometry, Activation Assay, Expressing

Journal: Microbiome

Article Title: Tumor-colonized Streptococcus mutans metabolically reprograms tumor microenvironment and promotes oral squamous cell carcinoma.

doi: 10.1186/s40168-024-01907-9

Figure Lengend Snippet: Fig. 5 Single-cell transcriptomic landscape reveals the shift of the cellular types during oral carcinogenesis, and KYNA is a predominant driving force for expansions and infiltrations of neutrophils in the TME of OSCC. A Overview of the workflow for scRNA-seq analyses of OLK and OSCC rat tongue tissues. B UMAP plot of the clustering results for 13 major cell types from OLK and OSCC tissues. C Stacked histogram of the percentages of different cells from OLK and OSCC tissues. D Quantitative analysis of neutrophils, macrophages, and T cells in non-epithelial cells in OLK and OSCC tissues. E H&E and IHC staining for CD11b in OLK and OSCC tissues of experimental rats. Scale bar: 600 µm for 4 × magnification and 300 µm for 10 × magnification. F Representative IHC staining images and statistical analysis of CD16 in saline and KYNA-treated 4NQO rats. Scale bars: 200 µm for 10 × magnification and 50 µm for 40 × magnification. G Representative flow cytometry dot plots and statistical analysis of CD11b + CD16 + neutrophils from the peripheral blood of OSCC patients after KYNA treatment. H Relative SLC7A8 expression in neutrophils from OSCC patients after KYNA treatment detected by RT-qPCR. I Representative flow cytometry dot plots and statistical analysis of AHR + neutrophils from OSCC patients after treatments with KYNA in indicated concentrations. In D, unpaired Student’s t-test; E and F, Mann– Whitney U test; G, H, and I, one-way ANOVA

Article Snippet: At week 20, the rats were sacrificed, and the tongues were dissected, and a longitudinal mid-lingual incision was made. (4) IL-1β/PD-L1monoclonal antibody (mAb) intervention (8 rats/group): (A) 4NQO + IL-1β mAb (200 μg/rat, BE0246, cloneB122, Bioxcell, USA); (B) 4NQO + KYNA + IL-1β mAb; (C) 4NQO + PD-L1 mAb (200 μg/rat, BE0383, clone 368A.4H1, Bioxcell, USA); (D) 4NQO + KYNA + PD-L1 mAb; (E) 4NQO + control IgG (200 μg/rat, BE0091, Bioxcell, USA).

Techniques: Immunohistochemistry, Saline, Flow Cytometry, Expressing, Quantitative RT-PCR, MANN-WHITNEY