A5115598785- Immune Cell Function and Interaction
- Drug Transport and Resistance Mechanisms
- Neuroblastoma Research and Treatments
- Neuroendocrine Tumor Research Advances
- Cancer, Hypoxia, and Metabolism
- Glioma Diagnosis and Treatment
- Immune cells in cancer
- Adrenal and Paraganglionic Tumors
- Cancer therapeutics and mechanisms
- Pancreatic and Hepatic Oncology Research
- Immune Response and Inflammation
- Pediatric Hepatobiliary Diseases and Treatments
- T-cell and B-cell Immunology
- Hepatocellular Carcinoma Treatment and Prognosis
- Immunotherapy and Immune Responses
- Sarcoma Diagnosis and Treatment
- Hepatitis B Virus Studies
- Gut microbiota and health
- Plant Stress Responses and Tolerance
- Neurofibromatosis and Schwannoma Cases
- Ocular Oncology and Treatments
- Chemical Reactions and Isotopes
- Allelopathy and phytotoxic interactions
- Tuberous Sclerosis Complex Research
- Diet and metabolism studies
Beijing Children’s Hospital
2009-2024
Capital Medical University
2009-2024
Wuxi People's Hospital
2024
Nanjing Medical University
2024
China Agricultural University
2024
Beijing Friendship Hospital
2024
Capital University
2019-2020
<p>Supplementary Figure 2 Gating strategies of FACS analysis and the expression pattern TIGIT in NB. (A) for circulating γδT cells. (B) tumor infiltrating immune (C) (D) The CD96 CD112R peripheral blood (PB) cells from healthy controls (HC, n = 10) neuroblastoma patients (NB, 18). proportions are shown left panel statistically representative histograms right with mean fluorescence intensity (MFI) labeled corresponding histograms. (E) Using GSE49711 GEO database R2: Genomics Analysis...
<p>Supplementary Figure 3 The expression analysis of CD112 and CD155 in NB. (A) (B) NB cell lines primary tumor tissues are identified using qPCR Western blot. (C) (D) FACS. (E) Soluble serum is detected by ELISA (HC, n = 17, NB, 22) further analyzed between different groups divided according to INSS stage, risk MYCN status. (F) (G) (H) (I) (J) 21, 35) results expressed as the means ± SEMs from at least three independent experiments. Significant differences represented ns no...
<p>Supplementary Figure 13 The effects of CD155 on γδT-cell-cytotoxicity against neuroblastoma, and proliferation migration NB cells. (A) (B) α-TIGIT is used in the rhCD155 treatment γδT DNAM-1 detected by FACS Western blot. (C-F) to treat cells for 24 h. TIGIT, CD96 CD112R are (left) or blot MFI labeled representative histograms (right). (G) γδT-cell numbers counted statistically analyzed during vitro culture presence rhCD155. (H) CFSE assays performed (right) (left). histograms. (I)...
<p>Supplementary Figure 9 The characteristics of NB tumors and immune cells in CD112/CD155 high-ratio low-ratio groups revealed by bulk RNA sequencing analysis. (A) A total 3537 DEGs between (grouping according to the ratio median violin plot) are identified analysis with 38 tumor samples illustrated volcano plot. (B) expression genes related neuroblastoma KEGG pathway transcriptional misregulation cancer is heatmap. (C) (D) enriched top 20 pathways or shown dot plots. (E) (F) 30 GO...
<p>Supplementary Figure 14 The effects of soluble CD155 or on NB cells DNAM-1, PD-1, TIM-3 and TIGIT expression levels in γδT during the interaction with microenvironment. (A-C) α-CD155 is used co-culture SH-SY5Y cells. are detected by FACS after 24 h (left) MFI labeled representative histograms (right). (D-G) primary tumor (H-K) TTCS treatment results expressed as means ± SEMs from at least three independent experiments. Significant differences between groups represented ns no...
<p>Supplementary Figure 4 Quality control and annotation genes for NB tumor spatial transcriptome analysis. (A) Violin plots heatmaps showing the number of features, RNA counts percent mitochondrial transcripts found in sample following quality control. (B) UMAP plot major cell types Dots represent individual cells, colors different populations. (C) dot expression annotating identified tumor.</p>
<p>Supplementary Figure 5 The expression of CD112 and CD155 in tumor para-tumor tissue sections detected by immunohistochemistry.</p>
<p>Supplementary Figure 6 The cell-cycle scores and expression of functional genes in tumor cells γδT spatial transcriptome analysis scRNA sequencing analysis. (A) Cell-cycle score (B) Spatial distribution proliferating markers. (C) (D) Violin plots showing the levels neuroblastoma related (E) (F) GSVA (G) DNAM-1, TIGIT, CD96, PD-1, TIM-3, IFN-γ, granzyme B perforin analysis.</p>
<p>Supplementary Figure 10 Identification of Tet-on system, ROC curves and survival different groups for NB tumor. (A) (B) The variation CD112 CD155 expression in SK-N-BE2 cells treated with concentration doxycycline (0, 2 nM, 20 200 μM, μM) is detected by qPCR Western blot. (C) are generated CD112, CD155, CD112/CD155 ratio to predict patient death, tumor risk or MYCN amplification. results expressed as the means ± SEMs from at least three independent experiments. Significant...
<p>Supplementary Figure 11 Low-dose doxorubicin affects the expression of CD112 and CD155 in NB cells. (A) The apoptosis SH-SY5Y SK-N-BE2 cells treated with low-dose (0.1 μM) is detected by FACS (left) AV+ proportion labeled representative histograms. (B-F) or primary tumor are Western blot. MFI (G) (H) cytotoxicity assays performed to γδT-cell against pretreated 0.1 μM doxorubicin. results expressed as means ± SEMs from at least three independent experiments. Significant differences...
<p>Supplementary Figure 7 Metabolic states of high-ratio and low-ratio tumor cells. (A) Enriched metabolic reactions in cells revealed by COMPASS analysis. (B) Changes KEGG pathways spatial metabolome (C) The heatmaps representing metabolites.</p>
<p>Supplementary Figure 12 The effects of CD112 on γδT-cell-cytotoxicity against neuroblastoma, and proliferation migration NB cells. (A-D) rhCD112 is used to treat γδT cells for 24 h. TIGIT, CD96 CD112R are detected by FACS (left) or Western blot MFI labeled in the representative histograms (right). (E) γδT-cell numbers counted statistically analyzed during vitro culture presence rhCD112. (F) CFSE assays performed (right) (left). histograms. (G) pre-treated with h subjected calcium...
<p>Supplementary Figure 8 Enriched pathways and functional characteristics of CD8 T NK cells. (A) The enriched KEGG in cells from high-ratio low-ratio groups are shown dot plots. size enrichment analysis represents gene numbers. (B) (C) Cell-cycle score GSVA (D) Violin plots showing the expression levels DNAM-1, TIGIT, CD96, PD-1, TIM-3, IFN-γ, granzyme B perforin (E) (F) (G) (H) cells.</p>
<p>Supplementary Figure 1 Identified major cell types, proportion analysis and CD112 CD155 expression in NB tumor scRNA-seq analysis. (A) Violin plots showing the number of features, RNA counts percent mitochondrial transcripts following quality control. (B) average genes for annotating types identified tumors. (C) UMAP distribution each sample. (D) Bar proportions (E) levels CD155.</p>
<div>Abstract<p>The dynamic interplay between tumor cells and γδT within the microenvironment significantly influences disease progression immunotherapy outcome. In this study, we delved into modulation of γδT-cell activation by cell ligands CD112 CD155, which interact with activating receptor DNAM-1 on cells. Spatial single-cell RNA sequencing, as well spatial metabolomic analysis, from neuroblastoma revealed that expression levels localization CD155 varied across tumors,...
<p>Supplementary Figure 3 The expression analysis of CD112 and CD155 in NB. (A) (B) NB cell lines primary tumor tissues are identified using qPCR Western blot. (C) (D) FACS. (E) Soluble serum is detected by ELISA (HC, n = 17, NB, 22) further analyzed between different groups divided according to INSS stage, risk MYCN status. (F) (G) (H) (I) (J) 21, 35) results expressed as the means ± SEMs from at least three independent experiments. Significant differences represented ns no...
<p>Supplementary Figure 2 Gating strategies of FACS analysis and the expression pattern TIGIT in NB. (A) for circulating γδT cells. (B) tumor infiltrating immune (C) (D) The CD96 CD112R peripheral blood (PB) cells from healthy controls (HC, n = 10) neuroblastoma patients (NB, 18). proportions are shown left panel statistically representative histograms right with mean fluorescence intensity (MFI) labeled corresponding histograms. (E) Using GSE49711 GEO database R2: Genomics Analysis...
<p>Supplementary Figure 5 The expression of CD112 and CD155 in tumor para-tumor tissue sections detected by immunohistochemistry.</p>
<p>Supplementary Figure 6 The cell-cycle scores and expression of functional genes in tumor cells γδT spatial transcriptome analysis scRNA sequencing analysis. (A) Cell-cycle score (B) Spatial distribution proliferating markers. (C) (D) Violin plots showing the levels neuroblastoma related (E) (F) GSVA (G) DNAM-1, TIGIT, CD96, PD-1, TIM-3, IFN-γ, granzyme B perforin analysis.</p>
<p>Supplementary Figure 4 Quality control and annotation genes for NB tumor spatial transcriptome analysis. (A) Violin plots heatmaps showing the number of features, RNA counts percent mitochondrial transcripts found in sample following quality control. (B) UMAP plot major cell types Dots represent individual cells, colors different populations. (C) dot expression annotating identified tumor.</p>
<p>Supplementary Figure 10 Identification of Tet-on system, ROC curves and survival different groups for NB tumor. (A) (B) The variation CD112 CD155 expression in SK-N-BE2 cells treated with concentration doxycycline (0, 2 nM, 20 200 μM, μM) is detected by qPCR Western blot. (C) are generated CD112, CD155, CD112/CD155 ratio to predict patient death, tumor risk or MYCN amplification. results expressed as the means ± SEMs from at least three independent experiments. Significant...
<p>Supplementary Figure 14 The effects of soluble CD155 or on NB cells DNAM-1, PD-1, TIM-3 and TIGIT expression levels in γδT during the interaction with microenvironment. (A-C) α-CD155 is used co-culture SH-SY5Y cells. are detected by FACS after 24 h (left) MFI labeled representative histograms (right). (D-G) primary tumor (H-K) TTCS treatment results expressed as means ± SEMs from at least three independent experiments. Significant differences between groups represented ns no...
<p>Supplementary Figure 12 The effects of CD112 on γδT-cell-cytotoxicity against neuroblastoma, and proliferation migration NB cells. (A-D) rhCD112 is used to treat γδT cells for 24 h. TIGIT, CD96 CD112R are detected by FACS (left) or Western blot MFI labeled in the representative histograms (right). (E) γδT-cell numbers counted statistically analyzed during vitro culture presence rhCD112. (F) CFSE assays performed (right) (left). histograms. (G) pre-treated with h subjected calcium...
<p>Supplementary Figure 11 Low-dose doxorubicin affects the expression of CD112 and CD155 in NB cells. (A) The apoptosis SH-SY5Y SK-N-BE2 cells treated with low-dose (0.1 μM) is detected by FACS (left) AV+ proportion labeled representative histograms. (B-F) or primary tumor are Western blot. MFI (G) (H) cytotoxicity assays performed to γδT-cell against pretreated 0.1 μM doxorubicin. results expressed as means ± SEMs from at least three independent experiments. Significant differences...