Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Distinct immune signatures for predicting the immunotherapy efficacy of esophageal squamous cell carcinoma or adenocarcinoma

doi: 10.1007/s00262-024-03904-1

Figure Lengend Snippet: Detecting unique cytokines linked to clinical outcomes in esophageal squamous cell carcinoma and adenocarcinoma separately. a Venn diagram shows genes in the cytokine-cytokine receptor pathway for ESCC and EAC from TCGA, annotated with gene counts in distinct and overlapping categories. b Overall survival of patients with TCGA-ESCC and different expression levels of TNFSF10 and CXCL14. c TNFSF10 and CXCL14 expression comparison between early and advanced ESCC tissues (TCGA). d Survival probabilities of patients with EAC classified by CXCL8 and IL17RB expression levels (TCGA). e CXCL8 and IL17RB expression during EAC progression from early to advanced stages. f TNFSF10 and CXCL14 expression in early and advanced ESCC tissues of Chinese patients. g, h The left panel presents immunohistochemical images illustrating TNFSF10 or CXCL14 staining in tissue microarrays obtained from patients diagnosed with ESCC, arranged sequentially from left to right based on ascending expression levels quantified by the H-score. On the far left, the lowest expression level is shown, followed by a sample in the lowest 33.33 percentile, then a moderate expression sample in the next 33.33 percentile, and finally, the highest expression level on the far right. The right panel displays survival curves stratified by the expression of TNFSF10 and CXCL14, categorized into low and high expression groups based on their H-scores. i, j The left panel displays immunohistochemical staining for CXCL8 or IL17RB in tissue microarrays from patients with EAC, with samples ordered from left to right by increasing H-scores. The sequence starts with the lowest expression level, followed by a sample in the first 33.33 percentile, then a sample with moderate expression in the next percentile, and ends with the highest expression level on the far right. P values were calculated using the Mann–Whitney (log-rank) test. Early stage: Stage 0-II; Advanced stage: Stage III-IV; ns: no significance; * P < 0.05, Mann–Whitney test

Article Snippet: The tissue sections were incubated with primary antibodies against TNFSF10 (Catalog No. ab2056; Abcam), CXCL14 (catalog no. ab137541; Abcam), CXCL8 (catalog no. ab106350; Abcam), IL17RB (Catalog No. AF1207; R&D Systems), CXCL10 (Catalog No. ab306587; Abcam, Cambridge, UK), HIF1α (catalog no. ab51608; Abcam), CSF2 (catalog no. ab316862; Abcam), and NOS2 (catalog no. ab283655; Abcam) (diluted at 1:500 in phosphate-buffered saline) overnight at 4℃.

Techniques: Expressing, Comparison, Immunohistochemical staining, Staining, Sequencing, MANN-WHITNEY

Journal: Cancer Gene Therapy

Article Title: SEMA7A-mediated juxtacrine stimulation of IGFBP-3 upregulates IL-17RB at pancreatic cancer invasive front

doi: 10.1038/s41417-024-00849-6

Figure Lengend Snippet: Matrigel invasion assays of HPAF-II ( A ) and MIA PaCa-2 ( B ) cells from parental or IL-17RB Del clones. Each dot represents the datum from one independent experiment (n = 4). Values are expressed as mean ± SD. ***, P < 0.001 (two-tailed Student’s t-test). C Western blotting analysis of proteins harvested from parental or IL-17RB Del tumor cells (HPAF-II or MIA PaCa-2). D Western blotting analysis of proteins harvested from HPAF-II or MIA PaCa-2 cells transduced with lentiviral pLAS5w (vector) or pLAS5w-IL17RB-Flag (IL17RB-Flag). E Representative IHC images of IL-17RB (green signal), SNAI2 (red signal) and Hoechst 33342 (blue signal) expression in FFPE samples of PDAC patients. Scale bar, 100 μm. F Relative intensity of SNAI2 in IL-17RB low and high expressing regions. IL-17RB low expressing regions were defined as IL-17RB staining expressed in <10% tumor cells per high power field, and IL-17RB high expressing regions were defined as IL-17RB staining expressed in ≥50% tumor cells per high power field. Each dot represents the datum from one high-power fields (n = 8). Values are expressed as mean ± SD. **, P < 0.01 (two-tailed Student’s t-test). G Representative IHC images of IL-17RB (green signal), VIM (red signal), Hoechst 33342 (blue signal) expression in FFPE samples of PDAC patients. Scale bar, 100 μm. H Relative intensity of VIM in IL-17RB low and high expressing regions. IL-17RB low expressing regions were defined as IL-17RB staining expressed in <10% tumor cells per high power field, and IL-17RB high expressing regions were defined as IL-17RB staining expressed in ≥50% tumor cells per high power field. Each dot represents the datum from one high-power field n = 10). Values are expressed as mean ± SD. *, P < 0.05 (two-tailed Student’s t-test). I , J Western blotting analysis (I) and Matrigel invasion assays (J) of cells from parental, IL-17RB Del (none), IL-17RB Del transduced with pLAS5w (vector), and IL-17RB Del transduced with pLAS5w-SNAI2 (SNAI2). Each dot represents the datum from one independent experiment (n = 4). Values are expressed as mean ± SD. ****, P < 0.0001 (one-way ANOVA).

Article Snippet: The cDNA clone of human IL17RB in pCMV6-XL5 was obtained from OriGene. pLAS5w-IL17RB-Flag was constructed by insertion of a 1509-bp fragment of the full-length human IL17RB cDNA (NM_018725.4) and Flag at NheI site and Sbf1 site of pLAS5w.Pbsd-L-tRFP (National RNAi Core Facility, Taiwan) vector.

Techniques: Clone Assay, Two Tailed Test, Western Blot, Transduction, Plasmid Preparation, Expressing, Staining