Journal: Journal of Experimental & Clinical Cancer Research : CR
Article Title: S100A9 + CD14 + monocytes contribute to anti-PD-1 immunotherapy resistance in advanced hepatocellular carcinoma by attenuating T cell-mediated antitumor function
doi: 10.1186/s13046-024-02985-1
Figure Lengend Snippet: Endogenous S100A9 enhances PD-L1 expression in monocytes to inhibit T-cell cytotoxicity. a KEGG pathway enrichment analysis of DEGs between S100A9 + CD14 + monocytes and other circulating cells. b Percentage of S100A9 + cells (left) and PD-L1 + cells (right) in PBMC from healthy donors (HD, n = 5) and patients with hepatocellular carcinoma (HCC, n = 7), colorectal cancer (CRC, n = 5), and biliary tract cancer (BTC, n = 7). c-e PD-L1 levels in S100A9 high or S100A9 low cells in CD14 + monocytes from patients with HCC, CRC, and BTC. f-h Correlation analysis between PD-L1 and S100A9 levels in CD14 + monocytes from patients with HCC, CRC, and BTC. i S100A9 and CD274 ( j ) mRNA levels of S100A9 -knockdown THP-1 cells transfected with si-S100A9#1, si- S100A9 #2, and si- S100A9 #3. k Representative flow cytometric plot (left) and quantification (right) of S100A9 levels in S100A9 -knockdown THP-1 cells transfected with sh- S100A9 #1 and sh- S100A9 #2. l Representative flow cytometric plot (left) and quantification (right) of PD-L1 levels in S100A9 -knockdown THP-1 cells transfected with sh- S100A9 #1 and sh- S100A9 #2. m the percentage of highly proliferated CFSE low of T cells co-cultured with S100A9 -knockdown THP-1 cells at 96 h ( n = 3). n TBX21 , PRF1 , IL2 , and GZMB mRNA levels of T cells after co-cultured with S100A9 -knockdown THP-1 cells for 24 h. P value in ( b ) was evaluated using the Mann–whitney U-test. P values in c-e were determined by two-tailed paired sample t-test. P values in i-m were determined by one-way ANOVA. P value in ( n ) was evaluated using 2-way ANOVA. Correlations were analyzed using the Spearman rank correlation test. * P < 0.05, ** P < 0·01, *** P < 0.001 and **** P < 0.0001
Article Snippet: Staining was carried out using a 1:200 dilution of rabbit polyclonal antibody for human S100A9 (Proteintech Cat# 26,992–1-AP; RRID: AB_2880716) and a 1:2000 dilution of rabbit polyclonal antibody against human CD14 (Proteintech Cat# 17,000–1-AP; RRID: AB_2074048), followed by incubation with secondary antibodies (Biolynx Cat# I20012C).
Techniques: Expressing, Knockdown, Transfection, Cell Culture, MANN-WHITNEY, Two Tailed Test