Journal: Clinical and Translational Medicine

Article Title: Single‐cell transcriptome sequencing of B‐cell heterogeneity and tertiary lymphoid structure predicts breast cancer prognosis and neoadjuvant therapy efficacy

doi: 10.1002/ctm2.1346

Figure Lengend Snippet: Correlation analysis between TIL‐B and mature tertiary lymphoid structures (TLS). (A) Among the 200 patients included in the BRCA dataset from the TCGA research consortium, a subset of 18 patients who exhibited TLS positivity were identified. H&E images of characteristic TLS in these patients. (B) Co‐immunofluorescence subimages depicting the co‐localization of CD23 and CD21 within CD23‐positive TLS were obtained from a collection of 70 BC samples that we compiled. (C) In 9 TLS‐positive patients of 70 BC patients, representative images of H&E staining and IHC staining of CD4, CD8, CD20, CD23 and BCL6 expression in TLS‐positive BC patients. (D) Analysis of DEGs in 200 TCGA BRCA cases categorized as TLS‐negative and TLS‐positive. Source data are available at: https://portal.gdc.cancer.gov . (E and F) H&E (E) and CD23 IHC (F) images of the typical TLS‐positive case in 70 BC patients. (G) Different tumour types exhibit FCER2 expression, according to TIMER2. * p < .05;** p < .01;*** p < .001. (H) Utilizing the EPIC and TIDE algorithms within TIMER2, the relationship between FCER2 expression and tumour infiltrating immune cells in BC was computed. (I‐J) Kaplan–Meier Survival curve of FCER2(I), co‐acting with FCER2 and B cells(J) in BC by TIMER2. TIL‐B, tumor‐infiltrating B cells.

Article Snippet: 1% bovine serum albumin for 10 min at room temperature, followed by overnight incubation at 4°C with the following primary antibodies: monoclonal antibody against FCER2 (CD23; 1:400; 60208‐2‐Ig; Proteintech), anti‐CD21 (1:400; 24374‐1‐AP; Proteintech), anti‐CD20 (1:400; 60271‐1‐Ig; Proteintech), anti‐CD4 (1:400; 67786‐1‐Ig; Proteintech), anti‐CD8 (1:400; 66868‐1‐Ig; Proteintech) and anti‐BCL6 (1:400; 66340‐1‐Ig; Proteintech).

Techniques: Immunofluorescence, Staining, Immunohistochemistry, Expressing

Journal: Clinical and Translational Medicine

Article Title: Single‐cell transcriptome sequencing of B‐cell heterogeneity and tertiary lymphoid structure predicts breast cancer prognosis and neoadjuvant therapy efficacy

doi: 10.1002/ctm2.1346

Figure Lengend Snippet: Different tertiary lymphoid structures (TLS) states in breast cancer (BC) display metabolic variability and survival variations. (A) A bar graph depicting the distribution of immune cells that have infiltrated tumours in TLS‐positive and TLS‐negative groups among 9 BC patients. (B) Genetic analysis of immune cell differences in TLS‐positive and TLS‐negative groups among 9 BC patients. (C) Heatmap of metabolic pathway scores in TLS‐positive and TLS‐negative groups among 9 BC patients. (D) Heatmap showing average metabolic gene expression in TLS‐positive and TLS‐negative groups among 9 BC patients. (E) To ascertain the expression correlation between CD23 and the biomarkers citrate synthase (CS), HK2, lactate dehydrogenase A (LDHA), and IDH3G in BC, the cBioportal tool ( https://www.cbioportal.org ) was utilized, employing the TCGA dataset (Firehose Legacy dataset; encompassing 1108 BC samples) for this purpose. (F) Kaplan–Meier survival curves depicting the disparity in BC outcomes between cohorts with high and low APOD expression, as analyzed through GEPIA. (G) Imaging before and after neoadjuvant chemotherapy of TLS‐positive patients with significant effectiveness. (H) Imaging before and after neoadjuvant immunotherapy of TLS‐positive patients with significant effectiveness.

Article Snippet: 1% bovine serum albumin for 10 min at room temperature, followed by overnight incubation at 4°C with the following primary antibodies: monoclonal antibody against FCER2 (CD23; 1:400; 60208‐2‐Ig; Proteintech), anti‐CD21 (1:400; 24374‐1‐AP; Proteintech), anti‐CD20 (1:400; 60271‐1‐Ig; Proteintech), anti‐CD4 (1:400; 67786‐1‐Ig; Proteintech), anti‐CD8 (1:400; 66868‐1‐Ig; Proteintech) and anti‐BCL6 (1:400; 66340‐1‐Ig; Proteintech).

Techniques: Gene Expression, Expressing, Imaging

Journal: Clinical and Translational Medicine

Article Title: Single‐cell transcriptome sequencing of B‐cell heterogeneity and tertiary lymphoid structure predicts breast cancer prognosis and neoadjuvant therapy efficacy

doi: 10.1002/ctm2.1346

Figure Lengend Snippet: The potential significance of tertiary lymphoid structures (TLS) and TLS‐specific markers for breast cancer (BC) prognosis. After matching with Fitness score matching (PSM), survival analysis and prognostic factor analysis were performed. For survival analysis, the Kaplan–Meier method and the log‐rank test were employed. For prognostic factors, COX proportional hazard regression model was used. The two‐sided log rank test was used to determine the p values. (A) Kaplan–Meier survival curves for the disease‐free survival (DFS) (left) and OS (right) of 920 TCGA BRCA patients based on single gene expression (CD20, CD23 and CD8). (B) The impact of CD20, CD23, CD8, CD4 and BCL6 on 920 TCGA BRCA patients’ prognosis. The forest map shows HRs (center pink and blue squares) and 95% confidence interval (horizontal ranges), and PSM matching has been made for factors such as molecular typing, lymph node status, tumour size, diagnosis age and histological grading of BC. (C) Kaplan–Meier survival curves for the DFS of 70 BC patients based on TLS expression.

Article Snippet: 1% bovine serum albumin for 10 min at room temperature, followed by overnight incubation at 4°C with the following primary antibodies: monoclonal antibody against FCER2 (CD23; 1:400; 60208‐2‐Ig; Proteintech), anti‐CD21 (1:400; 24374‐1‐AP; Proteintech), anti‐CD20 (1:400; 60271‐1‐Ig; Proteintech), anti‐CD4 (1:400; 67786‐1‐Ig; Proteintech), anti‐CD8 (1:400; 66868‐1‐Ig; Proteintech) and anti‐BCL6 (1:400; 66340‐1‐Ig; Proteintech).

Techniques: Gene Expression, Biomarker Discovery, Expressing