cd14 microbeads miltenyi biotec  (Miltenyi Biotec)

Journal: Oncoimmunology

Article Title: Ex vivo-generated conventional dendritic cells type 1 and type 2 from blood progenitors induce potent antigen-specific T-cell immunity

doi: 10.1080/2162402X.2026.2695692

Figure Lengend Snippet: Ex vivo-generated cDC1s and cDC2s acquire a matured state upon TLR challenge. Enriched cDC1s and cDC2s were overnight stimulated with a maturation cocktail containing R848 and Poly(IC), and the next day, their phenotype and cytokine/chemokine production were characterized. (A) Representative histograms showing the expression levels of CD83, CD40, and CCR7 upon maturation on DC2s (blue histograms) and cDC1s (red histograms). (B) Maturation marker expression on cDC2s (blue dots) and cDC1s (red dots) is displayed as either percentage of positive DCs (CD80, CD86, CD83, CD40, and CCR7) or relative geometric mean fluorescence intensity (gMFI) (HLA-DR). Relative gMFI expression was determined by normalizing to the average gMFI of untreated cDC1s or cDC2s. Scatter dot plots show the mean ± SEM with each data point representing an individual DC donor ( n ≥ 8). (C) Scatter dot plots show the mean ± SEM of the raw concentrations of the indicated cytokines and chemokines. Each data point represents an individual DC donor ( n ≥ 8). Statistical significance was calculated for paired data sets with a paired t-test or a Wilcoxon test, whereas for unpaired data sets significancy was calculated using a Mann–Whitney test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: After 2 h of coculture at 37oC, cells were stained to identify DC2s (CD1c-FITC, 1:25, 130-113-301, Miltenyi Biotec) and cDC1s (CD141 PerCP-Cy5.5, 1:25, 344112, BioLegend) prior to analysis using a BD FACSLyricTM.

Techniques: Ex Vivo, Generated, Expressing, Marker, Fluorescence, MANN-WHITNEY

Journal: Oncoimmunology

Article Title: Ex vivo-generated conventional dendritic cells type 1 and type 2 from blood progenitors induce potent antigen-specific T-cell immunity

doi: 10.1080/2162402X.2026.2695692

Figure Lengend Snippet: CD34-derived cDC1s and cDC2s can be generated from blood progenitors. CD34-positive cells isolated from blood were differentiated into cDC1s and cDC2s in a 17-day protocol in which progenitors initially expand and afterward undergo skewing towards cDC1 and cDC2 differentiation. (A) Schematic representation of the 17-day protocol for DC generation. (B) Bar graph showing the percentage of cDC1s and cDC2s ± SEM at the end of the culture on day 17 ( n = 10). (C) Representative dot plot showing the presence of cDC1s and cDC2s based on the expression of CD141/CLEC9A and CD1c, respectively, after gating on viable single cells. (D) Violin plot showing the absolute number of cDC1s and cDC2s obtained with the current protocol, depending on the initial amount of cultured CD34-positive cells. Each dot represents the value of expanded cDC1s and cDC2s obtained from one independent donor ( n = 12). (E) Bar graphs show the percentage of positive DCs ± SEM for the given marker ( n = 3). CD34-derived cDC1s and cDC2s were characterized by flow cytometry to determine their lineage marker expression.

Article Snippet: Alternatively, DCs were enriched by MACS using CD141-Microbeads (130-090-512, Miltenyi Biotec) and CD1c Isolation kit (130-119-475, Miltenyi Biotec) sequentially (see supplementary figure 1C).

Techniques: Derivative Assay, Generated, Isolation, Expressing, Cell Culture, Marker, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Tumor immune microenvironment states inferred from TLS-associated immune-cell composition stratify prognosis in hepatocellular carcinoma

doi: 10.3389/fimmu.2026.1818138

Figure Lengend Snippet: TLS RiskScore association with response to PD-1 blockade and protein-level implementation in FFPE HCC. (A, B) External anti–PD-1-treated HCC cohort: TLS RiskScore by response status (A) and response rates by TLS RiskScore group using the median cutoff (B) . In (B) , colors indicate the within-group proportions of responders and non-responders within each TLS RiskScore group. (C) Spatial transcriptomics mapping canonical TLS-associated chemokines (CXCL13, CCL19, CCL21) and signature signals for regulatory T cells (Treg) and conventional type 2 dendritic cells (cDC2) in representative HCC sections. (D, E) Representative TLS regions in FFPE HCC illustrating cDC2-predominant versus Treg-predominant cellular composition by H&E and multiplex immunofluorescence (mIF) staining (CD11c, CD1c, CD4, FOXP3, CD45, CD21). (F, G) Overall survival (OS) (F) and progression-free survival (PFS) (G) stratified by an mIF-based TLS RiskScore derived from TLS-region Treg (CD4 + FOXP3 + ) and cDC2 (CD11c + CD1c + ) measurements (median cutoff). (H) cDC2 fractions across TLS maturation stages. (I) Treg-to-cDC2 ratio across TLS maturation stages. **P < 0.01 and ****P < 0.0001.

Article Snippet: The antibody panel included CD45 (Aifang, AF20018), CD4 (Aifang, AF20210), FOXP3 (Maxim, MAB-1004), CD11c (Aifang, AF20381), CD21 (Aifang, AF20013), and CD1c (HUABIO, HA722562), each used at 1:200.

Techniques: Spatial Transcriptomics, Multiplex Assay, Immunofluorescence, Staining, Derivative Assay