rat anti dc lamp  (Novus Biologicals)

Journal: Nature Communications

Article Title: Mature tertiary lymphoid structures evoke intra-tumoral T and B cell responses via progenitor exhausted CD4 + T cells in head and neck cancer

doi: 10.1038/s41467-025-59341-w

Figure Lengend Snippet: a Heatmap plot showing TLS status in HNSCC (TCGA-HNSC) database ( n = 500 independent samples) identified by the different immune cell types. A color gradient, transitioning from purple (representing enrichment) to white (representing depletion). The Benjamini–Hochberg adjusted P values obtained from two-sided Kruskal–Wallis tests. b Overall survival of patients with HNSCC by TLS status ( n = 499 independent samples). Overall survival of patients with HNSCC by GC B cells ( c ) and CD8 + effector memory T cells ( d ) was analyzed using log-rank test results, which indicated statistical significance. e Multivariate Cox proportional regression outcome, with clinical variables. For every variable included in the analysis, the reference level is the first one. A gray bar symbolizes a P value > 0.05; and blue and red bars symbolize P value < 0.05 positively and negatively, respectively. Error bars represent the 95% confidence interval. f Scheme of mIHC for HNSCC tissue. g – l Proportion of CD20 + , CD4 + , CD8 + , DC-LAMP + , CD8 + TCF1 + , and CD4 + TCF1 + CXCL13 + cells in total cells of HNSCC tissue with different TLS status, respectively ( n = 303 independent samples nTLS, n = 83 independent samples imTLS, n = 36 independent samples mTLS, two-tailed Mann–Whitney U -test, for box plots: box center line, median; box limits, upper and lower quartiles; box whiskers, maximum and minimum values). m Heatmap showing the Pearson correlation among cell subcluster abundances. The color gradient indicates the P value of Pearson correlation (red, high expression; white, low expression). n Images of an mIHC-stained HNSCC tissue with different statuses of TLS showing co-localization of B cell (CD20), T cell (CD4, CD8, and TCF1), mregDC (DC-LAMP), and chemokine (CXCL13) in HNSCC across. nTLS were repeated 303 times independently with similar results, imTLS were repeated 83 times independently with similar results, mTLS were repeated 36 times independently with similar results. Scale bars = 200 μm. o The Distance between each type of cell in each core of the TMAs, with cores grouped by TLS status. Vertical dotted lines indicate the median distance. p Relationship between differential prioritization ∆Augur score for parenting between response and no response, and Augur score for HNSCC patients treated with ICB therapy. The scRNA-seq data of pre-treatment tumor samples from six patients with HNSCC undergoing ICB therapy GSE200996 were used for Augur analysis. Each dot represents a cell type in the TME of HNSCC. Gray symbolizes a ∆Augur score of <0.02; and the colored dot symbolizes positively and negatively a ∆Augur score’s absolute value of >0.02, respectively. q Heatmap plot showing the response to anti-PD-1 therapy and abundance of different immune cell types. GSE93157 was used for this analysis. The Benjamini–Hochberg adjusted P values obtained from a two-tailed Mann–Whitney U -test. r Sketch map showing the TME of HNSCC with nTLS, imTLS, and mTLS. nTLS are unable to generate local anti-tumor immunity due to a lack of stem-like T cells and B cells. Meanwhile, imTLS, which is also enriched with B cells, cannot produce in situ anti-tumor immunity due to obstruction by immunosuppressive cells, including Tregs and M2 macrophages. mTLS enriched with stem-like CD4 + and CD8 + T cells and the presence of B cells at different maturation stages, supporting the generation of T and B cell responses. CD4 + Tex prog /Tfh collaborated with cDC2 and mregDCs play a crucial role in regulating B cell and T cell response within mTLS. CR complete response, PR partial response, SD stable disease, PD progressive disease.

Article Snippet: Briefly, FFPE tissue slides were first deparaffinized and then incubated sequentially with primary antibodies CD20 (1:1000, Cell Signaling Technology), CD4 (1:1000, Abcam), CD8 (1:800, Cell Signaling Technology), CXCL13 (1:600, Abcam), DC-LAMP (1:600, Atlas Antibodies), TCF1 (1:600, Cell Signaling Technology), PD-1 (1:600, Cell Signaling Technology), CCR7 (1:1200, Abcam), CXCR5 (1:1200, Abcam), Granzyme B (1:800, Cell Signaling Technology), BCL6 (1:600, Cell Signaling Technology), CD23 (1:900, Abcam) and CD1C (1:1000, Abcam).

Techniques: Two Tailed Test, MANN-WHITNEY, Expressing, Staining, In Situ