Journal: Cell Reports Medicine
Article Title: Spleen-targeted neoantigen mRNA vaccine induces ISG15 + CD8 + T cell-mediated tertiary lymphoid structure formation in hepatocellular carcinoma
doi: 10.1016/j.xcrm.2026.102754
Figure Lengend Snippet: Spatial localization of ISG15 + CD8 + T cells and APCs and the formation of TLS following STNvac treatment (A–D) Spatial transcriptomics analysis of tumor samples isolated from PBS- and STNvac-treated mice. (A) H&E staining and corresponding spatial clustering of transcriptome spots; 19 clusters were identified, with clusters 4 and 7 showing marked enrichment of immune cells (CD45 + ). (B) Spatial expression of Ptprc (CD45) mapped on H&E-stained tissues, highlighting immune-cell-dense regions. (C) Region of interest annotation based on TESLA, illustrating colocalization of ISG15 + CD8 + T cells (approximated by Pclaf / Birc5 ), B cells, DCs, and CD4 + T cells; CXCL13 expression overlapped with these immune clusters, and TLS-like regions were identified using a TLS score (co-localization of B cells, CD4 + T cells, DCs, and CXCL13). (D) Magnified views of regions I and II from (A), showing compact lymphoid aggregates (white arrows) and smaller, loosely organized foci (black arrows) at the tumor margin corresponding to TLS. (E–G) Multicolor immunofluorescence staining analysis of TLSs and ISG15 + CD8 + T cells in tumors from different groups. (E) Representative images showing CD20 + B cells and CD23 + follicular regions within TLSs, together with ISG15 + CD8 + T cells localized around TLS structures. Scale bars: 500 μm; inset, 50 μm. (F) Magnified views of Region I from (E), including H&E and additional multicolor staining for CD20, CD21, CD23, and CD34, highlighting organized FDC networks and HEV-like microvessels within TLS boundaries. Scale bars, 100 μm (left) and 50 μm (right). (G) Quantitative analysis of TLS density in tumors from different treatment groups ( n = 3 biological replicates). (H) Spatial transcriptomic mapping of human HCC tissues from Liu et al.’s study showing ISG15 + CD8 + T cells located near or within TLS-like regions in immune-inflamed tumors. See also .
Article Snippet: TCGA: LIHC dataset was obtained from GDC Data Portal and patients were stratified according to the ssGSEA scores of the signature containing CD8A, CD8B and ISG15, the survival analysis was then further conducted using survival packages in R. Mendeley Data: http://www.doi.org/10.17632/skrx2fz79n.1 (the spatial transcriptome dataset from Liu’s research ) was downloaded with the link provided in that publication.
Techniques: Spatial Transcriptomics, Isolation, Staining, Expressing, Multicolor Immunofluorescence Staining