Journal: Advanced Science

Article Title: Integrated Single‐Cell and TCR Profiling Reveals Protection‐Associated CD8 + T Cell Subsets Linked to Viral Control in PRRSV

doi: 10.1002/advs.76732

Figure Lengend Snippet: CD8 + T cells preferentially receive signals from macrophages /monocytes for activation via TLR4 signaling. (A) Communications between APC and CD8 + T cells within each experimental group as inferred by CellPhoneDB. (B) Analysis of the expression level of antigen presentation‐related genes in indicated cell subclusters. (C) Dot plot showing normalized mean expression level of genes involved in innate immunity in indicated subclusters of macrophages and monocytes. (D) The KEGG enrichment analysis of innate immunity‐associated pathways for differentially upregulated genes in the subclusters of macrophages and monocytes. The enrichment level is determined by the number of included genes and their expression levels. (E) Percentage of cells containing virus RNA fragments in the indicated cell clusters. (F, G) Role of TLR4 signaling in T cell activation. The spleen cell mixture from three pigs of JXA1R groups at 5dpc were prepared. The ability to secret IFN‐γ was determined using ELISpot in the presence of TAK‐242, a TLR4 inhibitor or absence of macrophage/monocytes that were depleted by mouse monoclonal antibodies to CD14. The representative ELISpot wells (F) and statistical analysis (G) were presented. (H, I) Role of TLR8 signaling in T cell activation with TLR8 inhibitor. The representative ELISpot wells (H) and statistical analysis (I) were presented. Statistical analysis was performed by two‐tailed Student's t‐test, and error bars indicate means ± standard error of mean (SEM). Asterisks (*) indicate the statistical significance: *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: For the TLR‐4 or TLR‐8 loss‐function assay, the CD14‐deleted cells were pre‐incubated with TAK‐242 (MCE, #243984‐11‐4) or CU‐CPT8m (MCE, #HY‐112050) at a final concentration of 5 μM for 2 h, followed by the addition of virus, and ELISpot was then performed.

Techniques: Activation Assay, Expressing, Immunopeptidomics, Virus, Enzyme-linked Immunospot, Bioprocessing, Two Tailed Test

Journal: Advanced Science

Article Title: Integrated Single‐Cell and TCR Profiling Reveals Protection‐Associated CD8 + T Cell Subsets Linked to Viral Control in PRRSV

doi: 10.1002/advs.76732

Figure Lengend Snippet: Schematic model summarizing immune features associated with protection and pathogenesis. In the protected group, antigen presentation is associated with specialized subset of macrophages and monocytes characterized by high TLR4 and pro‐inflammatory signals expression. With appropriate CD4 + T cell help, these CD8 + T cells acquire robust cytotoxic effector functions and mediate efficient antiviral immunity. In contrast, in unprotected animals, antigen presentation is associated with monocyte populations expressing suppressive and inhibitory signals. CD8 + T cells in this context exhibit increased expression of exhaustion‐associated markers, especially CTLA4 , together with reduced effector functionality. Aberrant CD4 + T cell–CD8 + T cell communication further reinforces T cell exhaustion, ultimately resulting in impaired antiviral responses and disease progression.

Article Snippet: For the TLR‐4 or TLR‐8 loss‐function assay, the CD14‐deleted cells were pre‐incubated with TAK‐242 (MCE, #243984‐11‐4) or CU‐CPT8m (MCE, #HY‐112050) at a final concentration of 5 μM for 2 h, followed by the addition of virus, and ELISpot was then performed.

Techniques: Immunopeptidomics, Expressing, Biomarker Discovery