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

Journal: Journal of Translational Autoimmunity

Article Title: Type I interferon-dependent and -independent signaling underlie autoantibody production in a murine lupus model

doi: 10.1016/j.jtauto.2026.100351

Figure Lengend Snippet: Induction of SLE autoantibodies occurs in the presence of LPS or Poly I:C, but not other TLR ligands. Wild type (WT) C57BL/6 mice received five immunizations with β2GPI in combination with one of the following TLR ligands: TLR2 : PAM2, FSL-1 (lipoprotein); TLR3 : Poly I:C (dsRNA); TLR4 : LPS; TLR7: IMQ (imiquimod); or TLR9 : CpG-ODN (ssDNA). Serum autoantibodies were detected by ELISA. Bars indicate the mean value +/standard error (S.E.) per group (n = 5 mice/group), and dots indicate values for individual mice. Autoantibody levels for control mice immunized with PBS and TLR ligand were OD 405 < 0.1. The OD 405 + 3 S.D. for unimmunized mice was ≤0.1 for all autoantibodies assayed. (A) Anti-β2GPI and anti-CL antibodies, and (B) hallmark SLE autoantibodies (anti-DNA, anti-Ro/SS-A, anti-La/SS-B, and anti-Sm) were analyzed statistically in separate groupings. For each of the two autoantibody categories, each different TLR ligand (immunized with β2GPI) was compared to the other TLR ligands by 2-way ANOVA, followed by a Tukey's multiple comparison test. ns = not significant. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001.

Article Snippet: TLR ligands Pam2CSK4 (PAM2), FSL-1, Pam3CSK4 (PAM3), Poly I:C (HMW), imiquimod-R837 (IMQ), and CpG-ODN-1555 + 1466 (CpG-ODN) were from InvivoGen (San Diego, CA).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Comparison

Journal: bioRxiv

Article Title: Liver sinusoidal endothelial cells integrate metabolic and immune signals for MAPK-dependent BMP6 regulation and hepcidin induction

doi: 10.64898/2026.05.07.723498

Figure Lengend Snippet: (A) Bmp6 mRNA expression in primary LSECs treated with 5 ng/mL LPS or 2.5 μM heme for 6 h in the presence or absence of hepatocyte-conditioned medium. (B) Bmp6 mRNA expression in primary LSECs treated with various TLR ligands: Pam3CSK4 (TLR1/2), PGN (TLR2), Poly I:C (TLR3), LPS (TLR4), FLA-ST (TLR5), FSL1 (TLR2/6), R848 (TLR7/8), and ODN (TLR9) or vehicle control (NT, non-treated) for 6 h. (C-D) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of LPS for 6 h or with 5 ng/mL LPS for 2, 4, and 6 h. (E) Bmp6 mRNA expression in LSECs treated with 2.5 μM heme or protoporphyrin IX (PPIX) for 6 h. (F-G) Bmp6 mRNA expression in primary LSECs treated with increasing concentrations of heme for 6 h or treated with 2.5 μM heme for 2, 4, or 6 h. (H) Bmp6 mRNA expression in primary LSECs pre-treated with TAK242 (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS or 2.5 μM heme treatment for 6 h. (I) Transcription factor activity analysis by RNA-seq of LSECs treated with heme with respect to untreated controls (contrast-wise), in presence of hepatocyte-conditioned medium. (J) Bmp6 mRNA expression in primary LSECs pre-treated with CHX (5 μM) or DMSO for 1 h, followed by 5 ng/mL LPS treatment for 6 h. Cell culture experiments, except those in panel A, were always conducted in the presence of hepatocyte-conditioned medium. Gene expression levels were assessed by RT-qPCR, normalized to the housekeeping gene Rpl19 , and expressed as fold change relative to vehicle-treated controls. The dashed line (ut) represents the mRNA expression of LSECs treated with the conditions shown, in the absence of LPS or heme. Data are obtained from three or four independent experiments and displayed as mean ± SD. Statistical significance: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, one-way ANOVA or Student’s t-test. CM, conditioned medium; PPIX, protoporphyrin IX; CHX, cycloheximide.

Article Snippet: The TLR ligands Pam3CSK4 (TLR2:1) (#tlrl-pms), PGN-SA (TLR2) (#tlrl-pgns2), Poly I:C (TLR3) (#tlrl-picw), FLA-ST (TLR5) (#tlrl-stfla), FSL1 (TLR2:TLR6) (tlrl-fsl), R848 (TLR7:8) (#tlrl-r848-1), ODN (TLR9) (#tlrl-1826) and the MAPK inhibitor SP600125 (#tlrl-sp60) were purchased form Invivogen and diluted in PBS (TLR ligands) or DMSO (SP600125).

Techniques: Expressing, Control, Activity Assay, RNA Sequencing, Cell Culture, Gene Expression, Quantitative RT-PCR