Fig 2: Lung cDC2 and cDC1 DCs present MHC class I epitope SIINFEKL and are activated after intranasal SIIN-Q11(A) Ability of CD11c+ DCs and CD45− cells enriched from the lungs of naive or SIIN-Q11 immunized mice to stimulate B3Z reporter cells. B3Z cells recognize and respond to SIINFEKL:Kb complexes with the upregulation of LacZ.(B)Representative flow cytometry plots of lung SIINFEKL:Kb presenting cDC1 and cDC2 DCs at 2 days post intranasal SIIN-Q11 immunization. cDCs were identified as CD45+CD49b−TER119−CD19−CD3−SiglecF−Ly6G−CD11c+MHCIIhiCD64−cells, and SIINFEKL:H-2Kb-positive cDCs were detected using 25-D1.16 mAb.(C) Groups for (D)–(H).(D) Number of lung SIINFEKL:Kb presenting cDC1 and cDC2 DCs at day 2 post intranasal SIIN-Q11 immunization.(E–G) CD80 (E), CD86 (F), and CD40 (G) mean fluorescence intensity (MFI) on lung cDC1 and cDC2 at 2 days post intranasal nanofibers immunization. Each dot represents the number of SIINFEKL:Kb-presenting lung DCs from one mouse. Data are means ± SEM from two independent experiments (A and D–G). ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05; ns, not significant by unpaired two-tailed t test.(H) PCA of RNA-seq from the four DC groups.(I) Heatmap of DEGs and k-means clustering of DEGs based on pairwise comparison of four DC groups using DESeq2 (n = 580; adjusted p <0.01). The color key for Z score, and the three main clusters (S-Q11:SIIN-Q11).(J) Boxplots of the Z scores of cluster 1 and 2 DEGs according to experimental groups. Significant differences in Z scores between the indicated DC groups were assessed by one-way ANOVA. ****p < 0.0001; ns, not significant.(K) Bar charts of Metascape enrichment analysis of DEGs from clusters 1 (top) and 2 (bottom).

Fig 3: 68Ga-FAPI micro-PET/CT and 18F-FDG micro-PET/CT scans to assess tumor response to combined therapy with TGF-βR inhibitor and ICB KN046 in mice with colorectal peritoneal metastasis.(A) Schematic of micro-MRI and PET imaging and treatment strategies in mice with MC38 peritoneal metastasis (4 groups, n = 6 per group). Created with BioRender. (B) Representative micro-MRI images of mice with peritoneal metastasis after the indicated treatments. Yellow arrows indicate tumor lesions. (C) Tumor weight of mice with MC38 peritoneal metastasis after the indicated treatments. (D–F) Proportion of CD8+ T cells, CD8+IFN-γ+ T cells, and CD8+GZMB+ T cells in CD45+CD3+ cells in peritoneal metastasis tumors harvested from mice in the 4 groups as determined using flow cytometry. (G) Quantified tumor uptake of 68Ga-FAPI in mice with peritoneal metastasis (n = 6 per group). (H) Representative 68Ga-FAPI micro-PET/CT images of mice with peritoneal metastasis after the indicated treatments. B, bladder; K, kidney; T, tumor. (I) Quantified tumor uptake of 18F-FDG in mice with peritoneal metastasis (n = 6 per group). (J) Representative 18F-FDG micro-PET/CT images of mice with peritoneal metastasis after the indicated treatments. (K) Quantified IHC staining of FAP-α in the tumors of mice with peritoneal metastasis after the indicated treatments. (L) Representative IHC staining of FAP-α in the tumors of mice with peritoneal metastasis after the indicated treatments. Scale bars: 20 μm. All numerical data are presented as mean ± SEM. *P < 0.05, **P < 0.01, *** P < 0.001, by 1-way ANOVA with Dunnett’s correct multiple-comparison test (C–G, I, and K).

Fig 4: DSF increases the CD4 + CD25+ Foxp3+ regulatory T cell subset in grafts.a Flow cytometry plot of CD25+Foxp3+ cells in the CD3 + CD4+ leukocyte gate of the graft-infiltrated cells. b Data of FCM profiles of the percentage of CD25+ Foxp3+ cells in CD4+ cells (left) and absolute cell number CD4 + CD25+ Foxp3+ cells (right). n = 7 biologically independent mice. c Data of FCM profiles of the percentage of CD4 + T cells in leukocytes (left) and absolute cell number CD4 + T cells (right). n = 7 biologically independent mice. d Immunofluorescent staining of CD4+ (green) and Foxp3+ (red) cells in the graft. Significance was calculated using two-tailed unpaired Student’s t test. *P < 0.05, **P < 0.01.

Fig 5: Intravenous delivery of glunomab reduces neuroinflammation and DA-neuron degeneration and rescues behavioral deficits after overexpression of hα-SYN. Representative images of the SN stained for (A) TH and (C) MHC-I, and the respective quantification of (B) DA-neuron survival and (D) MHC-I fluorescence intensity fold change in the SN 4 weeks after rAAV2-hα-SYN injection in WT mice treated with glunomab (10 mg/kg/week) or vehicle (n = 6–10). (E) Confocal image of the SN showing CD3+ cells (red) in contact with TH+ cell bodies and axons. (F) Quantification of CD3+ T cells associated with the SNpc 4 weeks after rAAV2-hα-SYN in WT mice treated with glunomab or vehicle (n = 7–10). (G) Quantification of sensorimotor bias in a corridor task 4 weeks after rAAV2-hα-SYN in WT mice treated with glunomab (10 mg/kg/week) or vehicle (n = 9–10). Glunomab treatment was performed on 12- and 16-week-old WT mice. For vehicle treatment, age-matched 12- and 16-week-old WT mice were used. Data are shown as mean ± SEM, N.S = not significant, *p < 0.05; ****p < 0.0001. (B, F, G) 2-tailed t-test; (D) 1-way ANOVA followed by Tukey post hoc test. Scale bar= (A, C) 250 μm (E) 20 μm