Fig 2: The antitumour effect of LIA to inhibit primary and abscopal tumour growth.a Schematic illustration of LIA-mediated antitumour effect to inhibit primary tumour and abscopal tumour growth. b, c Tumour volumes of 4T1 primary tumours (b) and abscopal tumours (c) were evaluated (n = 8). d, Images of the 4T1 primary tumours and abscopal tumours from different groups at day 24. 1, 2, 3, 4, 5 and 6 represent tumour images from mice treated with PBS, Laser, LIA, Free Ce6+L, DP-Ce6+L and LIA+L at an equivalent Ce6 dose of 3 mg/kg, respectively, scale bar is 1 cm. e–j T-cell infiltration was analysed 7 days after the final treatment. e–g Flow cytometry data of CD3+ CD8+ cell proportion in T cells in abscopal tumours (e), spleens (f) and CD4+ CD25+ foxp3+ cell proportion in CD4+ T cells (g). e–g experiments were performed three times independently, representative images are shown. h–j Statistical analysis of CD3+ CD8+ cell proportion in T cells in abscopal tumours (h), spleens (i) and CD4+ CD25+ foxp3+ cell proportion in CD4+ T cells (j). k, l Antitumour efficiency tested on CT26 bilateral tumour model. Tumour volumes of CT26 primary tumours (k) and abscopal tumours (l) were evaluated (n = 8). m Representative immunofluorescence images of CD8+ (green) and CD4+ (red) cells infiltrated into abscopal tumours in CT26 tumour model with different treatments. Scale bar, 50 µm. Experiments in m were performed three times independently, representative images are shown. Data in b, c and k, l are presented as mean ± s.d. (n = 8). Data in h, i, j are presented as mean ± s.d. (n = 3). Statistically significant differences between groups were identified by one-way ANOVA. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05.

Fig 3: Study design. (A): Protocol of allergic airway inflammation induction. OP administration of PBS (naïve) or AHE solution on days 0 and 7 to initiate the immune response, then challenged for 3 days on days 14, 15, and16 with OP inoculations of AHE. Cell administration was performed on day 14; harvest was on day 19. (B): Experimental groups. Animals exposed to AHE solution (A) were randomly divided to receive treatment with the vehicle PBS, HLFs, mMSCs, BMDMCs, and BMDMCs depleted of CD45, CD34, CD11b, CD3, CD19, CD31, and Sca-1 positive cells. Abbreviations: #, oropharyngeal Aspergillus hyphal extract solution administered; ?, human lung fibroblasts, mouse mesenchymal stem/stromal cells, bone marrow-derived mononuclear cells and depleted bone marrow-derived mononuclear cells administered; A, animals exposed to AHE solution; AHE, Aspergillus hyphal extract; BMDMC, bone marrow-derived mononuclear cell; HLF, human lung fibroblast; mMSC, mouse mesenchymal stem/stromal cell; OP, oropharyngeal; P, group receiving phosphate-buffered saline; PBS, phosphate-buffered saline.

Fig 4: Donor mLNs are repopulated with recipient hematopoietic cells after surgical transplantation. (A) Flow cytometry gating of hematopoietic cells in mLNs from B6.CD45.1 mouse transplanted in a B6.CD45.2 mouse. (B) Absolute numbers of hematopoietic cells, myeloid cell (dendritic cells and macrophages), B cells and CD3+ pre-gated CD4+ and CD8+ T cells in CD45.1+ and CD45.2+ sub-populations, 5 weeks after mLN transplantation. Data pooled from five mice analyzed in two experiments. (C) Flow cytometry gating of non-hematopoietic LNSCs and DsRed fluorescence signal from B6.DsRed mLNs transplanted in a B6.WT mouse, with DsRed+ living cells from a DsRed+ mouse as positive and living cells from a WT mouse serving as negative controls. Analysis performed 5 weeks after mLN transplantation. Error bars represent Mean± SD.

Fig 5: Bone-marrow mononuclear cell characterization before and after specific Dynabead depletions. Percentage of CD45-, CD34-, CD3-, CD19-, CD11b-, CD31-, and Sca-1-positive cells evaluated by flow cytometry analysis of bone-marrow derived mononuclear cells before and after magnetic depletions. Representative depletions are shown. Data are presented as mean + SD (n = 3). *, significance compared with BMDMC before depletion.