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Additional file 1: Figure S1. Characterization of phenotype and differentiation potential of hucMSCs. A. Morphology of cultured hucMSCs. (B-I) Surface antigen expression on hucMSCs by flow cytometry. Cells were stained with antibodies against mouse IgG1 isotype FITC (B), mouse IgG1 isotype PE (C), mouse IgG1 isotype APC (D), CD90 FITC (E), CD73 PE (F), CD105 APC (G), CD45 FITC (H), CD34 PE (I). (J-L) Differentiation potential of hucMSCs into adipocytes (J), chondrocytes (K), and osteocytes (L). Figure S2. Gating strategy of lung fibroblast. (A) Representative gating strategy of EpCAM-CD31-CD45- lung fibroblast. Figure S3. In vivo imaging of DiR-labeled hucMSCs distribution. (A) Control mice (saline-treated) and bleomycin-treated mice were injected with DiR-labeled hucMSCs at day 0, and imaged at day 1, day 3, day 5, day 7, and day 14. Figure S4. Macrophage markers expression on a t-SNE plot. (A) Normalized expression of macrophage markers Adgre1, Cd68, and Lyz2 overlaid on a t-SNE plot. Figure S5. Dynamics of lung macrophage after bleomycin treatment. (A) Representative gating strategy of macrophages. (B) Quantification of lung macrophages in saline-treated mice lungs, bleomycin-treated mice lungs, and hucMSCs-treated mice lungs at day 7 and day 14. (C) Representative gating strategy of alveolar macrophage. (D) Quantification of alveolar macrophages in saline-treated mice lungs, bleomycin-treated mice lungs, and hucMSCs-treated mice lungs at day 7 and day 14 (mean ± SD, n = 3 mice per group). (E) Representative gating strategy of interstitial macrophages and monocytes. (F) Quantification of interstitial macrophages and monocytes in saline-treated mice lungs, bleomycin-treated mice lungs, and hucMSCs-treated mice lungs at day 7 and day 14 (mean ± SD, n = 3 mice per group). *p < 0.05, ***p < 0.001, n.s. no significant difference, Student’s t-test.

Placenta-specific 1 (PLAC1), a novel cancer-testis antigen (CTA), is expressed in a number of different human malignancies. It is frequently produced in breast cancer, serving a function in tumorigenesis. Adoptive immunotherapy using T cell receptor (TCR)-engineered T cells against CTA mediates objective tumor regression; however, to the best of our knowledge, targeting PLAC1 using engineered T cells has not yet been attempted. In the present study, the cDNAs encoding TCRα- and β-chains specific for human leukocyte antigen (HLA)-A*0201-restricted PLAC1 were cloned from a cytotoxic T-lymphocyte, generated by in vitro by the stimulation of CD8+ T cells using autologous HLA-A2+ dendritic cells loaded with a PLAC1-specific peptide (p28-36, VLCSIDWFM). The TCRα/β-chains were linked by a 2A peptide linker (TCRα-Thosea asigna virus-TCRβ), and the constructs were cloned into the lentiviral vector, followed by transduction into human cytotoxic (CD8+) T cells. The efficiency of transduction was up to 25.16%, as detected by PLAC1 multimers. TCR-transduced CD8+ T cells, co-cultured with human non-metastatic breast cancer MCF-7 cells (PLAC1+, HLA-A2+) and triple-negative breast cancer MDAMB-231 cells (PLAC1+, HLA-A2+), produced interferon γ and tumor necrosis factor α, suggesting TCR activation. Furthermore, the PLAC1 TCR-transduced CD8+ T cells efficiently and specifically identified and annihilated the HLA-A2+/PLAC1+ breast cancer cell lines in a lactate dehydrogenase activity assay. Western blot analysis demonstrated that TCR transduction stimulated the production of mitogen-activated protein kinase signaling molecules, extracellular signal-regulated kinases 1/2 and nuclear factor-κB, through phosphoinositide 3-kinase γ-mediated phosphorylation of protein kinase B in CD8+ T cells. Xenograft mouse assays revealed that PLAC1 TCR-transduced CD8+T cells significantly delayed the tumor progression in mice-bearing breast cancer compared with normal saline or negative control-transduced groups. In conclusion, a novel HLA-A2-restricted and PLAC1-specific TCR was identified. The present study demonstrated PLAC1 to be a potential target for breast cancer treatment; and the usage of PLAC1-specific TCR-engineered T cells may be a novel strategy for PLAC1-positive breast cancer treatment.

Target molecules selectively equipped with proximity probes can autonomously cleave substrates on the electrode surface, allowing quantification of proteins at attomolar concentrations with one-step incubation., Various strategies have been proposed for the detection of disease protein biomarkers; however, most methods are too expensive, cumbersome or limited in sensitivity for clinical use. Here, we report that a fabricated complex can be used as a powerful tool to detect trace proteins in complex samples. In this strategy, a DNA–protein complex that comprises of one target molecule and two or more deoxyribozyme-containing probes can exhibit autonomous cleavage behavior on the surface of the substrate DNA modified electrode. In the meantime, the complex can remove the cleaved DNA fragment from the electrode surface by taking advantage of the proximity effect. The proposed approach allows one-step and highly sensitive detection of a variety of targets based on the changes of the direct electrochemical readout. Moreover, this method may also have considerable advantages over the commonly reported DNA amplification-assisted immunoassays, particularly in terms of assay simplicity and cost, which may hold great potential for application in resource-constrained regions.

Background aims Acute liver failure (ALF), a life-threatening disease characterized by the sudden loss of hepatic function, can occur after an accidental or intentional acetaminophen overdose. Methods With the use of an ALF mouse model, we examined both the preventive and therapeutic potential of intravenously administered human umbilical cord–derived mesenchymal stromal cells (hUCMSCs). Primary hUCMSCs were purified from freshly collected full-term umbilical cords and intravenously transplanted into BALB/c mice either before and after ALF induced by acetaminophen intoxication. We found that hUCMSCs significantly improved survival rates and relative liver weight of mice in both pre-ALF and post-ALF animals. Correspondingly, serum levels of markers that reflect hepatic injury (ie, aspartate aminotransferase, alanine aminotransferase and total bilirubin) were significantly attenuated in the group receiving hUCMSC therapy. Results Mechanistically, we found that the protective potential of intravenously administered hUCMSCs was mediated by paracrine pathways that involved antioxidants (glutathione, superoxide dismutase), the reduction of inflammatory agents (tumor necrosis factor-α, interleukin-6) and elevated serum levels of hepatocyte growth factor. Conclusions Through these paracrine effects, intravenously administered hUCMSCs reduced hepatic necrosis/apoptosis and enhanced liver regeneration. Thus, our data demonstrate that intravenously administered hUCMSCs may be useful in the prevention or treatment of acetaminophen-induced ALF.

Background aims Acute radiation syndrome (ARS) leads to pancytopenia and multi-organ failure. Transplantation of hematopoietic stem cells provides a curative option for radiation-induced aplasia, but this therapy is limited by donor availability. Methods We examined an alternative therapeutic approach to ARS with the use of human extracellular superoxide dismutase (ECSOD)-modified umbilical cord mesenchymal stromal cells (UCMSCs). This treatment combines the unique regenerative role of UCMSCs with the anti-oxidative activity of ECSOD. Results We demonstrated that systemically administered ECSOD-UCMSCs are able to protect mice from sub-lethal doses of radiation and improve survival by promoting multilineage hematopoietic recovery. The therapeutic effect of this treatment is related to the decrease in radiation-induced O 2 – and apoptosis. Conclusions Our data highlight the clinical potential of this two-pronged approach to the treatment of ARS, thereby serving as a rapid and effective first-line strategy to combat the hematopoietic failure resulting from a radiation accident, nuclear terrorism and other radiologic emergencies.