Genetic fusion of CCL11 to antigens enhances antigenicity in nucleic acid vaccines and eradicates tumor mass through optimizing T-cell response.

Nucleic acid vaccines have shown promising potency and efficacy for cancer treatment with robust and specific T-cell responses. Improving the immunogenicity of delivered antigens helps to extend therapeutic efficacy and reduce dose-dependent toxicity. Here, we systematically evaluated chemokine-fused HPV16 E6/E7 antigen to improve the cellular and humoral immune responses induced by nucleotide vaccines in vivo. We found that fusion with different chemokines shifted the nature of the immune response against the antigens. Although a number of chemokines were able to amplify specific CD8 + T-cell or humoral response alone or simultaneously. CCL11 was identified as the most potent chemokine in improving immunogenicity, promoting specific CD8 + T-cell stemness and generating tumor rejection. Fusing CCL11 with E6/E7 antigen as a therapeutic DNA vaccine significantly improved treatment effectiveness and caused eradication of established large tumors in 92% tumor-bearing mice (n = 25). Fusion antigens with CCL11 expanded the TCR diversity of specific T cells and induced the infiltration of activated specific T cells, neutrophils, macrophages and dendritic cells (DCs) into the tumor, which created a comprehensive immune microenvironment lethal to tumor. Combination of the DNA vaccine with anti-CTLA4 treatment further enhanced the therapeutic effect. In addition, CCL11 could also be used for mRNA vaccine design. To summarize, CCL11 might be a potent T cell enhancer against cancer.

Transient Inhibition of the JNK Pathway Promotes Human Hematopoietic Stem Cell Quiescence and Engraftment.

The widespread clinical application of cord blood (CB) for hematopoietic stem cell (HSC) transplantation is limited mainly by the inadequate number of hematopoietic stem and progenitor cells (HSPCs) in single CB units, which results in unsuccessful or delayed engraftment in recipients. The identification of agents to promote CB HSPC engraftment has significant therapeutic value. Here, we found that transient inhibition of the JNK pathway increased the HSC frequency in CB CD34+ cells to 13.46-fold. Mechanistic studies showed that inhibition of the JNK pathway upregulated the expression of quiescence-associated and stemness genes in HSCs, preventing HSCs from entering the cell cycle, increasing glucose uptake and accumulating reactive oxygen species (ROS). Importantly, transient inhibition of the JNK pathway during CB CD34+ cell collection also enhanced long-term HSC (LT-HSC) recovery and engraftment efficiency. Collectively, these findings suggest that transient inhibition of the JNK pathway could promote a quiescent state in HSCs by preventing cell cycle entry and metabolic activation, thus enhancing the HSC number and engraftment potential. Together, these findings improve the understanding of the regulatory mechanisms governing HSC quiescence and stemness and have the potential to improve HSC collection and transplantation.

Efficacy and Safety of Central Memory T Cells Combined With Adjuvant Therapy to Prevent Recurrence of Hepatocellular Carcinoma With Microvascular Invasion: A Pilot Study.

BACKGROUND: Postoperative adjuvant transcatheter arterial chemoembolization (TACE) following curative hepatectomy has been reported to improve the clinical outcomes of hepatocellular carcinoma (HCC) patients with microvascular invasion (MVI), but more endeavors are required to achieve greater clinical benefit. Central memory T-cell (Tcm) self-transfusion has shown superior antitumor activity in several preclinical studies; however, clinical studies are rare. The aim of this study was to evaluate the clinical benefit and safety of combination treatment with Tcm self-transfusion and TACE as adjuvant treatment in HCC patients with MVI after curative hepatectomy. METHODS: From October 2016 to September 2018, primary HCC patients with histologically confirmed MVI who underwent curative hepatectomy at the Cancer Hospital of the Chinese Academy of Medical Sciences were recruited for this study. The patients were divided into a Tcm group (combined Tcm self-transfusion with TACE treatment) or a control group (TACE treatment alone) according to their willingness. The recurrence-free survival (RFS), quality-of-life (QOL) score, and adverse events of each patient were recorded within 2 years. RESULTS: A total of 52 patients were enrolled, and 48 were eligible for the final data analysis. The median follow-up time was 20.5 months (95% CI: 17.05-22.55 months). The median RFS time was 9.5 months in the control group; the cutoff date was not reached in the Tcm group (when the follow-up duration was 12 months, p = 0.049, HR = 0.40; 95% CI: 0.16-0.99). Compared with the control group, 1- and 2-year RFS rates were higher in the Tcm group (72.0% vs. 46.4% and 58.18% vs. 39.14%, respectively). Multivariate analysis did not indicate that Tcm treatment was an independent prognostic factor associated with HCC recurrence (p = 0.107, HR = 2.312; 95% CI: 0.835-6.400), which might be due to the small sample size of this study. Nevertheless, Tcm treatment effectively improved a reduced QOL due to HCC and liver function injury. Finally, the safety profile of Tcm treatment in this study was good, without any serious adverse events. CONCLUSIONS: This pilot study showed that Tcm self-transfusion combined with TACE treatment might be a beneficial adjuvant therapy with good safety for primary HCC patients with MVI after curative hepatectomy. TRIAL REGISTRATION NUMBER: NCT03575806.

Gold-Nanoparticle-Functionalized Cobalt-Nickel Phosphate 3D Nanoice Creams to Fabricate Stable and Sensitive Biosensors for the Cytochrome c Assay.

Designing a stable and sensitive luminol-based electrochemiluminescence (ECL) analytical platform in the neutral condition has attracted a lot of attention. Here, gold-nanoparticle-functionalized cobalt-nickel phosphate three-dimensional nanoice creams (Au@Co3Ni3(PO4)4 NICs) are successfully prepared via electrostatic interaction. Generally, cobalt-nickel phosphate nanoice creams (Co3Ni3(PO4)4 NICs) are synthesized via a mild hydrothermal method and functionalized via polyethylenimine (PEI). Then, Au NPs are adsorbed on the surface of Co3Ni3(PO4)4 NICs via Au-N weak interaction to fabricate Au@Co3Ni3(PO4)4 NICs. Owing to the important roles of Au@Co3Ni3(PO4)4 in exhibiting excellent electrocatalytic activity, as well as preventing the deposition of negatively charged oxidation product induced electrode passivation, luminol in the nanohybrids (LH-Au@Co3Ni3(PO4)4) gives strong and stable ECL intensity in the neutral conditions. Moreover, the ECL emission of luminol is obviously quenched based on the resonance energy transfer (RET) between luminol as donor and cytochrome c (Cyt c) as acceptor. Hence, a sensitive ECL biosensor is successfully fabricated for the quantitative determination of Cyt c in cell lysates and exhibits wide linear ranges of 1.0 × 10-4-0.5 × 10-5 and 0.5 × 10-5-1.0 × 10-8 M as well as a low detection limit of 2.48 nM. This novel sensing strategy will broaden the application of transition metal (Co, Ni) phosphates in bioassays.

A novel ultrasensitive electrochemiluminescence biosensor for glutathione detection based on poly-L-lysine as co-reactant and graphene-based poly(luminol/aniline) as nanoprobes.

In this work, an ultrasensitive electrochemiluminescence (ECL) biosensor was constructed using poly-L-lysine (PLL) as a novel co-reactant of luminol and poly(luminol/aniline) nanorods loaded reduced graphene oxide (PLA@rGO) as nanoprobe, which enable highly sensitivity detection of glutathione (GSH). To the best of our knowledge, it is the first time that PLL was used for the co-reactant of luminol. Notably, about a 5-fold enhancement was obtained compared with the individual PLA@rGO using GCE. Due to the remarkable quenching effect between the excited state of PLL and the reduced form of GSH in the ECL system of luminol/PLL, the ECL sensing platform exhibited wide linear ranges of 1.0 × 10-9-1.0 × 10-4 M and 1.0 × 10-4-1.0 × 10-2 M and a low detection limit of 7.7 × 10-10 M. Simultaneously, the biosensor was also successfully applied to detect GSH in human serum sample with high recoveries. Hence, this work would open a new platform for the wide application of PLL in immunoassay and various sensors.

The rice "fruit-weight 2.2-like" gene family member OsFWL4 is involved in the translocation of cadmium from roots to shoots.

MAIN CONCLUSION: Heterogeneous expression of the rice genes "fruit-weight 2.2-like" (OsFWL) affects Cd resistance in yeast, and OsFWL4 mediates the translocation of Cd from roots to shoots. Cadmium (Cd) induces chronic and toxic effects in humans. In a previous study (Xu et al. in Planta 238:643-655, 2013), we cloned the rice genes, designated OsFWL1-8, homologous to the tomato fruit-weight 2.2. Here, we show that expression of genes OsFWL3-7 in yeast confers resistance to Cd. The Cd contents of OsFWL3-, -4-, -6- and -7-transformed Cd(II)-sensitive yeast mutant ycf1 cells were strongly decreased compared with those of empty vector, with the strongest resistance to Cd observed in cells expressing OsFWL4. Evaluation of truncated and site-directed mutation derivatives revealed that the CCXXG motifs near the second transmembrane region of OsFWL4 are involved in Cd resistance in yeast. Real-time PCR analysis showed that OsFWL4 expression was induced by CdCl2 stress in rice seedlings. Compared with WT plants, the Cd contents in the shoots of RNAi mediated OsFWL4 knockdown plants were significantly decreased, and Cd translocation from roots to shoots was reduced. According to bimolecular fluorescence complementation, yeast two-hybrid and Western-blotting assays, the OsFWL4 protein forms homo-oligomers. These results suggest that OsFWL4 might act directly as a transporter and is involved in the translocation of Cd from roots to shoots in rice.

Direct Reprogramming of Fibroblasts via a Chemically Induced XEN-like State.

Direct lineage reprogramming, including with small molecules, has emerged as a promising approach for generating desired cell types. We recently found that during chemical induction of induced pluripotent stem cells (iPSCs) from mouse fibroblasts, cells pass through an extra-embryonic endoderm (XEN)-like state. Here, we show that these chemically induced XEN-like cells can also be induced to directly reprogram into functional neurons, bypassing the pluripotent state. The induced neurons possess neuron-specific expression profiles, form functional synapses in culture, and further mature after transplantation into the adult mouse brain. Using similar principles, we were also able to induce hepatocyte-like cells from the XEN-like cells. Cells in the induced XEN-like state were readily expandable over at least 20 passages and retained genome stability and lineage specification potential. Our study therefore establishes a multifunctional route for chemical lineage reprogramming and may provide a platform for generating a diverse range of cell types via application of this expandable XEN-like state.

Effect of Sanhuangyilong decoction plus methotrexate on tumor necrosis factor alpha and interferon gamma in serum and synovial fluid in rheumatoid arthritis patients with symptom pattern of damp heat obstruction.

OBJECTIVE: To investigate the effect of Sanhuangyilong decoction plus methotrexate (MTX) on Interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) in the serum and synovial fluid of rheumatoid arthritis (RA) patients with damp-heat-obstruction symptom pattern, Sanhuangyilong decoction and the role of TNF-α and IFN-γ in the development of RA. METHODS: RA inpatients with damp-heat-obstruction symptom pattern (partly with knee joint effusion) were selected as the research subjects. Before the treatment, healthy subjects and osteoarthritis (OA) patients with knee joint effusion were assigned to the serum control group and the synovial fluid control group, respectively; during the treatment, RA patients with damp-heat-obstruction symptom pattern were divided into two groups: one is combined group that was administered Sanhuangyilong decoction plus MTX; the other group was MTX group that received MTX only. The expression levels of TNF-α and IFN-γ in the serum and synovial fluid were measured with enzyme-linked immunosorbent assay (ELISA) before and after the treatment, and the peripheral blood levels of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and disease activity score in 28 joints (DAS28) were determined. RESULTS: Before treatment, the serum levels of TNF-α and IFN-γ in the RA patients with dampheat- obstruction symptom pattern were higher than those in healthy control group (P < 0.05).The expression levels of TNF-α and IFN-γ in the synovial fluid of the RA patients were higher than those in the serum of the RA patients (P < 0.05). The expression levels of TNF-α and IFN-γ in the synovial fluid of the RA patients were higher than those of the synovial fluid of the osteoarthritis patients (P < 0.05). The expression of TNF-α and IFN-γ in the serum and synovial fluid of the RA patients had no correlation with the inflammatory activity index ESR, CRP, or DAS28 (P > 0.05). After 2 weeks of treatment, the expression level of TNF-α and IFN-γ in the combined group had increased, although the difference was not statistically significant (P > 0.05); in contrast, ESR, CRP, and DAS28 decreased, and the difference was statistically significant (P < 0.01). After 4 weeks of therapy, TNF-alpha and IFN-γ, ESR, CRP, and DAS28 in the combined group decreased compared with the before-treatment levels (P < 0.01). After 2 w of treatment, the differences in the TNF-α and IFN-γ expression levels in the combined group were not statistically significant (P > 0.05) compared with that in the MTX group, although there were statistically significant differences in the ESR, CRP, and DAS28 (P < 0.05). After 4 weeks of treatment, differences in TNF-α, IFN-γ, ESR, CRP, and DAS28 in the combined group compared with MTX group were statistically significant (P < 0.01). CONCLUSION: TNF-α and IFN-γ might be involved in the development of RA. The RA patients with damp-heat-obstruction symptom pattern show better benefits from the treatment of Sanhuangyilong decoction plus MTX, and the treatment is superior to that of using MTX only.