Targeted Delivery of Nanovaccine to Dendritic Cells via DC-Binding Peptides Induces Potent Antiviral Immunity in vivo.

Background: Dendritic cell (DC) targeted antigen delivery is a promising strategy to enhance vaccine efficacy and delivery of therapeutics. Self-assembling peptide-based nanoparticles and virus-like particles (VLPs) have attracted extensive interest as non-replicating vectors for nanovaccine design, based on their unique properties, including molecular specificity, biodegradability and biocompatibility. DCs are specialized antigen-presenting cells involved in antigen capture, processing, and presentation to initiate adaptive immune responses. Using DC-specific ligands for targeted delivery of antigens to DCs may be utilized as a promising strategy to drive efficient and strong immune responses. Methods: In this study, several candidates for DC-binding peptides (DCbps) were individually integrated into C-terminal of porcine circovirus type 2 (PCV2) Cap, a viral protein that could self-assemble into icosahedral VLPs with 60 subunits. The immunostimulatory adjuvant activity of DC-targeted VLPs was further evaluated in a vaccine model of PCV2 Cap. Results: With transmission electron microscopy (TEM), E. coli expressed Cap-DCbp fusion proteins were observed self-assembled into highly ordered VLPs. Further, in dynamic light scattering (DLS) analysis, chimeric VLPs exhibited similar particle size uniformity and narrow size distribution as compared to wild type Cap VLPs. With a distinctly higher targeting efficiency, DCbp3 integrated Cap VLPs (Cap-DCbp3) displayed enhanced antigen uptake and increased elicitation of antigen presentation-related factors in BM-DCs. Mice subcutaneously immunized with Cap-DCbp3 VLPs exhibited significantly higher levels of Cap-specific antibodies, neutralizing antibodies and intracellular cytokines than those with other DCbp integrated or wild type Cap VLPs without any DCbp. Interestingly, Cap-DCbp3 VLPs vaccine induces robust cellular immune response profile, including the efficient production of IFN-γ, IL-2 and IL-10. Meanwhile, the improved proliferation index in lymphocytes with Cap-DCbp3 was also detected as compared to other VLPs. Conclusion: This study described the potential of DC-binding peptides for further improved antigen delivery and vaccine efficacy, explainning nanovaccine optimization in relation to a range of emerging and circulating infectious pathogens.

Self-Assembling Nanovaccine Enhances Protective Efficacy Against CSFV in Pigs.

Classical swine fever virus (CSFV) is a highly contagious pathogen, which pose continuous threat to the swine industry. Though most attenuated vaccines are effective, they fail to serologically distinguish between infected and vaccinated animals, hindering CSFV eradication. Beneficially, nanoparticles (NPs)-based vaccines resemble natural viruses in size and antigen structure, and offer an alternative tool to circumvent these limitations. Using self-assembling NPs as multimerization platforms provides a safe and immunogenic tool against infectious diseases. This study presented a novel strategy to display CSFV E2 glycoprotein on the surface of genetically engineered self-assembling NPs. Eukaryotic E2-fused protein (SP-E2-mi3) could self-assemble into uniform NPs as indicated in transmission electron microscope (TEM) and dynamic light scattering (DLS). SP-E2-mi3 NPs showed high stability at room temperature. This NP-based immunization resulted in enhanced antigen uptake and up-regulated production of immunostimulatory cytokines in antigen presenting cells (APCs). Moreover, the protective efficacy of SP-E2-mi3 NPs was evaluated in pigs. SP-E2-mi3 NPs significantly improved both humoral and cellular immunity, especially as indicated by the elevated CSFV-specific IFN-γ cellular immunity and >10-fold neutralizing antibodies as compared to monomeric E2. These observations were consistent to in vivo protection against CSFV lethal virus challenge in prime-boost immunization schedule. Further results revealed single dose of 10 µg of SP-E2-mi3 NPs provided considerable clinical protection against lethal virus challenge. In conclusion, these findings demonstrated that this NP-based technology has potential to enhance the potency of subunit vaccine, paving ways for nanovaccine development.

MiR-30c-5p suppresses migration, invasion and epithelial to mesenchymal transition of gastric cancer via targeting MTA1.

BACKGROUND: In China, gastric cancer (GC) is an ordinary malignant tumor. Recent literatures have shown that microRNA is critical during tumorigenesis. This study focuses on the influence of miR-30c-5p on the metastasis of GC and further explores its underlying mechanism. METHODS: Before the study, expression level of miR-30c-5p and targeted protein was detected in 40 GC tissue samples and 5 GC cells by RT-qPCR. Meanwhile, correlation analysis was conducted between miR-30c-5p expression level and clinicopathological features. In addition, wound healing assay and cell invasion assay were utilized to identify whether miR-30c-5p could affect the migrated and invaded ability of GC cells. Western blotting assay and luciferase assay were used to explore the potential mechanism. RESULTS: In GC tissues, miR-30c-5p expression level was significantly lower and was remarkably related with clinical features such as tumor node metastasis(TNM) stage and lymphatic metastasis. Moreover, the migrated and invaded ability of GC cells was enhanced through knockdown of miR-30c-5p, while overexpression of miR-30c-5p presented with reversed effect. Further study showed that miR-30c-5p inhibited the expression of its target spot, metastasis-associated protein 1(MTA1), and then suppressed the process of epithelial to mesenchymal transition(EMT) which was important in the metastasis of GC. CONCLUSION: The results indicate that miR-30c-5p, a novel suppressor in tumorigenesis, could inhibit the metastasis and EMT via MTA1, which may offer a possible therapeutic target in GC.

The potential signal pathway between PAX2 and CD2AP in the renal interstitial fibrosis disease.

Paired box gene 2 (PAX2) can regulate tissue development and cellular differentiation, and it is associated with renal diseases. CD2-associated protein (CD2AP) is an adaptor protein involving in a variety of physiological and disease processes. Renal interstitial fibrosis (RIF) is a hallmark of common progressive chronic diseases which lead to renal failure. This study was performed to investigate whether there was a potential signal pathway between PAX2 and CD2AP in RIF rats induced by unilateral ureteral obstruction (UUO). Eighty Wistar male rats were divided into two groups randomly: sham operation group (SHO) and model group subjected to UUO (GU), n = 40. The model was established by left ureteral ligation. Renal tissues were collected at 14 d and 28 d after surgery. RIF index, cell apoptosis index, protein expression of PAX2, CD2AP, transforming growth factor-ßl (TGF-ß1), collagen-IV (Col-IV), fibronectin (FN) in renal interstitium and renal tissue, and mRNA expression of PAX2, CD2AP, and TGF-ß1 in renal tissue were detected. Compared with that in the SHO group, the PAX2 and CD2AP expressions (mRNA and protein) were significantly increased (p < 0.01). Protein expressions of TGF-ß1, Col-IV, and FN, and RIF index or cell apoptosis index in the GU group were markedly elevated than those in the SHO group (all p < 0.01). PAX2 or CD2AP was positively correlated with TGF-ß1, Col-IV, and FN, and RIF index or cell apoptosis index (all p < 0.05). Furthermore, PAX2 was positively correlated with CD2AP (p < 0.05). In conclusion, the expression of PAX2 or CD2AP was increased in RIF rats, and PAX2 was positively correlated with CD2AP. There might be a potential signaling pathway between PAX2 and CD2AP in RIF disease. Further research is needed to determine the association in RIF disease.

LIM homeobox transcription factor 1B is associated with pro-fibrotic components and apoptosis in hypoxia/reoxygenation renal tubular epithelial cells.

LIM homeobox transcription factor 1B (LMX1B) is a transcription factor of the LIM-homeodomain type, which plays an important role in foetal development during formation of the extremities, kidneys, eyes, and the brain. Furthermore, LMX1B has been implicated in nail-patella syndrome, which is predominantly characterized by malformation of limbs and nails, and in 30 % of patients, nephropathy, including renal fibrosis, is observed. Since no reports were available that studied the link between LMX1B expression and pro-fibrotic components and apoptosis in hypoxic renal tubular epithelial cells (RTEC), we explored if LMX1B was associated with extracellular matrix components, profibrotic factors, and apoptosis induced by hypoxia/reoxygenation (H-R). In this cell system under hypoxic conditions, when the expression of LMX1B was inhibited in H-R RTEC, the expression of transforming growth factor-ßl, collagen-III, fibronectin, cleaved caspase-3, and cell apoptosis rate was increased. Consequently, overexpression of LMX1B was associated with reduced cell apoptosis, whilst downregulation of LMX1B was associated with increased cell apoptosis in H-R RTEC.

All-trans retinoic acid attenuates the renal interstitial fibrosis lesion in rats but not by transforming growth factor-ß1/Smad3 signaling pathway.

All-trans retinoic acid (ATRA) is an important therapeutic agent for prevention of the renal diseases. Transforming growth factor-ß1 (TGF-ß1)/Smad3 signaling pathway is a key signaling pathway which takes part in the progression of renal interstitial fibrosis (RIF). This investigation was performed to study the effect of ATRA in RIF rats and its effect on the TGF-ß1/Smad3 signaling pathway. Sixty Wistar male rats were divided into three groups at random: sham operation group (SHO), model group subjected to unilateral ureteral obstruction (GU), model group treated with ATRA (GA), n = 20, respectively. RIF index, protein expression of TGF-ß1, collagen-IV (Col-IV) and fibronectin (FN) in renal interstitium, and mRNA and protein expressions of Smad3 in renal tissue were detected at 14-day and 28-day after surgery. The RIF index was markedly elevated in group GU than in SHO group (p < 0.01), and the RIF index of GA group was alleviated when compared with that in GU group (p < 0.01). Compared with in group SHO, the mRNA/protein expression of Smad3 in renal tissue was significantly increased in group GU (p < 0.01). However, the mRNA and protein expressions of Smad3 in renal tissue in GA group were not markedly alleviated by ATRA treatment when compared with those in GU (each p > 0.05). Protein expressions of TGF-ß1, Col-IV, and FN in GU group were markedly increased than those in SHO group (each p < 0.01), and their expressions in GA group were markedly down-regulated by ATRA treatment than those of GU group (all p < 0.01). The protein expression of Smad3 was positively correlated with RIF index, protein expression of TGF-ß1, Col-IV or FN (each p < 0.01). In conclusion, ATRA treatment can alleviate the RIF progression in UUO rats. However, ATRA cannot affect the signaling pathway of TGF-ß1/Smad3 in the progression of RIF.

Zhou, T.B., et Al., correction: all-trans retinoic Acid treatment is associated with prohibitin expression in renal interstitial fibrosis rats. Int. J. Mol. Sci. 2012, 13, 2769-2782.

The authors wish to change Figure 2 of the paper published in IJMS [1]. The positions of H(1) and H(2) in the previous article were reversed. These errors have been amended in an amended version of the manuscript, which is available from the International Journal of Molecular Sciences website. The authors and publisher apologize for the inconvenience. [...].

All-trans retinoic acid treatment is associated with prohibitin expression in renal interstitial fibrosis rats.

This study was performed to investigate the association of prohibitin with renal interstitial fibrosis (RIF) lesion and to explore the association of all-trans retinoic acid (ATRA) treatment with prohibitin expression in RIF rats. Rats were divided into three groups: the sham operation group (SHO), the model group subjected to unilateral ureteral obstruction (UUO), and the model group treated with ATRA (GA). Renal tissues were collected at 14 and 28 days after surgery, and the relevant indicators were detected. In comparison with the SHO group, the RIF index in the UUO group was markedly elevated (p < 0.01), and the RIF index in the GA group was alleviated compared with that in the UUO group (p < 0.01). Compared with the SHO group, the expression of prohibitin (protein or mRNA) in the UUO group was significantly reduced (each p < 0.01). Prohibitin expression in the GA group was markedly increased when compared with that in the UUO (p < 0.01). The expression of TGF-ß1 (protein and mRNA), protein expressions of Col-IV, fibronectin, α-SMA and cleaved Caspase-3, ROS generation and cell apoptosis index in the UUO group were markedly higher than those in the SHO group (all p < 0.01), and their expressions in the GA group were markedly down-regulated compared to those in the UUO group (all p < 0.01, respectively). The protein expression of prohibitin was negatively correlated with the RIF index, protein expression of TGF-ß1, Col-IV, fibronectin, α-SMA or cleaved Caspase-3, ROS generation and the cell apoptosis index (each p < 0.01). In conclusion, lower expression of prohibitin is associated with the RIF, and ATRA treatment is associated with increased prohibitin, which can prevent the progression of RIF.