Plant‐expressed Fc‐fusion protein tetravalent dengue vaccine with inherent adjuvant properties

Kim, Mi Young,Copland, Alastair,Nayak, Kaustuv,Chandele, Anmol,Ahmed, Muhammad S.,Zhang, Qibo,Diogo, Gil R.,Paul, Matthew J.,Hofmann, Sven,Yang, Moon&#x2010,Sik,Jang, Yong&#x2010,Suk,Ma, Julian K&#x20 BLACKWELL 2018 PLANT BIOTECHNOLOGY JOURNAL Vol.16 No.7

<P><B>Summary</B></P><P>Dengue is a major global disease requiring improved treatment and prevention strategies. The recently licensed Sanofi Pasteur Dengvaxia vaccine does not protect children under the age of nine, and additional vaccine strategies are thus needed to halt this expanding global epidemic. Here, we employed a molecular engineering approach and plant expression to produce a humanized and highly immunogenic poly‐immunoglobulin G scaffold (PIGS) fused to the consensus dengue envelope protein III domain (cEDIII). The immunogenicity of this IgG Fc receptor‐targeted vaccine candidate was demonstrated in transgenic mice expressing human FcγRI/CD64, by induction of neutralizing antibodies and evidence of cell‐mediated immunity. Furthermore, these molecules were able to prime immune cells from human adenoid/tonsillar tissue <I>ex vivo</I> as evidenced by antigen‐specific CD4<SUP>+</SUP> and CD8<SUP>+</SUP> T‐cell proliferation, IFN‐γ and antibody production. The purified polymeric fraction of dengue PIGS (D‐PIGS) induced stronger immune activation than the monomeric form, suggesting a more efficient interaction with the low‐affinity Fcγ receptors on antigen‐presenting cells. These results show that the plant‐expressed D‐PIGS have the potential for translation towards a safe and easily scalable single antigen‐based tetravalent dengue vaccine.</P>

True Digestibility of Phosphorus in Different Resources of Feed Ingredients in Growing Pigs

Wu, X.,Ruan, Z.,Zhang, Y.G.,Hou, Y.Q.,Yin, Y.L.,Li, T.J.,Huang, R.L.,Chu, W.Y.,Kong, X.F.,Gao, B.,Chen, L.X. Asian Australasian Association of Animal Productio 2008 Animal Bioscience Vol.21 No.1

To determine the true digestible phosphorus (TDP) requirement of growing pigs, two experiments were designed with the experimental diets containing five true digestible P levels (0.16%, 0.20%, 0.23%, 0.26% and 0.39%) and the ratio of total calcium to true digestible P (TDP) kept at 2:1. In Experiment 1, five barrows (Duroc${\times}$Landrace${\times}$Yorkshire) with an average initial body weight of 27.9 kg were used in a $5{\times}5$ Latin-square design to evaluate the effect of different dietary P levels on the digestibility and output of P and nitrogen. In Experiment 2, sixty healthy growing pigs (Duroc${\times}$Landrace${\times}$Yorkshire) with an average body weight (BW) of 21.4 kg were assigned randomly to one of the five dietary treatments (12 pigs/diet), and were used to determine the true digestible phosphorus (TDP) requirement of growing pigs on the basis of growth performance and serum biochemical indices. The results indicated that the true digestibility of P increased (p<0.05) linearly with increasing dietary TDP level below 0.26%. The true P digestibility was highest (56.6%) when dietary TDP was 0.34%. Expressed as g/kg dry matter intake (DMI), fecal P output increased (p<0.05) linearly with increasing P input. On the basis of g/kg fecal dry matter (DM), fecal P output was lowest for Diet 4 and highest (p<0.05) for Diet 5. The apparent digestibility of crude protein (CP) did not differ (p>0.05) among the five diets, with the average nitrogen output of 12.14 g/d and nitrogen retention of 66% to 74% (p>0.05), which suggested that there was no interaction between dietary P and CP protein levels. During the 28-d experimental period of Experiment 2, the average daily gain (ADG) of pigs was affected by dietary TDP levels as described by Eq. (1): $y=-809,532x^4+788,079x^3-276,250x^2+42,114x-1,759$; ($R^2=0.99$; p<0.01; y = ADG, g/d; x = dietary TDP, %), F/G for pigs by Eq. (2): $y=3,651.1x^4-3,480.4x^3+1,183.8x^2-172.5x+10.9$ ($R^2=0.99$; p<0.01; y = F/G; x = dietary TDP, %), and Total P concentrations in serum by Eq. (3): $y=-3,311.7x^4+3,342.7x^3-1,224.6x^2+195.6x-8.7$ (R2 = 0.99; p<0.01; y = total serum P concentration and x = dietary TDP, %). The highest ADG (782 g/d), the lowest F/G (1.07) and the highest total serum P concentration (3.1 mmol/L) were obtained when dietary TDP level was 0.34%. Collectively, these results indicate that the optimal TDP requirement of growing pigs is 0.34% of the diet at a total Ca to TDP ratio of 2:1.

Potent Suppressive Effects of 1-Piperidinylimidazole Based Novel P2X7 Receptor Antagonists on Cancer Cell Migration and Invasion

Park, Jin-Hee,Williams, Darren R.,Lee, Ji-Hyung,Lee, So-Deok,Lee, Je-Heon,Ko, Hyojin,Lee, Ga-Eun,Kim, Sujin,Lee, Jeong-Min,Abdelrahman, Aliaa,Mu&#x308,ller, Christa E.,Jung, Da-Woon,Kim, Yong-Chul American Chemical Society 2016 Journal of medicinal chemistry Vol.59 No.16

<P>The P2X7 receptor (P2X7R) has been reported as a key mediator in inflammatory processes and cancer invasion/metastasis. In this study, we report the discovery of novel P2X7R antagonists and their functional activities as potential antimetastatic agents. Modifications of the hydantoin core-skeleton and the side chain substituents of the P2X7R antagonist 7 were performed. The structure activity relationships (SAR) and optimization demonstrated the importance of the sulfonyl group at the R-1 position and the substituted position and overall size of R-2 for P2X7R antagonism. The optimized novel analogues displayed potent P2X7 receptor antagonism (IC50 = 0.11-112 nM) along with significant suppressive effects on IL-1 beta release (IC50 = 0.32-210 nM). Moreover, representative antagonists (12g, 13k, and 17d) with imidazole and uracil core skeletons significantly inhibited the invasion of MDA-MB-231 triple negative breast cancer cells and cancer cell migration in a zebrafish xenograft model, suggesting the potential therapeutic application of these novel P2X7 antagonists to block metastatic cancer.</P>

Identification of a novel starfish neuropeptide that acts as a muscle relaxant

Kim, Chan&#x2010,Hee,Kim, Eun Jung,Go, Hye&#x2010,Jin,Oh, Hye Young,Lin, Ming,Elphick, Maurice R.,Park, Nam Gyu John Wiley and Sons Inc. 2016 Journal of Neurochemistry Vol.137 No.1

<P><B>Abstract</B></P><P>Neuropeptides that act as muscle relaxants have been identified in chordates and protostomian invertebrates but little is known about the molecular identity of neuropeptides that act as muscle relaxants in deuterostomian invertebrates (e.g. echinoderms) that are ‘evolutionary intermediates’ of chordates and protostomes. Here, we have used the apical muscle of the starfish <I>Patiria pectinifera</I> to assay for myorelaxants in extracts of this species. A hexadecapeptide with the amino acid sequence Phe‐Gly‐Lys‐Gly‐Gly‐Ala‐Tyr‐Asp‐Pro‐Leu‐Ser‐Ala‐Gly‐Phe‐Thr‐Asp was identified and designated starfish myorelaxant peptide (SMP). Cloning and sequencing of a cDNA encoding the SMP precursor protein revealed that it comprises 12 copies of SMP as well as 3 peptides (7 copies in total) that are structurally related to SMP. Analysis of the expression of SMP precursor transcripts in <I>P. pectinifera</I> using qPCR revealed the highest expression in the radial nerve cords and lower expression levels in a range of neuromuscular tissues, including the apical muscle, tube feet and cardiac stomach. Consistent with these findings, SMP also caused relaxation of tube foot and cardiac stomach preparations. Furthermore, SMP caused relaxation of apical muscle preparations from another starfish species – <I>Asterias amurensis</I>. Collectively, these data indicate that SMP has a general physiological role as a muscle relaxant in starfish. Interestingly, comparison of the sequence of the SMP precursor with known neuropeptide precursors revealed that SMP belongs to a bilaterian family of neuropeptides that include molluscan pedal peptides (PP) and arthropodan orcokinins (OK). This is the first study to determine the function of a PP/OK‐type peptide in a deuterostome.</P><P> Pedal peptide/orcokinin (PP/OK)‐type peptides are a family of structurally related neuropeptides that were first identified and functionally characterised in protostomian invertebrates. Here, we report the discovery of starfish myorelaxant peptide (SMP), a novel member of the PP/OK‐type neuropeptide identified in the starfish <I>Patiria pectinifera</I> (phylum Echinodermata). SMP is the first PP/OK‐type neuropeptide to be functionally characterised in a deuterostome.</P>

Charge‐Transport Anisotropy in a Uniaxially Aligned Diketopyrrolopyrrole‐Based Copolymer

Schott, Sam,Gann, Eliot,Thomsen, Lars,Jung, Seok&#x2010,Heon,Lee, Jin&#x2010,Kyun,McNeill, Christopher R.,Sirringhaus, Henning John Wiley and Sons Inc. 2015 Advanced Materials Vol.27 No.45

<P><B>Aligned films of a semiconducting DPP‐based copolymer</B> exhibit highly anisotropic charge transport with a band‐like temperature dependence along the alignment direction and hole mobilities of up to 6.7 cm<SUP>2</SUP> V<SUP>−1</SUP> s<SUP>−1</SUP>. X‐ray diffraction measurements reveal an exceptional degree of in‐plane alignment, high crystallinity, and a dominant face‐on orientation of the polymer backbones. The surprising charge‐transport properties are interpreted in a tie‐chain model consistent with anisotropic activation energies. </P>

Association of Obesity with Serum Leptin, Adiponectin, and Serotonin and Gut Microflora in Beagle Dogs

Park, H.&#x2010,J.,Lee, S.&#x2010,E.,Kim, H.&#x2010,B.,Isaacson, R.E.,Seo, K.&#x2010,W.,Song, K.&#x2010,H. John Wiley and Sons Inc. 2015 Journal of veterinary internal medicine Vol.29 No.1

<P><B>Background</B></P><P>Serotonin (5‐hydroxytryptamine, 5HT) is involved in hypothalamic regulation of energy consumption. Also, the gut microbiome can influence neuronal signaling to the brain through vagal afferent neurons. Therefore, serotonin concentrations in the central nervous system and the composition of the microbiota can be related to obesity.</P><P><B>Objective</B></P><P>To examine adipokine, and, serotonin concentrations, and the gut microbiota in lean dogs and dogs with experimentally induced obesity.</P><P><B>Animals</B></P><P>Fourteen healthy Beagle dogs were used in this study.</P><P><B>Methods</B></P><P>Seven Beagle dogs in the obese group were fed commercial food ad libitum, over a period of 6 months to increase their weight and seven Beagle dogs in lean group were fed a restricted amount of the same diet to maintain optimal body condition over a period of 6 months. Peripheral leptin, adiponectin, 5HT, and cerebrospinal fluid (CSF‐5HT) levels were measured by ELISA. Fecal samples were collected in lean and obese groups 6 months after obesity was induced. Targeted pyrosequencing of the 16S rRNA gene was performed using a Genome Sequencer FLX plus system.</P><P><B>Results</B></P><P>Leptin concentrations were higher in the obese group (1.98 ± 1.00) compared to those of the lean group (1.12 ± 0.07, <I>P</I> = .025). Adiponectin and 5‐hydroytryptamine of cerebrospinal fluid (CSF‐5HT) concentrations were higher in the lean group (27.1 ± 7.28) than in the obese group (14.4 ± 5.40, <I>P</I> = .018). Analysis of the microbiome revealed that the diversity of the microbial community was lower in the obese group. Microbes from the phylum Firmicutes (85%) were predominant group in the gut microbiota of lean dogs. However, bacteria from the phylum Proteobacteria (76%) were the predominant group in the gut microbiota of dogs in the obese group.</P><P><B>Conclusions and Clinical Importance</B></P><P>Decreased 5HT levels in obese group might increase the risk of obesity because of increased appetite. Microflora enriched with gram‐negative might be related with chronic inflammation status in obese dogs.</P>

Generation of anti‐tumour immune response using dendritic cells pulsed with carbonic anhydrase IX‐<i>Acinetobacter baumannii</i> outer membrane protein A fusion proteins against renal cell carcinoma

Kim, B.&#x2010,R.,Yang, E.&#x2010,K.,Kim, D.&#x2010,Y.,Kim, S.&#x2010,H.,Moon, D.&#x2010,C.,Lee, J.&#x2010,H.,Kim, H.&#x2010,J.,Lee, J.&#x2010,C. Blackwell Publishing Ltd 2012 Clinical and experimental immunology Vol.167 No.1

<P><B>Summary</B></P><P>Carbonic anhydrase IX (CA9), a specific molecular marker for renal cell carcinoma (RCC), serves as a potential target for RCC‐specific immunotherapy using dendritic cells (DCs). However, pulsing of DCs with CA9 alone is not sufficient for generation of a therapeutic anti‐tumour immune response against RCC. In this study, in order to generate a potent anti‐tumour immune response against RCC, we produced recombinant CA9‐<I>Acinetobacter baumannii</I> outer membrane protein A (AbOmpA) fusion proteins, designated CA9‐AbOmpA, and investigated the ability of DCs pulsed with CA9‐AbOmpA fusion proteins in a murine renal cell carcinoma (RENCA) model. A recombinant CA9‐AbOmpA fusion protein was composed of a unique proteoglycan‐related region of CA9 (1–120 amino acids) fused at the C‐terminus with transmembrane domain of AbOmpA (1–200 amino acids). This fusion protein was capable of inducing DC maturation and interleukin (IL)‐12 production in DCs. Interaction of DCs pulsed with CA9‐AbOmpA fusion proteins with naive T cells stimulated secretion of IL‐2, interferon (IFN)‐γ and tumour necrosis factor (TNF)‐α in T cells. Lymphocytes harvested from mice immunized with DCs pulsed with CA9‐AbOmpA fusion proteins secreted IFN‐γ and showed a specific cytotoxic activity against CA9‐expressing RENCA (RENCA‐CA9) cells. Administration of CA9‐AbOmpA‐pulsed DC vaccine suppressed growth of RENCA‐CA9 cells in mice with an established tumour burden. These results suggest that DCs pulsed with CA9‐AbOmpA fusion proteins generate a specific anti‐tumour immune response against RCC, which can be utilized in immunotherapy of RCC.</P>

A Distinct Role for Interleukin‐6 as a Major Mediator of Cellular Adjustment to an Altered Culture Condition

Son, Hwa&#x2010,Kyung,Park, Iha,Kim, Jue Young,Kim, Do Kyeong,Illeperuma, Rasika P.,Bae, Jung Yoon,Lee, Doo Young,Oh, Eun&#x2010,Sang,Jung, Da&#x2010,Woon,Williams, Darren R.,Kim, Jin John Wiley and Sons Inc. 2015 Journal of cellular biochemistry Vol.116 No.11

<P><B>ABSTRACT</B></P><P>Tissue microenvironment adjusts biological properties of different cells by modulating signaling pathways and cell to cell interactions. This study showed that epithelial–mesenchymal transition (EMT)/ mesenchymal–epithelial transition (MET) can be modulated by altering culture conditions. HPV E6/E7‐transfected immortalized oral keratinocytes (IHOK) cultured in different media displayed reversible EMT/MET accompanied by changes in cell phenotype, proliferation, gene expression at transcriptional, and translational level, and migratory and invasive activities. Cholera toxin, a major supplement to culture medium, was responsible for inducing the morphological and biological changes of IHOK. Cholera toxin per se induced EMT by triggering the secretion of interleukin 6 (IL‐6) from IHOK. We found IL‐6 to be a central molecule that modulates the reversibility of EMT based not only on the mRNA level but also on the level of secretion. Taken together, our results demonstrate that IL‐6, a cytokine whose transcription is activated by alterations in culture conditions, is a key molecule for regulating reversible EMT/MET. This study will contribute to understand one way of cellular adjustment for surviving in unfamiliar conditions. J. Cell. Biochem. 116: 2552–2562, 2015. © 2015 The Authors. <I>Journal of Cellular Biochemistry</I> Published by Wiley Periodicals, Inc.</P>

Brassinosteroid Biosynthesis Is Modulated via a Transcription Factor Cascade of COG1, PIF4, and PIF5

Wei, Zhuoyun,Yuan, Tong,Tarkowsks&#x30c,a&#x301,, Danus&#x30c,e,Kim, Jeongsik,Nam, Hong Gil,Novs&#x30c,a&#x301,r&#x30c,a&#x301,k, Onds&#x30c,a&#x301,r&#x30c,ej,He, Kai,Gou, Xiaoping,Li, Jia American Society of Plant Biologists 2017 Plant Physiology Vol.174 No.2

<P>Brassinosteroids (BRs) are essential phytohormones regulating various developmental and physiological processes during normal growth and development. cog1-3D (cogwheel1-3D) was identified as an activation-tagged genetic modifier of bri1-5, an intermediate BR receptor mutant in Arabidopsis (Arabidopsis thaliana). COG1 encodes a Dof-type transcription factor found previously to act as a negative regulator of the phytochrome signaling pathway. cog1-3D single mutants show an elongated hypocotyl phenotype under light conditions. A loss-of-function mutant or inducible expression of a dominant negative form of COG1 in the wild type results in an opposite phenotype. A BR profile assay indicated that BR levels are elevated in cog1-3D seedlings. Quantitative reverse transcription-polymerase chain reaction analyses showed that several key BR biosynthetic genes are significantly up-regulated in cog1-3D compared with those of the wild type. Two basic helix-loop-helix transcription factors, PIF4 and PIF5, were found to be transcriptionally up-regulated in cog1-3D. Genetic analysis indicated that PIF4 and PIF5 were required for COG1 to promote BR biosynthesis and hypocotyl elongation. Chromatin immunoprecipitation and electrophoretic mobility shift assays indicated that COG1 binds to the promoter regions of PIF4 and PIF5, and PIF4 and PIF5 bind to the promoter regions of key BR biosynthetic genes, such as DWF4 and BR6ox2, to directly promote their expression. These results demonstrated that COG1 regulates BR biosynthesis via up-regulating the transcription of PIF4 and PIF5.</P>

CD117⁺CD3⁻CD56⁻OX40L^high cells express IL‐22 and display an LTi phenotype in human secondary lymphoid tissues

Kim, Soochan,Han, Sinsuk,Withers, David R.,Gaspal, Fabrina,Bae, Jingyu,Baik, Song,Shin, Hyun&#x2010,Chool,Kim, Kyung&#x2010,Su,Bekiaris, Vasileios,Anderson, Graham,Lane, Peter,Kim, Mi&#x2010,Yeon WILEY‐VCH Verlag 2011 European journal of immunology Vol.41 No.6

<P><B>Abstract</B></P><P>Here, we identify cells within human adult secondary lymphoid tissues that are comparable in phenotype and location to the lymphoid tissue inducer (LTi) cells that persist in the adult mouse. Identified as CD117<SUP>+</SUP>CD3<SUP>−</SUP>CD56<SUP>−</SUP> cells, like murine LTi cells, they lack expression of many common lineage markers and express CD127, OX40L and TRANCE. These cells were detected at the interface between the B‐ and T‐ zones, as well as at the subcapsular sinus in LNs, the location where LTi cells reside in murine spleen and LNs. Furthermore, like murine LTi cells, these cells expressed high levels of IL‐22 and upregulated IL‐22 expression upon IL‐23 stimulation. Importantly, these cells were not an NK cell subset since they showed no expression of IFN‐γ and perforin. Interestingly, a subset of the CD117<SUP>+</SUP>CD3<SUP>−</SUP>CD56<SUP>−</SUP>OX40L<SUP>+</SUP> population expressed NKp46, again similar to recent findings in mice. Finally, these cells supported memory CD4<SUP>+</SUP> T‐cell survival in an OX40L‐dependent manner. Combined, these data indicate that the CD117<SUP>+</SUP>CD3<SUP>−</SUP>CD56<SUP>−</SUP>OX40L<SUP>+</SUP> cells in human secondary lymphoid tissues are comparable in phenotype, location and function to the LTi cells that persist within adult murine secondary lymphoid tissues.</P>