Enhanced antitumor immunotherapeutic effect of B-cell-based vaccine transduced with modified adenoviral vector containing type 35 fiber structures

Kim, E-K,Seo, H-S,Chae, M-J,Jeon, I-S,Song, B-Y,Park, Y-J,Ahn, H M,Yun, C-O,Kang, C-Y Macmillan Publishers Limited 2014 Gene therapy Vol.21 No.1

For successful clinical tumor immunotherapy outcomes, strong immune responses against tumor antigens must be generated. Cell-based vaccines compromise one strategy with which to induce appropriate strong immune responses. Previously, we established a natural killer T-cell (NKT) ligand-loaded, adenoviral vector-transduced B-cell-based anticancer cellular vaccine. To enhance tumor antigen delivery to B cells, we established a modified adenoviral vector (Ad-k35) that encoded a truncated form of the breast cancer antigen Her2/neu (Ad-k35HM) in which fiber structure was substituted with adenovirus serotype 35. We observed increased tumor antigen expression with Ad-k35HM in both human and murine B cells. In addition, an Ad-k35HM-transduced B-cell vaccine elicited strong antigen-specific cellular and humoral immune responses that were further enhanced with the additional loading of soluble NKT ligand KBC009. An Ad-k35HM-transduced, KBC009-loaded B-cell vaccine efficiently suppressed the in vivo growth of established tumors in a mouse model. Moreover, the vaccine elicited human leukocyte antigen (HLA)-A2 epitope-specific cytotoxic T-cell responses in B6.Cg (CB)-Tg (HLA-A/H2-D) 2Enge/Jat mice. These findings indicated that the Ad-k35 could be appropriate for the preclinical and clinical development of B-cell-based anticancer immunotherapies.

Inhibitory effect of obovatol on nitric oxide production and activation of NF-κB/MAP kinases in lipopolysaccharide-treated RAW 264.7cells

Choi, M.S.,Lee, S.H.,Cho, H.S.,Kim, Y.,Yun, Y.P.,Jung, H.Y.,Jung, J.K.,Lee, B.C.,Pyo, H.B.,Hong, J.T. North-Holland ; Elsevier Science Ltd 2007 european journal of pharmacology Vol.556 No.1

The components of Magnolia obovata are known to have many pharmacological activities. In this study, we investigated the effects of obovatol, a neolignan compound isolated from the leaves of M. obovata, on nitric oxide (NO) production and NF-κB activity in lipopolysaccharide (LPS)-activated RAW 264.7 cells. The results show that obovatol (1-5 μM) significantly inhibited LPS-induced NO production in a concentration-dependent manner (IC<SUB>50</SUB>: 0.91 μM). Consistent with the inhibitory effect on NO production, obovatol inhibits the expression of inducible nitric oxide synthase and cyclooxygenase-2 expression. Furthermore, obovatol suppressed NF-κB (p50 and p65) translocation to the nucleus as well as IκB release resulting in the inhibition of the DNA binding activity of the NF-κB. Obovatol also inhibited c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) signal, which are the most significantly involved signal in NO production and NF-κB activation. When the cells were treated with the combination of obovatol with U0126 (an ERK inhibitor) or SP600125 (a JNK inhibitor) as well as with SC-514 (an IKK2 inhibitor), much more inhibition of NO production was observed than that by obovatol alone. The present results suggest that obovatol has an inhibitory effect on NO production through the inhibition of NF-κB/MAPK activity, and thus can be used as an anti-inflammatory agent.

Efficacy of combining ING4 and TRAIL genes in cancer-targeting gene virotherapy strategy: first evidence in preclinical hepatocellular carcinoma

Galal El-Shemi, A,Mohammed Ashshi, A,Oh, E,Jung, B-K,Basalamah, M,Alsaegh, A,Yun, C-O Nature Publishing Group 2018 Gene Therapy Vol.25 No.1

<P>Current treatments of hepatocellular carcinoma (HCC) are ineffective and unsatisfactory in many aspects. Cancer-targeting gene virotherapy using oncolytic adenoviruses (OAds) armed with anticancer genes has shown efficacy and safety in clinical trials. Nowadays, both inhibitor of growth 4 (ING4), as a multimodal tumor suppressor gene, and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), as a potent apoptosis-inducing gene, are experiencing a renaissance in cancer gene therapy. Herein we investigated the antitumor activity and safety of mono- and combined therapy with OAds armed with ING4 (Ad-ΔB/ING4) and TRAIL (Ad-ΔB/TRAIL) gene, respectively, on preclinical models of human HCC. OAd-mediated expression of ING4 or TRAIL transgene was confirmed. Ad-ΔB/TRAIL and/or Ad-ΔB/ING4 exhibited potent killing effect on human HCC cells (HuH7 and Hep3B) but not on normal liver cells. Most importantly, systemic therapy with Ad-ΔB/ING4 plus Ad-ΔB/TRAIL elicited more eradicative effect on an orthotopic mouse model of human HCC than their monotherapy, without causing obvious overlapping toxicity. Mechanistically, Ad-ΔB/ING4 and Ad-ΔB/TRAIL were remarkably cooperated to induce antitumor apoptosis and immune response, and to repress tumor angiogenesis. This is the first study showing that concomitant therapy with Ad-ΔB/ING4 and Ad-ΔB/TRAIL may provide a potential strategy for HCC therapy and merits further investigations to realize its possible clinical translation.</P>

ERK-dependent IL-6 autocrine signaling mediates adaptive resistance to pan-PI3K inhibitor BKM120 in head and neck squamous cell carcinoma

Yun, M R,Choi, H M,Kang, H N,Lee, Yw,Joo, H-S,Kim, D H,Kim, H R,Hong, M H,Yoon, S O,Cho, B C Nature Publishing Group 2018 Oncogene Vol. No.

Hyperactivation of phosphatidylinositol 3-kinase (PI3K) pathway occurs frequently in head and neck squamous cell carcinoma (HNSCC). However, clinical outcomes of targeting the PI3K pathway have been underwhelming. In present study, we investigated the resistant mechanisms and potential combination therapeutic strategy to overcome adaptive resistance to PI3K inhibitor in HNSCC. Treatment of NVP-BKM120, a pan-PI3K inhibitor, led to upregulation of interleukin-6 (IL-6) and subsequent activation of either extracellular signal-regulated kinase (ERK) or signal transducers and activators of transcription 3 (STAT3), causing modest antitumor effects on the growth of HNSCC cells. Blockade of autocrine IL-6 signaling with siRNA or neutralizing antibody for IL-6 receptor (IL-6R) completely abolished NVP-BKM120-induced activation of ERK and STAT3 as well as expression of c-Myc oncogene, which resulted in enhanced sensitivity to NVP-BKM120. Moreover, when compared with a pharmacologic inhibitor or silencing of STAT3, trametinib, a MEK inhibitor, in combination with NVP-BKM120 yielded more potent anti-proliferative effects by inhibiting S phase transition, arresting cells at G0/G1 phase, and downregulating IL-6 and c-Myc expression. Furthermore, as compared with either agent alone, combination of NVP-BKM120 with trametinib or tocilizumab, a humanized anti-IL-6R antibody, significantly suppressed tumor growth in NVP-BKM120-resistant patient-derived tumor xenograft (PDTX) models, which was also confirmed in PDTX-derived cell lines. Collectively, these results suggested that IL-6/ERK signaling is closely involved in adaptive resistance of NVP-BKM120 in HNSCC cells, providing a rationale for a novel combination therapy to overcome resistance to PI3K inhibitors.

Performance test of new near-ambient-pressure XPS at Korean Basic Science Institute and its application to CO oxidation study on Pt₃Ti polycrystalline surface

Jeong, C.,Yun, H.,Lee, H.,Muller, S.,Lee, J.,Mun, B.S. Elsevier 2016 CURRENT APPLIED PHYSICS Vol.16 No.1

<P>A performance test of near-ambient pressure X-ray photoelectron spectroscopy (NAP-XPS) at Korean Basic Science Institute (KBSI) in Daejeon, Korea is carried out. The NAP-XPS at KBSI is equipped with SPECS PHOIBOS 150 analyzer and 2-dimensional-delay line detector, which provides improved detection efficiency with 1-dimensional chemical imaging mode. The in-situ gas-pressure cell is designed to operate at pressure up to 25 mbar with differential pumping scheme. A micro-focused AlKa X-ray source is utilized as photon excitation source. The NAP-XPS is composed of two separate chambers, e.g. a main preparation chamber and a measurement chamber, which hold a low energy electron diffraction, a high pressure cell, an ion sputter gun, a multi-cell E-beam evaporators, and a four-axis manipulator with heating and cooling capability. As a test run of NAP-XPS, a CO oxidation experiment is carried out on Pt3Ti polycrystalline surface. During the active phase of CO oxidation, a clear formation of Ti oxide is found on surface, revealing the intriguing role of surface metal oxide in surface chemical reaction. (C) 2015 Elsevier B.V. All rights reserved.</P>

Variable number of tandem repeats of 9 Plasmodium vivax genes among Southeast Asian isolates

Wang, B.,Nyunt, M.H.,Yun, S.G.,Lu, F.,Cheng, Y.,Han, J.H.,Ha, K.S.,Park, W.S.,Hong, S.H.,Lim, C.S.,Cao, J.,Sattabongkot, J.,Kyaw, M.P.,Cui, L.,Han, E.T. Verlag fur Recht und Gesellschaft 2017 Acta tropica Vol.170 No.-

<P>The variable number of tandem repeats (VNTRs) provides valuable information about both the functional and evolutionary aspects of genetic diversity. Comparative analysis of 3 Plasmodium falciparum genomes has shown that more than 9% of its open reading frames (ORFs) harbor VNTRs. Although microsatellites and VNTR genes of P. vivax were reported, the VNTR polymorphism of genes has not been examined widely. In this study, 230 P. vivax genes were analyzed for VNTRs by SERV, and 33 kinds of TR deletions or insertions from 29P. vivax genes (12.6%) were found. Of these, 9 VNTR fragments from 8 P. vivax genes were used for PCR amplification and sequence analysis to examine the genetic diversity among 134 isolates from four Southeast Asian countries (China, Republic of Korea, Thailand, and Myanmar) with different malaria endemicity. We confirmed the existence of extensive polymorphism of VNTR fragments in field isolates. This detection provides several suitable markers for analysis of the molecular epidemiology of P. vivax field isolates. (C) 2017 Elsevier B.V. All rights reserved.</P>

Gut-Specific Delivery of T-Helper 17 Cells Reduces Obesity and Insulin Resistance in Mice

Hong, C.P.,Park, A.,Yang, B.G.,Yun, C.H.,Kwak, M.J.,Lee, G.W.,Kim, J.H.,Jang, M.S.,Lee, E.J.,Jeun, E.J.,You, G.,Kim, K.S.,Choi, Y.,Park, J.H.,Hwang, D.,Im, S.H.,Kim, J.F.,Kim, Y.K.,Seoh, J.Y.,Surh, C. Elsevier North Holland [etc.] 2017 Gastroenterology Vol.152 No.8

<P>BACKGROUND & AIMS: Obesity and metabolic syndrome have been associated with alterations to the intestinal microbiota. However, few studies examined the effects of obesity on the intestinal immune system. We investigated changes in subsets of intestinal CD4(+) T-helper (T-H) cells with obesity and the effects of gut-tropic T(H)17 cells in mice on a high-fat diet (HFD). METHODS: We isolated immune cells from small intestine and adipose tissue of C57BL/6 mice fed a normal chow diet or a HFD for 10 weeks and analyzed the cells by flow cytometry. Mice fed a vitamin A-deficient HFD were compared with mice fed a vitamin A-sufficient HFD. Obese RAG1-deficient mice were given injections of only regulatory T cells or a combination of regulatory T cells and T(H)17 cells (wild type or deficient in integrin beta 7 subunit or interleukin 17 [IL17]). Mice were examined for weight gain, fat mass, fatty liver, glucose tolerance, and insulin resistance. Fecal samples were collected before and after T cell transfer and analyzed for microbiota composition by metagenomic DNA sequencing and quantitative polymerase chain reaction. RESULTS: Mice placed on a HFD became obese, which affected the distribution of small intestinal CD4(+) T-H cells. Intestinal tissues from obese mice had significant reductions in the proportion of T(H)17 cells but increased proportion of T(H)1 cells, compared with intestinal tissues from nonobese mice. Depletion of vitamin A in obese mice further reduced the proportion of T(H)17 cells in small intestine; this reduction correlated with more weight gain and worsening of glucose intolerance and insulin resistance. Adoptive transfer of in vitro-differentiated gut-tropic T(H)17 cells to obese mice reduced these metabolic defects, which required the integrin beta 7 subunit and IL17. Delivery of T(H)17 cells to intestines of mice led to expansion of commensal microbes associated with leanness. CONCLUSIONS: In mice, intestinal T(H)17 cells contribute to development of a microbiota that maintains metabolic homeostasis, via IL17. Gut-homing T(H)17 cells might be used to reduce metabolic disorders in obese individuals.</P>

Phosphorylation of CHIP at Ser20 by Cdk5 promotes tAIF-mediated neuronal death

Kim, C,Yun, N,Lee, J,Youdim, M B H,Ju, C,Kim, W-K,Han, P-L,Oh, Y J Macmillan Publishers Limited 2016 CELL DEATH AND DIFFERENTIATION Vol.23 No.2

<P>Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/ threonine kinase and its dysregulation is implicated in neurodegenerative diseases. Likewise, C-terminus of Hsc70-interacting protein (CHIP) is linked to neurological disorders, serving as an E3 ubiquitin ligase for targeting damaged or toxic proteins for proteasomal degradation. Here, we demonstrate that CHIP is a novel substrate for Cdk5. Cdk5 phosphorylates CHIP at Ser20 via direct binding to a highly charged domain of CHIP. Co-immunoprecipitation and ubiquitination assays reveal that Cdk5-mediated phosphorylation disrupts the interaction between CHIP and truncated apoptosis-inducing factor (tAIF) without affecting CHIP's E3 ligase activity, resulting in the inhibition of CHIP-mediated degradation of tAIF. Lentiviral transduction assay shows that knockdown of Cdk5 or overexpression of CHIPS20A, but not CHIPWT, attenuates tAIF-mediated neuronal cell death induced by hydrogen peroxide. Thus, we conclude that Cdk5-mediated phosphorylation of CHIP negatively regulates its neuroprotective function, thereby contributing to neuronal cell death progression following neurotoxic stimuli.</P>

Performance characteristics of a 200-kW organic Rankine cycle system in a steel processing plant

Sung, T.,Yun, E.,Kim, H.D.,Yoon, S.Y.,Choi, B.S.,Kim, K.,Kim, J.,Jung, Y.B.,Kim, K.C. Applied Science Publishers 2016 APPLIED ENERGY Vol.183 No.-

<P>The main objective of this research is to design and build a 200-kW ORC system with reduced size that could be installed in a steel processing plant where space is limited. The real-time operating characteristics of the ORC system are demonstrated with actual flue gases. First, an ORC system with R245fa refrigerant was developed for a heat source temperature of 140 degrees C. The evaporation and condensation pressures were 2,090 kPa and 220 kPa, respectively. The net power output was 235.7 kW with a thermal efficiency of 12.9%. Using an electric heat source, the design point performance of the system is experimentally demonstrated and shows a net power output of 177.4 kW with thermal efficiency of 9.6%. The turbine isentropic efficiency and generator efficiency were 68.1% and 98.5% at a rotational speed of 14,000 rpm. Next, the ORC system was implemented by designing a dedicated heat transfer loop for a steel processing plant using data measured from a chimney. The experimental net power output is 105.8 kW with a thermal efficiency of 8.6% when the plant is operated at the highest work load. The fluctuation of the flue gas temperature is successfully suppressed with a thermal storage tank installed in the heat transfer loop. A partial-load analysis was conducted and showed that the system has the highest performance with more than 165 kW of net power output. Economic analysis of such system showed that the right sized ORC system with always working parent plant had good economics with a payback period of 9 years. (C) 2016 Elsevier Ltd. All rights reserved.</P>

Measurement of Viable Cell Number in Mixed Culture Based on Microbial Respiration Rate

Veljkoic, V.B.,Kwon, Yun Joong,Engler, C.R. 한국산업미생물학회 1992 한국미생물·생명공학회지 Vol.20 No.6

혼합배양중의 각 미생물의 생균수 측정은 순수배양보다 훨씬 복잡하다. 특히 두 균주의 크기가 비슷한 경우에는 사용할 수 있는 방법이 더 제한된다. 본 연구에서는 두 균의 크기가 비슷한 경우에도 적용될 수 있는 간단한 생균수 측정방법을 개발하였다. 미생물 배양액의 산소흡수속도(OUR)는 세포수에 비례하며 이때의 비례상수인 최대 비산소흡수속도(maximum specific OUR)를 알고 있으면 배양액의 OUR을 측정함으로써 간접적으로 생균수를 구할 수 있게 된다. 혼합배양의 경우 산소흡수속도는 각 미생물의 호흡속도의 합이 되며, 각 미생물의 호흡속도가 서로 다르고 또한 온도의존성이 다르다면 호흡 속도의 측정을 이용하여 각 생균수를 간접적으로 측정할 수 있다. 산소흡수속도는 시가에 따른 용존산소 농도의 변화를 DO probe를 이용하여 측정하여 구했으며, 최대 비산소흡수속도는 plate count에 의한 생균수와 OUR의 직선관계의 기울기에서 구했다. C. lusitaniae의 최대 비산소흡수속도는 20과 30℃에서 각각 1.36×exp(-19)과 3.90×exp(-9), P. tannophilus는 20과 30 ℃에서 각각 0.59×exp(-9)와 1.86×exp(-9)[(%/s)/(cells)/㎖]]이었다. 이들 값을 이용하여 계산한 혼하배양액 중의 두 효모의 생균수와 서로 다른 배지를 이용하여 plate count로 측정한 생균수와의 관계는 상관계수 0.98로서 비교적 잘 일치되었다. A simple method to determine viable cell numbers of each species in mixed culture was developed. The oxygen uptake rate (OUR) equals to the product of the specific OUR and the size of the microbial population. In a mixed culture, the OUR is a result of the respiration activities of each sub-population. The OUR was determined from the slope of the linear relationship between time and the decrease of dissolved oxygen concentration when aeration was stopped. The specific OUR was calculated from the slope of the viable cell number versus OUR curve. These values for C. lusitaniae at 20 and 30℃ were 1.36×exp(-19) and 3.90×exp(-9) and those for P. tannophilus at 20 and 30℃ were 0.59×exp(-19) and 1.86×exp(-9) [(%/s)/(cells/㎖)], respectively. Using these values, viable cell numbers were calculated after the OURs of mixed culture at two temperatures were measured. A good agreement between the viable cell numbers determined by this method and by plate count was obtained.