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Reduced Ceramides Are Associated with Acute Rejection in Liver Transplant Patients and Skin Graft and Hepatocyte Transplant Mice, Reducing Tolerogenic Dendritic Cells

  • Hyun Ju Yoo;Yeogyeong Yi;Yoorha Kang;Su Jung Kim;Young-In Yoon;Phuc Huu Tran;Taewook Kang;Min Kyung Kim;Jaeseok Han;Eunyoung Tak;Chul-Soo Ahn;Gi-Won Song;Gil-Chun Park;Sung-Gyu Lee;Jae-Joong Kim;Dong-Hwan Jung;Shin Hwang;Nayoung Kim
    • Molecules and Cells
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    • 제46권11호
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    • pp.688-699
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    • 2023
  • We set up this study to understand the underlying mechanisms of reduced ceramides on immune cells in acute rejection (AR). The concentrations of ceramides and sphingomyelins were measured in the sera from hepatic transplant patients, skin graft mice and hepatocyte transplant mice by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Serum concentrations of C24 ceramide, C24:1 ceramide, C16:0 sphingomyelin, and C18:1 sphingomyelin were lower in liver transplantation (LT) recipients with than without AR. Comparisons with the results of LT patients with infection and cardiac transplant patients with cardiac allograft vasculopathy in humans and in mouse skin graft and hepatocyte transplant models suggested that the reduced C24 and C24:1 ceramides were specifically involved in AR. A ceramide synthase inhibitor, fumonisin B1 exacerbated allogeneic immune responses in vitro and in vivo, and reduced tolerogenic dendritic cells (tDCs), while increased P3-like plasmacytoid DCs (pDCs) in the draining lymph nodes from allogeneic skin graft mice. The results of mixed lymphocyte reactions with ceranib-2, an inhibitor of ceramidase, and C24 ceramide also support that increasing ceramide concentrations could benefit transplant recipients with AR. The results suggest increasing ceramides as novel therapeutic target for AR, where reduced ceramides were associated with the changes in DC subsets, in particular tDCs.

Soluble Prokaryotic Expression and Purification of Bioactive Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand

  • Do, Bich Hang;Nguyen, Minh Tan;Song, Jung-A;Park, Sangsu;Yoo, Jiwon;Jang, Jaepyeong;Lee, Sunju;So, Seoungjun;Yoon, Yejin;Kim, Inki;Lee, Kyungjin;Jang, Yeon Jin;Choe, Han
    • Journal of Microbiology and Biotechnology
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    • 제27권12호
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    • pp.2156-2164
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    • 2017
  • Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b'a' domain of protein disulfide isomerase (PDIb'a'), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was $0.4EU/{\mu}g$, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an $EC_{50}$ and Hill coefficient of $0.6{{\pm}}0.03nM$ and $2.41{\pm}0.15$, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

The First Living-Donor Lobar Lung Transplantation in Korea: a Case Report

  • Choi, Sehoon;Park, Seung-Il;Lee, Geun Dong;Kim, Hyeong Ryul;Kim, Dong Kwan;Jung, Sung-Ho;Yun, Tae-Jin;Kim, In Ok;Choi, Dae-Kee;Choi, In-Cheol;Song, Jong-Min;Hong, Sang-Bum;Shim, Tae Sun;Jo, Kyung-Wook;Lee, Sang-Oh;Do, Kyung-Hyun;Chae, Eun Jin
    • Journal of Korean Medical Science
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    • 제33권43호
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    • pp.282.1-282.6
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    • 2018
  • Lung transplantation is the only treatment for end-stage lung disease, but the problem of donor shortage is unresolved issue. Herein, we report the first case of living-donor lobar lung transplantation (LDLLT) in Korea. A 19-year-old woman patient with idiopathic pulmonary artery hypertension received her father's right lower lobe and her mother's left lower lobe after pneumonectomy of both lungs in 2017. The patient has recovered well and is enjoying normal social activity. We think that LDLLT could be an alternative approach to deceased donor lung transplantation to overcome the shortage of lung donors.

Iris Nertschinskia Ethanol Extract Differentially Induces Cytotoxicity in Human Breast Cancer Cells Depending on AKT1/2 Activity

  • Shin, Jae-Sik;Maeng, Hyung-Gun;Hong, Seung-Woo;Moon, Jai-Hee;Kim, Jin-Sun;Suh, Young-Ah;Kim, Eun-Sung;Lee, Young-Min;Kim, Ye-Seul;Choi, Eun-Kyung;Kim, Inki;Lee, Sok-Young;Cho, Dong-Hyung;Hong, Nam-Joo;Kim, Tae-Won;Jin, Dong-Hoon;Lee, Wang Jae
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권12호
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    • pp.6511-6516
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    • 2012
  • Recently, we reported that an ethanol extract of Iris nertschinskia induces p53-dependent apoptosis in the MCF7 human breast cancer cell line. However, the detailed mechanisms were not fully explored. Here, we demonstrate another aspect of the activity of I. nertschinskia in breast cancer cells. We compared the response to an ethanol extract of I. nertschinskia in two different human breast cancer cell lines, Hs578Tand MDA-MB231, respectively with relatively low and high AKT1/2 activity by trypan blue exclusion assay and FACS analysis. Knockdown of endogenous AKT1 or AKT2 in breast cancer cells by RNA interference determined the sensitivity to I. nertschinskia ethanol extract compared to control cells. The I. nertschinskia ethanol extract induced cell death in a manner that depended on the level of phosphorylated AKT1/2 protein and was associated with a significant increase in the sub-G1 cell population, indicative of apoptosis. Our results indicate that an ethanol extract of I. nertschinskia differentially induces cell death in breast cancer cells depending on their level of phosphorylated AKT1/2.

Characterizing Organelles in Live Stem Cells Using Label-Free Optical Diffraction Tomography

  • Kim, Youngkyu;Kim, Tae-Keun;Shin, Yeonhee;Tak, Eunyoung;Song, Gi-Won;Oh, Yeon-Mok;Kim, Jun Ki;Pack, Chan-Gi
    • Molecules and Cells
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    • 제44권11호
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    • pp.851-860
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    • 2021
  • Label-free optical diffraction tomography (ODT), an imaging technology that does not require fluorescent labeling or other pre-processing, can overcome the limitations of conventional cell imaging technologies, such as fluorescence and electron microscopy. In this study, we used ODT to characterize the cellular organelles of three different stem cells-namely, human liver derived stem cell, human umbilical cord matrix derived mesenchymal stem cell, and human induced pluripotent stem cell-based on their refractive index and volume of organelles. The physical property of each stem cell was compared with that of fibroblast. Based on our findings, the characteristic physical properties of specific stem cells can be quantitatively distinguished based on their refractive index and volume of cellular organelles. Altogether, the method employed herein could aid in the distinction of living stem cells from normal cells without the use of fluorescence or specific biomarkers.