• Title/Summary/Keyword: Vaccine production

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High Level Production of Glycoprotein H of HSV-1 (F) Using HcNPV Vector System

  • Kang, Hyun;Cha, Soung-Chul;Han, You-Jin;Park, In-Ho;Lee, Min-Jung;Byun, Si-Myung;Lee, Hyung-Hoan
    • BMB Reports
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    • v.33 no.6
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    • pp.483-492
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    • 2000
  • The Herpes simplex virus type 1 (HSV-1) strain F glycoprotein H (gH) gene in the pHLB-4 plasmid was recombinated into a baculovirus expression vector (lacZ-HcNPV) to construct a recombinant virus GH-HcNPV expressing gH. The sequences of gH and its expression were analyzed. The gH gene was located in the 6.41 kb BglII fragment. The open reading frame (ORF) of the gH gene was 2,517 bp and codes 838 amino acid residues. Insect cells infected with this recombinant virus synthesized a high level of the matured and gX-gH fusion protein with approximately 112 kDa. The fusion gH protein was localized on the membrane of the insect cells as seen by using immunofluorescence assay and accumulated in the cultured media by the SDS-PAGE and immunoprecipitation assays. The amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. Antibodies raised in mice to this recombinant protein recognized viral gH and neutralized the infectivity of HSV-1 in vitro. These results demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV; accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

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Bee Venom (Apis Mellifera) an Effective Potential Alternative to Gentamicin for Specific Bacteria Strains - Bee Venom an Effective Potential for Bacteria-

  • Zolfagharian, Hossein;Mohajeri, Mohammad;Babaie, Mahdi
    • Journal of Pharmacopuncture
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    • v.19 no.3
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    • pp.225-230
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    • 2016
  • Objectives: Mellitine, a major component of bee venom (BV, Apis mellifera), is more active against gram positive than gram negative bacteria. Moreover, BV has been reported to have multiple effects, including antibacterial, antivirus, and anti-inflammation effects, in various types of cells. In addition, wasp venom has been reported to have antibacterial properties. The aim of this study was to evaluate the antibacterial activity of BV against selected gram positive and gram negative bacterial strains of medical importance. Methods: This investigation was set up to evaluate the antibacterial activity of BV against six grams positive and gram negative bacteria, including Staphylococcus aureus (S. aureus), Salmonella typhimurium, Escherichia coli (E. coli) O157:H7, Pseudomonas aeruginosa, Burkholderia mallei and Burkholderia pseudomallei. Three concentrations of crude BV and standard antibiotic (gentamicin) disks as positive controls were tested by using the disc diffusion method. Results: BV was found to have a significant antibacterial effect against E. coli, S. aureus, and Salmonella typhyimurium in all three concentrations tested. However, BV had no noticeable effect on other tested bacteria for any of the three doses tested. Conclusion: The results of the current study indicate that BV inhibits the growth and survival of bacterial strains and that BV can be used as a complementary antimicrobial agent against pathogenic bacteria. BV lacked the effective proteins necessary for it to exhibit antibacterial activity for some specific strains while being very effective against other specific strains. Thus, one may conclude, that Apis mellifera venom may have a specific mechanism that allows it to have an antibacterial effect on certain susceptible bacteria, but that mechanism is not well understood.

Characteristics of Transmissible CTX-M- and CMY-Type β-Lactamase-Producing Escherichia coli Isolates Collected from Pig and Chicken Farms in South Korea

  • Shin, Seung Won;Jung, Myunghwan;Won, Ho Geun;Belaynehe, Kuastros Mekonnen;Yoon, In Joong;Yoo, Han Sang
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1716-1723
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    • 2017
  • The rapid dissemination of extended-spectrum ${\beta}$-lactamase (ESBL)-producing Escherichia coli has significantly contributed to public health hazard globally. A total of 281 E. coli strains recovered from pigs and chickens between 2009 and 2015 in South Korea were analyzed for ESBL production. ESBL phenotypes were recognized in 14 E. coli isolates; ten and three ESBL-producing isolates carried only $bla_{CTX-M}$ and $bla_{CMY}$ genes, respectively, and one isolate harbored both genes. The predominant CTX-M and CMY types were CTX-M-15 (n = 8) and CMY-2 (n = 3). We also detected ESBL-producing isolates harboring $bla_{CTX-M-65}$, $bla_{CTX-M-14}$, $bla_{CMY-6}$, $bla_{DHA-1}$, and $bla_{TEM-1}$ genes. All ESBL-producing isolates showed resistance to the extent of the fourth generation cephalosporins, along with multidrug resistance. CTX-M-15-producing isolates showed higher MIC values than CTX-M-14- and CTX-M-65-producing isolates. The $bla_{CTX-M}$ and $bla_{CMY}$ genes have the potential to be transferable. The spreading of $bla_{CMY}$ and $bla_{CTX-M}$ genes was arbitrated mainly via Frep and IncI1 plasmids. Our isolates showed clonal diversity in PFGE analysis. This is the first report of E. coli isolates carrying $bla_{CMY-6}$ in chicken from South Korea. The emergence of CMY-6 ESBLs in a population of poultry suggests that extensive screening with long-term surveillance is necessary to prevent the dissemination of ESBL from chicken to human.

Immunoadjuvant Activity of Korean Mistletoe Lectin B-chain (한국산 겨우살이 Lectin B-chain의 면역증강 효과)

  • Her, Sun-Mi;An, Hyo-Sun;Kim, Kyu-Dae;Kim, Young-Hoon;Kim, In-Bo;Yoon, Taek-Joon;Kim, Jong-Bae
    • Korean Journal of Pharmacognosy
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    • v.42 no.3
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    • pp.246-252
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    • 2011
  • Korean mistletoe Lectin (KML-C) is composed of A and B sub-chain. B chain binds to carbohydrates on cell surface and A chain hinders translation and induces an apoptosis as a RIP (ribosome inactivating protein). KML-C has very strong biological activities, it has seriously limits to use as a cancer therapy or adjuvant because of its toxicity to normal cells. This study is therefore conducted to see if B chain of KML-C might have immunological activity, especially adjuvant activities with less toxicity. We isolated B chain from KML-C using the lactose affinity chromatography, and examined their immunoadjuvant activity. The isolated B-chain did not show any cytotoxicity against tumor cell, RAW264.7, and P388D1 while KML-C had a very strong toxicity. This non-toxic effect was observed also by in-vivo study. Both humoral and cellular immunities were observed ; the antibody titer was increased when the mice were immunized with B-chain used as adjuvant like Freund's adjuvant, indicating that B chain of mistletoe lectin alone might be used for adjuvant; it also increased DTH in cellular immunity. These results suggest that B-chain of KML-C might be used for adjuvant used for the production of antibody or vaccine with less toxicity.

Current Status and Future Directions of Fish Vaccines in Korea (국내 수산용 백신 개발 현황 및 발전 방향)

  • Han, Hyun-Ja;Kim, Soo-Jin;Kim, Tae-Ho;Kim, Myoung-Sug;Cho, Mi-Young;Choi, Hye Sung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.4
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    • pp.369-376
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    • 2021
  • In Korea, fish is a major food source, and sustainable production of fish is critical for the aquaculture industry. Recently, infectious diseases have become an unavoidable problem in aquaculture and have caused serious economic losses. Therefore, there is an urgent need to overcome this challenge to increase productivity in aquaculture. Vaccination is the most effective and long-term measure for improving the management of a fish farm. Sales of fish vaccines in Korea have been steadily increasing, with a confirmed increase from 600 million won in 2007 to 3.4 billion won in 2018. As of 2021, 30 licensed fish vaccines, including both single and combination vaccines containing more than two inactivated pathogens, have been approved in Korea for 10 fish pathogens. Twenty-eight of these are used in olive flounder Paralichthys olivaceus; however, only two vaccines are used in starry flounder Platichthys stellatus, red seabream Ragrus major and rock bream Oplegnathus fasciatus. Therefore, vaccine development is required not only for the olive flounder but also for other cultured fish species. The development of effective vaccines and vaccination programs for fish is a continuous requirement for a sustainable aquaculture industry.

Biochemical, Hematological Effects and Complications of Pseudosynanceia Melanostigma Envenoming

  • Babaie, Mahdi;Zolfagharian, Hossein;Zolfaghari, Mohammad;Jamili, Shahla
    • Journal of Pharmacopuncture
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    • v.22 no.3
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    • pp.140-146
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    • 2019
  • Objectives: Venomous fishes have different pharmacological effects and are useful. Among the venomous fish, stonefishes; especially Pseudosynanceia melanostigma has various pharmacological effects on the nervous, muscular and cardiovascular system of humans. In this study, toxicological characteristics, some blood effects, pharmacological and enzymatic properties of Pseudosynanceia melanostigma venom was investigated. Methods: Crude venom purified by using gel filtration chromatography and the molecular weights of the venom and its fractions were estimated. The approximate LD values of this venom were determinedand the effects of LD50 dose on the blood of rabbits were studied. Hemolytic and Hemorrhagic activity of the venom sample was determined. In this case coagulation tests were performed. Results: The LD50 of the Pseudosynanceia melanostigma crude venom was also determined to be $194.54{\mu}g/mouse$. The effect of two doses of LD50 showed a non-significant differences decrease in RBCs and MCV. In other cases, the results showed significant differences in WBC, Plt, Hb, MCH, MCHC and HCT; also it's showed a significant decrease. WBC count showed a significant increase with two doses of LD50 groups. The prothrombin time and partial prothrombin time were increased after venom treatment. As well as bleeding and clotting time were increased. According to the results, a minimum dose for Haemorrhagic effect $40{\mu}g$ was obtained. Conclusion: Venom of Pseudosynanceia melanostigma has inhibitory effect on platelet aggregation that can be used to design and develop of anticoagulant drugs.

Bibliometric analysis on the evolution of knowledge structure of African swine fever

  • Oh, Jee-Sun;Cho, Ho-Seong;Oh, Yeonsu
    • Korean Journal of Veterinary Service
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    • v.44 no.4
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    • pp.257-270
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    • 2021
  • Since African swine fever (ASF) spread to East Asia, a fatal crisis has occurred in the global pig industry, because Asia is dominant in pig production. Although some studies conducted bibliometric analysis on ASF, few studies compared research networks, and identified subthemes by major keywords. To fill this gap, this study identified the knowledge structure network of the research, its influence, and core research themes by utilizing the bibliometric analysis of 337 ASF-related journal articles over 50 years from 1970 to 2020 on the Web of Science. The result indicated that papers are mainly published in the fields of veterinary science, virology, microbiology, infectious disease and applied microbiology, and in particular, the fields of veterinary science and virology showed unrivaled weights as they account for 73.40%. With regard to cooperative relationships, European countries such as the UK, Germany, Italy, and Denmark, centered on Spain, are actively contributing to the ASF research. China, France, Thailand, Japan, Vietnam, and South Korea are leading research cooperation, centering on the United States. In the early stage of the studies, major keywords appeared to be related to outbreaks, quarantine and diagnosis, and in the middle stage, the keywords were expanded to a wide range of pig diseases. Recently, the keywords are becoming more diverse towards antibodies, cross-border transmission and disease monitoring. Based on data on major keywords related to ASF, this study proposed discussions and implications for activating ASF research including genotype, protein, vaccine, diagnosis, defense against infection and epidemiological investigation.

Deinococcus radiodurans R1 Lysate Induces Tolerogenic Maturation in Lipopolysaccharide-Stimulated Dendritic Cells and Protects Dextran Sulfate Sodium-Induced Colitis in Mice

  • Song, Ha-Yeon;Han, Jeong Moo;Kim, Woo Sik;Lee, Ji Hee;Park, Woo Yong;Byun, Eui-Baek;Byun, Eui-Hong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.835-843
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    • 2022
  • Deinococcus radiodurans is an extremophilic bacterium that can thrive in harsh environments. This property can be attributed to its unique metabolites that possess strong antioxidants and other pharmacological properties. To determine the potential of D. radiodurans R1 lysate (DeinoLys) as a pharmacological candidate for inflammatory bowel disease (IBD), we investigated the antiinflammatory activity of DeinoLys in bone marrow-derived dendritic cells (BMDCs) and a colitis mice model. Lipopolysaccharide (LPS)-stimulated BMDCs treated with DeinoLys exhibited alterations in their phenotypic and functional properties by changing into tolerogenic DCs, including strongly inhibited proinflammatory cytokines (TNF-α and IL-12p70) and surface molecule expression and activated DC-induced T cell proliferation/activation with high IL-10 production. These phenotypic and functional changes in BMDCs induced by DeinoLys in the presence of LPS were abrogated by IL-10 neutralization. Furthermore, oral administration of DeinoLys significantly reduced clinical symptoms against dextran sulfate sodium-induced colitis, including body weight loss, disease activity index, histological severity in colon tissue, and lower myeloperoxidase level in mice. Our results establish DeinoLys as a potential anti-inflammatory candidate for IBD therapy.

Induction of Autophagy by Rosa acicularis Leaves Extracts in RAW264.7 Cells (인가목(Rosa acicularis Lindl.) 잎 추출물의 대식세포에서 자가포식 유도활성)

  • Jeong Won Choi;Jin Boo Jeong
    • Korean Journal of Plant Resources
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    • v.36 no.4
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    • pp.257-263
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    • 2023
  • Autophagy contributes to enhancing the immune system (innate and adaptive immune system) against foreign pathogens. Autophagy of macrophages is used as a major indicator for developing vaccine adjuvants to increase the adaptive immune response. In this study, water extracts from Rosa acicularis leaves (RAL) increased the production of immunostimulatory mediators and phagocytic activity in RAW264.7 cells. RAL increased p62/SQSTM1 expression. Inhibition of TLR4, JNK, and PI3K/AKT blocked RAL-mediated increase of p62/SQSTM1. RAL activated JNK and PI3K/AKT signaling. RAL-mediated activations of JNK and PI3K/AKT signaling were reversed by TLR4 inhibition. Taken together, it is believed that RAL-mediated autophagy may be dependent on activating via TLR4-dependent activation of JNK and PI3K/AKT signaling in macrophages.

Temporal Transcriptome Analysis of SARS-CoV-2-Infected Lung and Spleen in Human ACE2-Transgenic Mice

  • Jung Ah, Kim;Sung-Hee, Kim;Jung Seon, Seo;Hyuna, Noh;Haengdueng, Jeong;Jiseon, Kim;Donghun, Jeon;Jeong Jin, Kim;Dain, On;Suhyeon, Yoon;Sang Gyu, Lee;Youn Woo, Lee;Hui Jeong, Jang;In Ho, Park;Jooyeon, Oh;Sang-Hyuk, Seok;Yu Jin, Lee;Seung-Min, Hong;Se-Hee, An;Joon-Yong, Bae;Jung-ah, Choi;Seo Yeon, Kim;Young Been, Kim;Ji-Yeon, Hwang;Hyo-Jung, Lee;Hong Bin, Kim;Dae Gwin, Jeong;Daesub, Song;Manki, Song;Man-Seong, Park;Kang-Seuk, Choi;Jun Won, Park;Jun-Won, Yun;Jeon-Soo, Shin;Ho-Young, Lee;Jun-Young, Seo;Ki Taek, Nam;Heon Yung, Gee;Je Kyung, Seong
    • Molecules and Cells
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    • v.45 no.12
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    • pp.896-910
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    • 2022
  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly transmissible and potentially fatal virus. So far, most comprehensive analyses encompassing clinical and transcriptional manifestation have concentrated on the lungs. Here, we confirmed evident signs of viral infection in the lungs and spleen of SARS-CoV-2-infected K18-hACE2 mice, which replicate the phenotype and infection symptoms in hospitalized humans. Seven days post viral detection in organs, infected mice showed decreased vital signs, leading to death. Bronchopneumonia due to infiltration of leukocytes in the lungs and reduction in the spleen lymphocyte region were observed. Transcriptome profiling implicated the meticulous regulation of distress and recovery from cytokine-mediated immunity by distinct immune cell types in a time-dependent manner. In lungs, the chemokine-driven response to viral invasion was highly elevated at 2 days post infection (dpi). In late infection, diseased lungs, post the innate immune process, showed recovery signs. The spleen established an even more immediate line of defense than the lungs, and the cytokine expression profile dropped at 7 dpi. At 5 dpi, spleen samples diverged into two distinct groups with different transcriptome profile and pathophysiology. Inhibition of consecutive host cell viral entry and massive immunoglobulin production and proteolysis inhibition seemed that one group endeavored to survive, while the other group struggled with developmental regeneration against consistent viral intrusion through the replication cycle. Our results may contribute to improved understanding of the longitudinal response to viral infection and development of potential therapeutics for hospitalized patients affected by SARS-CoV-2.