• Title/Summary/Keyword: two antigens

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Effect of Fc Fusion on Folding and Immunogenicity of Middle East Respiratory Syndrome Coronavirus Spike Protein

  • Chun, Jungmin;Cho, Yeondong;Park, Ki Hoon;Choi, Hanul;Cho, Hansam;Lee, Hee-Jung;Jang, Hyun;Kim, Kyung Hyun;Oh, Yu-Kyoung;Kim, Young Bong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.813-819
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    • 2019
  • Middle East respiratory syndrome coronavirus (MERS-CoV) induces severe respiratory impairment with a reported mortality rate of ~36% in humans. The absence of clinically available MERS-CoV vaccines and treatments to date has resulted in uncontrolled incidence and propagation of the virus. In vaccine design, fusion with the IgG Fc domain is reported to increase the immunogenicity of various vaccine antigens. However, limited reports have documented the potential negative effects of Fc fusion on vaccine antigens. To determine whether Fc fusion affects the immunogenicity of MERS-CoV antigen, we constructed a Fcassociated MERS-CoV spike protein (eS770-Fc, 110 kDa), whereby human IgG4 Fc domain was fused to MERS-CoV spike protein (eS770) via a Gly/Pro linker using baculovirus as the expression system. For comparative analyses, two eS770 proteins lacking the IgG4 Fc domain were generated using the IdeS protease ($eS770-{\Delta}Fc$) or His tag attachment (eS770-His) and the immunogenicity of the above constructs were examined following intramuscular immunization in mice. Contrary to expectations, non-Fc spike proteins ($eS770-{\Delta}Fc$, eS770-His; 90 kDa) showed higher immunogenicity than the Fc fusion protein (eS770-Fc). Moreover, unlike non-Fc spike proteins, eS770-Fc immunization did not elicit neutralizing antibodies against MERS-CoV. The lower immunogenicity of Fc-fused eS770 was related to alterations in the structural conformation of the spike protein. Taken together, our results indicate that IgG Fc fusion reduces the immunogenicity of eS770 by interfering with the proper folding structure.

Protective efficacy of a novel multivalent vaccine in the prevention of diarrhea induced by enterotoxigenic Escherichia coli in a murine model

  • Zhao, Hong;Xu, Yongping;Li, Gen;Liu, Xin;Li, Xiaoyu;Wang, Lili
    • Journal of Veterinary Science
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    • v.23 no.1
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    • pp.7.1-7.14
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    • 2022
  • Background: Enterotoxigenic Escherichia coli (ETEC) infection is a primary cause of livestock diarrhea. Therefore, effective vaccines are needed to reduce the incidence of ETEC infection. Objectives: Our study aimed to develop a multivalent ETEC vaccine targeting major virulence factors of ETEC, including enterotoxins and fimbriae. Methods: SLS (STa-LTB-STb) recombinant enterotoxin and fimbriae proteins (F4, F5, F6, F18, and F41) were prepared to develop a multivalent vaccine. A total of 65 mice were immunized subcutaneously by vaccines and phosphate-buffered saline (PBS). The levels of specific immunoglobulin G (IgG) and pro-inflammatory cytokines were determined at 0, 7, 14 and 21 days post-vaccination (dpv). A challenge test with a lethal dose of ETEC was performed, and the survival rate of the mice in each group was recorded. Feces and intestine washes were collected to measure the concentrations of secretory immunoglobulin A (sIgA). Results: Anti-SLS and anti-fimbriae-specific IgG in serums of antigen-vaccinated mice were significantly higher than those of the control group. Immunization with the SLS enterotoxin and multivalent vaccine increased interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations. Compared to diarrheal symptoms and 100% death of mice in the control group, mice inoculated with the multivalent vaccine showed an 80% survival rate without any symptom of diarrhea, while SLS and fimbriae vaccinated groups showed 60 and 70% survival rates, respectively. Conclusions: Both SLS and fimbriae proteins can serve as vaccine antigens, and the combination of these two antigens can elicit stronger immune responses. The results suggest that the multivalent vaccine can be successfully used for preventing ETEC in important livestock.

Individual expression and processing of hepatitis C virus E1/E2 epitopes-based DNA vaccine candidate in healthy humans' peripheral blood mononuclear cells

  • Rola Nadeem;Amany Sayed Maghraby;Dina Nadeem Abd-Elshafy;Ahmed Barakat Barakat;Mahmoud Mohamed Bahgat
    • Clinical and Experimental Vaccine Research
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    • v.12 no.1
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    • pp.47-59
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    • 2023
  • Purpose: The development and study of hepatitis C virus (HCV) vaccine candidates' individualized responses are of great importance. Here we report on an HCV DNA vaccine candidate based on selected envelope (E1/E2) epitopes. Besides, we assessed its expression and processing in human peripheral blood mononuclear cells (PBMCs) and in vivo cellular response in mice. Materials and Methods: HCV E1/E2 DNA construct (EC) was designed. The antigen expression of EC was assayed in PBMCs of five HCV-uninfected donors via a real-time quantitative polymerase chain reaction. Serum samples from 20 HCV antibody-positive patients were used to detect each individual PBMCs expressed antigens via enzyme-linked immunosorbent assay. Two groups, five Swiss albino mice each, were immunized with the EC or a control construct. The absolute count of lymph nodes' CD4+ and CD8+ T-lymphocytes was assessed. Results: Donors' PBMCs showed different levels of EC expression, ranging between 0.83-2.61-fold in four donors, while donor-3 showed 34.53-fold expression. The antigens expressed in PBMCs were significantly reactive to the 20 HCV antibody repertoire (all p=0.0001). All showed comparable reactivity except for donor-3 showing the lowest reactivity level. The absolute count % of the CD4+ T-cell significantly increased in four of the five EC-immunized mice compared to the control group (p=0.03). No significant difference in CD8+ T-cells % was observed (p=0.89). Conclusion: The inter-individual variation in antigen expression and processing dominance was evident, showing independence in individuals' antigen expression and reactivity levels to antibodies. The described vaccine candidate might result in a promising natural immune response with a possibility of CD4+ T-cell early priming.

Diagnostic Value of p53 Expression in the Evaluation of Effusions (체강삼출액의 진단에 있어서 p53 단백의 유용성)

  • Lee, Ji-Shin;Park, Chang-Soo
    • The Korean Journal of Cytopathology
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    • v.7 no.2
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    • pp.138-143
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    • 1996
  • The diagnostic accuracy of routine cytological preparations from effusions ranges from 60% to 70%. Immunohistochemical markers, especially tumor-associated antigens, have been successfully employed to increase diagnostic sensitivity in effusion cytology. However, more than two different antibodies in diagnosis of effusions are needed. In the view of prevalence of abnormalities of p53 gene in human malignancies we investigated the diagnostic usefulness of demonstration of p53 protein immunoreactivity in distinguishing benign changes versus malignant processes in effusions. p53 protein expression was studied immunohistochemically in 76 effusions(28 malignant and 48 benign) using anti-human p53 antibody p53 immunoreactivity was identified in 19 of 28(67.9%) malignant effusions. In contrast, no p53 immunoreactivity was observed in all benign effusions. A specificity of 100% and a sensitivity of 67.9% were observed. These results suggest that immunohistochemical detection of p53 protein seems to be helpful in distinguishing benign changes versus malignant processes in effusions, although its principal limitation is its relatively low sensitivity.

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Recent Advances in the Innate Immunity of Invertebrate Animals

  • Iwanaga, Sadaaki;Lee, Bok-Luel
    • BMB Reports
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    • v.38 no.2
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    • pp.128-150
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    • 2005
  • Invertebrate animals, which lack adaptive immune systems, have developed other systems of biological host defense, so called innate immunity, that respond to common antigens on the cell surfaces of potential pathogens. During the past two decades, the molecular structures and functions of various defense components that participated in innate immune systems have been established in Arthropoda, such as, insects, the horseshoe crab, freshwater crayfish, and the protochordata ascidian. These defense molecules include phenoloxidases, clotting factors, complement factors, lectins, protease inhibitors, antimicrobial peptides, Toll receptors, and other humoral factors found mainly in hemolymph plasma and hemocytes. These components, which together compose the innate immune system, defend invertebrate from invading bacterial, fungal, and viral pathogens. This review describes the present status of our knowledge concerning such defensive molecules in invertebrates.

An Optimal Parameter Selection of Power System Stabilizer using Immune Algorithm (면역 알고리즘을 이용한 전력 계통 안정화 장치의 최적 파라미터 선정)

  • Jeong, Hyeong-Hwan;Lee, Jeong-Pil;Jeong, Mun-Gyu;Lee, Gwang-U
    • The Transactions of the Korean Institute of Electrical Engineers A
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    • v.49 no.9
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    • pp.433-445
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    • 2000
  • In this paper, optimal tuning problem of power system stabilizer(PSS) using Immune Algorithm(IA) is investigated to improve power system dynamic stability. In proposed method, objective function is represented as antigens. An affinity calculation is embedded within the algorithm for determining the promotion or suppression of antibody. An antibody that most fits the antigen is considered as the solution to PSS tuning problem. The computaton performance by the proposed method is compared with Genetic Algorithm(GA). The porposed PSS using IA has been applied for two sample system, single-machine infinite bus system and multi-machine power system. The performance of the proposed PSS is compared with that of conventional PSS. It is shown that the proposed PSS tuned using immune algorithm is more robust than conventional PSS.

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Rheological Property and Adjuvant Effect of l-Carrageenan (l-카라기난의 유동학적 성질과 어쥬번트 효과)

  • 김하형;김범수;이광재;이종혁;조두현;김윤중;정병욱
    • YAKHAK HOEJI
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    • v.46 no.1
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    • pp.6-10
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    • 2002
  • The present study investigated the rheological properties and adjuvant activity of IgG production of ι-Carrageenan prepared using two different methods . incubation at 37.4$^{\circ}C$ after either being heated or not being heated int 6$0^{\circ}C$ for 10 minutes. The carrageenan exhibited plastic flow with thixotropy in both cases, but the viscosity of the sample that had been heated was significantly higher. C57BL/6 mice were then immunized intraperitoneally with either of the carrageenans containing protein antigens. The maximum adjuvant effect was found when using the nonheated carrageenan, especially in term of IgG2a, IgG2b, and IgG3 production. In contrast, IgG1 antibody production was the same far both types of carrageenan. furthermore, when administered oral1y, neither type of carrageenan resulted in the production of any antibodies.

Activation-induced Cytidine Deaminase in B Cell Immunity and Cancers

  • Park, Seok-Rae
    • IMMUNE NETWORK
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    • v.12 no.6
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    • pp.230-239
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    • 2012
  • Activation-induced cytidine deaminase (AID) is an enzyme that is predominantly expressed in germinal center B cells and plays a pivotal role in immunoglobulin class switch recombination and somatic hypermutation for antibody (Ab) maturation. These two genetic processes endow Abs with protective functions against a multitude of antigens (pathogens) during humoral immune responses. In B cells, AID expression is regulated at the level of either transcriptional activation on AID gene loci or post-transcriptional suppression of AID mRNA. Furthermore, AID stabilization and targeting are determined by post-translational modifications and interactions with other cellular/nuclear factors. On the other hand, aberrant expression of AID causes B cell leukemias and lymphomas, including Burkitt's lymphoma caused by c-myc/IgH translocation. AID is also ectopically expressed in T cells and non-immune cells, and triggers point mutations in relevant DNA loci, resulting in tumorigenesis. Here, I review the recent literatures on the function of AID, regulation of AID expression, stability and targeting in B cells, and AID-related tumor formation.

Rotavirus Vaccines (로타바이러스 백신)

  • Koh, Hong
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.12 no.sup1
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    • pp.72-76
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    • 2009
  • Rotavirus infection is the leading cause of severe diarrhea disease in infants and young children worldwide. Rotavirus infects every child at least once by her/his $5^{th}$ birthday. It has been known that single episode of rotavirus infection can protect or alleviate subsequent illness caused by both homotypic and heterotypic rotaviruses. There are two currently licensed rotavirus vaccines. One is human-bovine rotavirus reassortant pentavalent vaccine ($RotaTeq^{TM}$), which contains five reassortant rotavirus (expressing protein G1, G2, G3, G4 and P[8]) and was licensed in Korea for use among infants in 2007. Another is live-attenuated human rotavirus vaccine ($Rotarix^{TM}$) derived from 89-12 strain which represents the most common of the human rotavirus VP7(G1) and VP4(P[8]) antigens. $Rotarix^{TM}$ was licensed in Korea in 2008. Both live oral rotavirus vaccines are efficacious in preventing severe rotavirus gastroenteritis.

Convenient Preparation of Tumor-specific Immunoliposomes Containing Doxorubicin

  • Nam, Sang-Min;Cho, Jang-Eun;Son, Byoung-Soo;Park, Yong-Serk
    • BMB Reports
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    • v.31 no.1
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    • pp.95-100
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    • 1998
  • Two innovative methods to prepare target-sensitive immunoliposomes containing doxorubicin by coupling monoclonal antibodies (mAb DH2, SH1) specific to cancer cell surface antigens ($G_{M3}$, $Le^X$) have been developed and are described here. Firstly, liposomes containing N-glutaryl phosphatidylethanolamine (NGPE) were prepared, followed by the encapsulation of doxorubicin, DH2 or SH1 antibodies were conjugated to NGPE in the liposomes (direct coupling). Secondly, liposomes were prepared with NGPE/mAb conjugates by the detergent dialysis method (conjugate insertion), and then doxorubicin was encapsulated by proton gradient. The immunoliposomes prepared by both methods were able to specifically bind to the surface of the tumor cells - B16BL6 mouse melanoma cells. The efficiencies of doxorubicin-entrapping into liposomes prepared by direct coupling and conjugate insertion was about 98% and 25%, respectively. These types of liposomal formulation are sensitive to target cells, which can be useful for various clinical applications.

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