• Title/Summary/Keyword: Neutralizing antibodies

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Production of Characterization of Monoclonal Antibody to Glycoprotein D Antigen of Herpes simplex Virus Type 2

  • Choi, Young-Sook;Kim, Tae-Un;Lee, Inyung-Hoan;Cho, Myung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.173-178
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    • 2001
  • A monoclonal antibody (mAb) to the glucoprotein D (gD) of Herpes simplex virus type 2 (HSV-2) was successfully generated by hybridoma technology and characterized. The mAb, SKS2v, recognized a gD antigen with an apparent molecular mass of 60kDa in a Western blot analysis. The isotype was determined by a sandwich ELISA to be IgG2a. HSV-2 exhibited major antigens of 36, 43, 46, 47, 60, 69, 81, 96, 109, 112, 159, and 227 kDa among 25 protein profiles in SDS-PAGE, and among these antigens, those of 60, 112, 125, and 227 kDa were immunodominant in a Western blot analysis using antisera, thereby indicating that they play a role in inducing neutralizing antibodies in HSV-2 infection. When reacted with Vero cells infected with HSV-1 and HSV-2 SKSv2 showed a reactivity to the surface of the infected cells, and a gD antigen of 60 kDa appeared to be expressed in both types of HSV.

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AN INVESTIGATION OF IMMUNIZATION AGAINST SOMATOSTATIN BY MEASURING ANTIBODY TITRES, SOMATOSTATIN AND SOMATOTROPIN PROFILES IN GILTS

  • Du, Z.L.;Hacker, R.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.2
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    • pp.211-218
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    • 1993
  • The effect of active immunization against porcine somatostatin (SRIF-14) on somatostation and somatotropin secretion profile in 18 gilts was investigated. Gilts were assigned to the following treatments: control (sham injection, n = 6); bovine serum albumin (BSA) (injection of BSA with bacterial protein adjuvant, n = 6); SRIF (injection of BSA-SRIF-14 conjugate with bacterial protein adjuvant n = 6). Serum SRIF and pST were assayed from the blood samples taken on day 7 after the last immunization injection. Anti-SRIF antibody titres were assayed in weekly samples two weeks after the initial immunization to one week after the last immunization. Results revealed that the immunization protocol used in the present investigation failed to produce antibodies capable of neutralizing endogenous somatostatin. In addition, the porcine somatotropin assay revealed no significant differences in baseline pST concentration, mean peak amplitude and number of peaks during a 24 h secretory period among SRIF, BSA and control treatment. There were also no differences in SRIF baseline concentration, peak amplitude, and number of peaks during a 24 h secretory period among any of the three treatments. Circulating concentrations of pST and pSRIF were highly correlated (r = -0.09). Furthermore, anti-SRIF antibody titre was not detected in the serum of the gilts actively immunized against SRIF. These data, collectively, suggest that the protocol employed in the present investigation for active immunization against SRIF is not an effective method for changing SRIF and pST secretion profiles of the gilt and thus to enhance performance.

Prevalence on protective serum antibodies of canine distemper virus and canine parvovirus in Ulsan area (울산지역의 개 디스템퍼 및 파보 장염의 항체보유 실태 조사)

  • Sung, Ki-Chang;Lee, Eun-Woo;Park, Chang-Eun
    • Korean Journal of Veterinary Service
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    • v.33 no.3
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    • pp.213-221
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    • 2010
  • The results from a total of 412 blood samples consisted of 187 samples from regular visiting group (RV), 94 samples from first visiting group (FV), 52 samples from abandoned group (A), 54 samples from special breeder group (SB), and 25 samples from preliminary breeder group (PB) showed that RV(94.7%) and SB(88.9%) groups had the higher levels of protective antibody, but PB (36.0%) group revealed the lowest level. Among 96 blood samples with lower protective antibody levels, 14 samples (14.6%), 72 samples (75.0%) and 10 samples (10.4%) were below the protective antibody levels to distemper/parvo-virus, distemper only and parvovirus only, respectively. These results implied that antibody to parvovirus was well generated than that to distemper. Eighty six samples (20.9%) showed the protective antibody titer under 1:96 to distemper and 24 samples (5.8%), the protective antibody titer under 1:40 to parvovirus.

Discrimination of Hantaviruses from the Tissues of Infected Hamsters to 5 Different Serotype Hantaviruses by Nested RT-PCR using Hantavirus Serotype Specific Primers (한타바이러스 혈청형 특이 Primer를 이용한 Nested RT-PCR 방법으로 5가지 혈청형 한타바이러스에 감염된 햄스터 조직에서 바이러스 검출)

  • Chu, Yong-Kyu;Lee, Ho-Wang
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.49-57
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    • 1997
  • We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.

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Immunocytochemical identification of Aujeszky's disease viral antigens in nasal and circulating white blood cells from living pigs for early diagnosis (오제스키병의 생체 조기진단을 위한 돼지 비세포 및 순환백혈구내 면역세포화학적 바이러스 항원동정)

  • Moon, Oun-kyong;Cho, Hee-tack;Kim, Soon-bok;Sur, Jung-hyang
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.241-247
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    • 1993
  • Forty day old piglets were intranasally inoculated with 2ml of Aujeszky's disease virus (NYJ-1-87 strain, $10^{7.0}$ $TCID_{50/0.2}ml$), and the viral antigens were detected in nasal and circulating white blood cells for 20 days after inoculation by immunocytochemical method. Antibody titers in the blood were also detected by neutralizing test and Aujeszky's disease serodiagnostic kit(Choong Ang) in this periods. 1. Viral antigens were detected by the immunocytochemical technigue, and positive reactions were observated in nasal cells from the 2nd to the l0th days after inoculation and circulated white blood cells from the 4th to the 12th days after inoculation. 2. In neutralization test antibodies levels showed titers of 2 on the 8th day, 8 on the l5th day, 16 on the 18th day and 32 on the 20th day after inoculation. In serodiagnostic kit test positive reactions were observed after the 15th day after inoculation.

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Toll-Like Receptor-Mediated Free Radical Generation in Clonorchis sinensis Excretory-Secretory Product-Treated Cholangiocarcinoma Cells

  • Bahk, Young Yil;Pak, Jhang Ho
    • Parasites, Hosts and Diseases
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    • v.54 no.5
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    • pp.679-684
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    • 2016
  • Clonorchiasis, caused by direct contact with Clonorchis sinensis worms and their excretory-secretory products (ESPs), is associated with chronic inflammation, malignant changes in bile ducts, and even cholangiocarcinogenesis. Our previous report revealed that intracellular free radicals enzymatically generated by C. sinensis ESPs cause NF-${\kappa}B$-mediated inflammation in human cholangiocarcinoma cells (HuCCT1). Therefore, the present study was conducted to examine the role of upstream Toll-like receptors (TLRs) on the initial host innate immune responses to infection. We found that treatment of HuCCT1 cells with native ESPs induced changes in TLR mRNA levels in a time-dependent manner, concomitant with the generation of free radicals. ESP-mediated free radical generation was markedly attenuated by preincubation of the cells with TLR1-4-neutralizing antibodies, indicating that at least TLR1 through 4 participate in stimulation of the host innate immune responses. These findings indicate that free radicals triggered by ESPs are critically involved in TLR signal transduction. Continuous signaling by this pathway may function in initiating C. sinensis infection-associated inflammation cascades, a detrimental event leading to progression to more severe hepatobiliary diseases.

Immuno-Enhancing Effects through Macrophages of Polysaccharides Isolated from Citrus Peels (진피로부터 분리한 다당의 대식세포를 통한 면역증진 효과)

  • Lee, Kyung-Ae;Park, Hye-Ryung
    • The Korean Journal of Food And Nutrition
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    • v.34 no.5
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    • pp.441-448
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    • 2021
  • This study was designed to investigate the intracellular signaling pathways and immunoenhancing effect of macrophage activation by crude polysaccharides (CPP) extracted from citrus peels. CPP did not affect the cytotoxicity of RAW264.7 cells, but showed dose-dependent effects on cell viability. Also, CPP showed high production of chemokine (nitric oxide (NO)) and cytokines (interleukin (IL)-6 and tumor necrosis factor (TNF)-α). CPP increased IL-6, TNF-α, and inducible nitric oxide synthase (iNOS) mRNA expression dose-dependently. CPP also strongly induced the phosphorylation of the ERK, p38, and IκBα pathways in RAW 264.7 cells. In anti-pattern recognition receptors (PRRs) experiments, the effect of CPP on NO production was strongly suppressed by neutralizing toll-like receptor (TLR)2, TLR4, and Dectin1 antibodies, whereas IL-6 and TNF-α production by CPP was mainly suppressed by mannose receptor (MR). Therefore, these results suggest that CPP treatment-induced NO production was regulated by the ERK, p38, and NF-κB pathways through TLR2, TLR4, and Dectin1 receptors, whereas IL-6 and TNF-α production was primarily regulated by the ERK, p38, and NF-κB pathways through MR receptors.

Changes in age-specific seroprevalence of Japanese encephalitis virus and impact of Japanese encephalitis vaccine in Korea

  • Kwak, Byung Ok;Hong, Young Jin;Kim, Dong Hyun
    • Clinical and Experimental Pediatrics
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    • v.65 no.3
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    • pp.108-114
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    • 2022
  • The Japanese encephalitis (JE) virus is the leading cause of vaccine-preventable encephalitis in Asia. Since the introduction of a universal JE vaccination program and urbanization of Korea, the incidence of JE has dramatically decreased in Korea. However, recent JE cases have occurred, predominantly among unvaccinated adults and with a shift in age distribution. Here we aimed to review the changes in age-specific JE seroprevalence over time and discuss the implications of JE vaccination programs in Korea. Following the last epidemic in 1982-1983, mandatory vaccination for all children aged 3-15 years was conducted annually until 1994. However, JE has reemerged, predominantly affecting unvaccinated adults aged 40 years or older and demonstrating a shift in age distribution toward older populations. The age-specific seroprevalence of the JE virus in Korea has changed noticeably over time. Seropositivity in children and adolescents increased from 10%-59% in the 1970s to 90%-92% in the 1980s after the implementation of the JE vaccination program and increased further to 98% in 2012. No age-specific difference in the seroprevalence of JE was found, and appropriate levels of immunity to JE were maintained for all age groups. Continuous surveillance of the seroprevalence of JE is essential to establish a proper immunization policy in Korea.

Vaccine Strategy That Enhances the Protective Efficacy of Systemic Immunization by Establishing Lung-Resident Memory CD8 T Cells Against Influenza Infection

  • Hyun-Jung Kong;Youngwon Choi;Eun-Ah Kim;Jun Chang
    • IMMUNE NETWORK
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    • v.23 no.4
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    • pp.32.1-32.15
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    • 2023
  • Most influenza vaccines currently in use target the highly variable hemagglutinin protein to induce neutralizing antibodies and therefore require yearly reformulation. T cell-based universal influenza vaccines focus on eliciting broadly cross-reactive T-cell responses, especially the tissue-resident memory T cell (TRM) population in the respiratory tract, providing superior protection to circulating memory T cells. This study demonstrated that intramuscular (i.m.) administration of the adenovirus-based vaccine expressing influenza virus nucleoprotein (rAd/NP) elicited weak CD8 TRM responses in the lungs and airways, and yielded poor protection against lethal influenza virus challenge. However, a novel "prime-and-deploy" strategy that combines i.m. vaccination of rAd/NP with subsequent intranasal administration of an empty adenovector induced strong NP-specific CD8+ TRM cells and provided complete protection against influenza virus challenge. Overall, our results demonstrate that this "prime-and-deploy" vaccination strategy is potentially applicable to the development of universal influenza vaccines.

Multi-Immunogenic Outer Membrane Vesicles Derived from a MsbB-Deficient Salmonella enterica Serovar Typhimurium Mutant

  • Lee, Sang-Rae;Kim, Sang-Hyun;Jeong, Kang-Jin;Kim, Keun-Su;Kim, Young-Hyun;Kim, Sung-Jin;Kim, E-Kyune;Kim, Jung-Woo;Chang, Kyu-Tae
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1271-1279
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    • 2009
  • To develop low endotoxic and multi-immunogenic outer membrane vesicles (OMVs), a deletion mutant of the msbB gene in Salmonella enterica serovar Typhimurium (S. Typhimurium) was used as a source of low endotoxic OMV, and an expression vector of the canine parvovirus (CPV) VP2 epitope fused to the bacterial OmpA protein was constructed and transformed into the Salmonella ${\Delta}msbB$ mutant. In a lethality test, BALB/c mice injected intraperitoneally with the Salmonella ${\Delta}msbB$ mutant survived for 7 days, whereas mice injected intraperitoneally with the wild type survived for 3 days. Moreover, all mice inoculated orally with the ${\Delta}msbB$ mutant survived for 30 days, but 80% of mice inoculated orally with the wild type survived. The OmpA::CPV VP2 epitope fusion protein was expressed successfully and associated with the outer membrane and OMV fractions from the mutant S. Typhimurium transformed with the fusion protein-expressing vector. In immunogenicity tests, sera obtained from the mice immunized with either the Salmonella msbB mutant or its OMVs containing the OmpA::CPV VP2 epitope showed bactericidal activities against wild-type S. Typhimurium and contained specific antibodies to the CPV VP2 epitope. In the hemagglutination inhibition (HI) assay as a measurement of CPV-neutralizing activity in the immune sera, there was an 8-fold increase of HI titer in the OMV-immunized group compared with the control. These results suggested that the CPV-neutralizing antibody response was raised by immunization with OMV containing the OmpA::CPV VP2 epitope, as well as the protective immune response against S. Typhimurium in BALB/c mice.