• IMMUNE NETWORK
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• v.5 no.2
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• pp.68-77
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• 2005

Background: Costimulation is a critical process in Ag-specific immune responses. Both B7.1 and CD28 molecules have been reported to stimulate T cell responses during antigen presentation. Therefore, we tested whether Ag-specific immune responses as well as protective immunity are influenced by coinjecting with B7.1 and CD28 cDNAs in a mouse HSV-2 challenge model system. Methods: ELISA was used to detect levels of antibodies, cytokines and chemokines while thymidine incorporation assay was used to evaluate T cell proliferation levels. Results: Ag-specific antibody responses were enhanced by CD28 coinjection but not by B7.1 coinjection. Furthermore, CD28 coinjection increased IgG1 production to a significant level, as compared to pgD+pcDNA3, suggesting that CD28 drives Th2 type responses. In contrast, B7.1 coinjection showed the opposite, suggesting a Th1 bias. B7.1 coinjection also enhanced Ag-specific Th cell proliferative responses as well as production of Th1 type cytokines and chemokines significantly higher than pgD+pcDNA3. However, CD28 coinjection decreased Ag-specific Th cell proliferative responses as well as production of Th1 types of cytokines and chemokine significantly lower than pgD+pcDNA3. Only MCP-1 production was enhanced by CD28. B7.1 coimmunized animals exhibited an enhanced survival rate as well as decreased herpetic lesion formation, as compared to pgD+pcDNA3. In contrast, CD28 vaccinated animals exhibited decreased survival from lethal challenge. Conclusion: This study shows that B7.1 enhances protective Th1 type cellular immunity against HSV-2 challenge while CD28 drives a more detrimental Th2 type immunity against HSV-2 challenge, supporting an opposite role of B7.1 and CD28 in Ag-specific immune responses to a Th1 vs Th2 type.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.2
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• pp.244-249
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• 2004

The effects of supplemental betaine ($Betafin^{(R)}$) in the drinking water (50 g/kg) (WB) or feed (100 g/kg) (FB) were investigated on male broiler chickens ($Cobb{\times}Cobb$) exposed to 4 h episodes of heat stress at $34{\pm}1^{\circ}C$ on day (d) 35 and $36{\pm}1^{\circ}C$ from d 36 to 41. Prior to (d 1 to 34) and following heat exposure (d 35 to 41), betaine supplementation had no significant effect on body weight, total feed intake and cumulative feed conversion ratios of broilers. The total water intake of WB chicks was lower compared to controls. Prior to heat exposure, there was no difference in percentage of mortality among the three dietary groups. Following the heat challenge period, although higher percentage of control chicks succumbed to the heat challenge as compared to those of WB, it was not significantly different. The WB and FB chicks were less hyperthermic than controls in response to the heat challenge. Irrespective of treatment groups, the heat treatment resulted in a marked elevation in heterophil/lymphocyte ratios (HLR). The WB birds, however, had smaller increase in HLR than those of controls during heat exposure. Antibody production against Newcastle disease vaccine on day 35 was not affected by betaine supplementation. On d 42, WB birds had higher antibody production than those of FB. It is concluded that the WB treatment, as measured by HLR, antibody production and mortality rate, has advantages over the FB group under heat stress conditions.

• BMB Reports
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• v.44 no.11
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• pp.758-763
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• 2011

Immunostimulatory CpG-DNA targeting TLR9 is one of the most extensively evaluated vaccine adjuvants. Previously, we found that a particular form of natural phosphodiester bond CpG-DNA (PO-ODN) encapsulated in a phosphatidyl-${\beta}$-oleoyl-${\gamma}$-palmitoyl ethanolamine (DOPE) : cholesterol hemisuccinate (CHEMS) (1 : 1 ratio) complex (Lipoplex(O)) is a potent adjuvant. Complexes containing peptide and Lipoplex(O) are extremely useful for B cell epitope screening and antibody production without carriers. Here, we showed that IL-12 production was increased in bone marrow derived dendritic cells in a CpG sequence-dependent manner when PO-ODN was encapsulated in Lipoplex(O), DOTAP or lipofectamine. However, the effects of Lipoplex(O) surpassed those of PO-ODN encapsulated in DOTAP or lipofectamine and also other various forms of liposome-encapsulated CpG-DNA in terms of potency for protein antigen-specific IgG production and Th1- associated IgG2a production. Therefore, Lipoplex(O) may have a unique potent immunoadjuvant activity which can be useful for various applications involving protein antigens as well as peptides.

• Journal of Microbiology and Biotechnology
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• v.26 no.1
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• pp.190-196
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• 2016

In this study, the Brucella abortus ohr gene coding for an organic hydroperoxide resistance protein (Ohr) was cloned into a maltose fusion protein expression system (pMAL), inserted into Escherichia coli, and purified, and its immunogenicity was evaluated by western blot analysis using Brucella-positive mouse sera. The purified recombinant Ohr (rOhr) was treated with adjuvant and injected intraperitoneally into BALB/c mice. A protective immune response analysis revealed that rOhr induced a significant increase in both the IgG1 and IgG2a titers, and IgG2a reached a higher level than IgG1 after the second and third immunizations. Additionally, immunization with rOhr induced high production of IFN-γ as well as proinflammatory cytokines such as TNF, MCP-1, IL-12p70, and IL-6, but a lesser amount of IL-10, suggesting that rOhr predominantly elicited a cell-mediated immune response. In addition, immunization with rOhr caused a significantly higher degree of protection against a virulent B. abortus infection compared with a positive control group consisting of mice immunized with maltose-binding protein. These findings showed that B. abortus rOhr was able to induce both humoral and cell-mediated immunity in mice, which suggested that this recombinant protein could be a potential vaccine candidate for animal brucellosis.

• Korean Journal of Microbiology
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• v.43 no.1
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• pp.7-13
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• 2007

Pasteurella multocida is one of the important animal pathogen causing widespread infections in various domestic animals. In swine, it causes severe respiratory diseases such as atrophic rhinitis and pneumonic pasteurellosis. To develop the efficient subunit vaccine against swine atrophic rhinitis, we investigated protective antibodies and humoral immunity of outer membrane protein H (OmpH) which is one of the major outer membrane proteins in P. multocida. Outer membrane fraction of P. multocida was immunologically detectable using antisera from both mice groups vaccinated by formalin-killed whole cells and by commercial vaccine. The expression vector for production of recombinant OmpH was constructed and the recombinant OmpH was expressed and purified from E. coli. Recombinant OmpH showed high antigenic and immunogenic properties in mice vaccination and ELISA with antisera.

• Korean Journal of Poultry Science
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• v.35 no.3
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• pp.225-240
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• 2008

Marek's disease virus(MDV) is a highly cell-associated, lymphotropic $\alpha$-herpesvirus that causes paralysis and neoplastic disease in chickens. The disease has been controlled by vaccination which was provided the first evidence for a malignant cancer being controlled by an antiviral vaccine. Marek's disease pathogenesis is complex, involving cytolytic and latent infection of lymphoid cells and oncogenic transformation of $CD4^+$ T cells in susceptible chickens. MDV targets a number of different cell types during its life cycle. Lymphocytes play an essential role, although within them virus production is restricted and only virion are produced. Innate and adaptive immune responses develop in response to infection, but infection of lymphocytes results in immunosuppressive effects. Hence in MDV-infected birds, MDV makes its host more vulnerable to tumour development as well as to other pathogens. All chickens are susceptible to MDV infection, and vaccination is essential to protect the susceptible host from developing clinical disease. Nevertheless, MDV infects and replicates in vaccinated chickens, with the challenge virus being shed from the feather-follicle epithelium. The outcome of infection with MDV depends on a complex interplay of factors involving the MDV pathotype and the host genotype. Host factors that influence the course of MD are predominantly the responses of the innate and adaptive immune systems, and these are modulated by: age at infection and maturity of the immune system; vaccination status; the sex of the host; and various physiological factors.

• Journal of Microbiology and Biotechnology
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• v.22 no.11
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• pp.1580-1587
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• 2012

Japanese encephalitis virus (JEV) envelope (E) protein holds great promise for use in the development of a recombinant vaccine. Purified recombinant E (rE) protein may be useful for numerous clinical applications; however, there are limitations in using the Escherichia coli expression system for producing high-quality rE protein. Therefore, in this study, the yeast expression system was used to generate the rE protein. For protein production using the yeast system, the full-length JEV E gene was cloned into Pichia pastoris. SDS-PAGE and immunoblotting analysis demonstrated that the rE protein had a molecular mass of 58 kDa and was glycosylated. The predicted size of the mature unmodified E protein is 53 kDa, suggesting that post-translational modifications resulted in the higher molecular mass. The rE protein was purified to greater than 95% purity using combined ammonium sulfate precipitation and a SP-Sepharose Fast Flow column. This purified rE protein was evaluated for immunogenicity and protective efficacy in mice. The survival rates of mice immunized with the rE protein were significantly increased over that of Hyphantria cunea nuclear polyhedrosis virus E protein (HcE). Our results indicate that the rE protein expressed in the P. pastoris expression system holds great promise for use in the development of a subunit vaccine against JEV.

• Korean Journal of Veterinary Research
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• v.26 no.1
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• pp.97-102
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• 1986

The prevalence of diarrhea caused by enterotoxigenic Escherichia coli was surveyed on 445 calves in 6 farms which were located in the central part of Korea. The incidence of enterotoxigenic Escherichia coli in calves with diarrhea was investigated by detecting the K99 and F41 antigens from the isolated strains of Escherichia coli The incidence of colibacillosis in calves was 23.3%. Of 238 strains of Escherichia coli isolated from calves with diarrhea, 73 strains(30.6%) were proved possessing the K99 antigen by mannose-resistant hemagglutination(MRHA) using horse red blood cells and 79(33.1%) possessing F41 antigens by MRHA using guinea-pig red blood cells. The minca medium, nutrient broth, tryptic soy broth and brain heart infusion were tested for yield of K99 and F41 pili. The production of pili was greatest in minea medium. The best detachment method of the K99 and F41 pili from the cells was heat treatment for 20 minutes at $60^{\circ}C$ and concentration by precipitation with ammonium sulfate. The purified antigens of K99 and F41 were polypeptides with molecular weights of 18,500 and 29,500, respectively by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

• Korean Journal of Veterinary Service
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• v.35 no.3
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• pp.175-182
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• 2012

Rotaviruses have been known to be a major etiological agent of gastroenteritis in both infants and young animals. Subsequently new rotaviruses, which were morphologically indistinguishable but antigenically and electrophoretically distinct with each other, were reported from several animals throughout world including Korea. These new rotaviruses were named as non-group A or group B or group C rotaviruses and so on. It has been very difficult to isolate and grow the non-group A rotaviruses in vitro, and this has greatly limited the characterizations of non-group A rotaviruses and serological studies. In this study, monoclonal antibodies (MAbs) against porcine non-group A rotavirus were produced and characterized. The VP6 gene of porcine group C rotavirus Korean isolate(#06-52-1) was cloned and expressed. For expression of VP6 gene, baculovirus expression system was applied. The VP6 gene and expressed protein in the recombinant virus were confirmed by polymerase chain reaction (PCR), indirect fluorescence antibody (IFA) test and Western blot, respectively. The expressed VP6 was used for MAbs production. The MAbs produced in this study would be promising as diagnostic reagents for detection of group C rotavirus infection.

• Animal Bioscience
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• v.34 no.9
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• pp.1552-1558
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• 2021

Objective: This work was carried out to evaluate the effects of zilpaterol hydrochloride (ZH) and ractopamine hydrochloride (RH) combined with immunocastration on the welfare traits of feedlot Nellore cattle. Methods: Ninety-six Nellore males (average body weight [BW] = 409±50 kg; average 20 mo of age) were divided into two groups according to BW; half of the animals in each group received two doses of an immunocastration (ImC) vaccine in a 30 day interval, and the other half did not receive the vaccine (NoC). Afterward, the animals were housed and fed a common diet for 70 days. Then, they were split into three groups and fed one of the following diets for 30 additional days: control (CO) diet, with no β-AA; ZH diet, containing 80 mg/d ZH; and RH diet, containing 300 mg/d RH. Welfare traits were assessed by monitoring body surface temperature using infrared thermography (IRT) and plasma cortisol and temperament measurements. Results: There was no interaction between sexual condition and diet for any trait. The ImC and NoC groups did not differ in rectal and ocular temperatures. The ImC animals had higher flight speeds (p = 0.022) and tended to have higher cortisol levels (p = 0.059) than the NoC animals. Animals fed ZH and RH did not differ in cortisol levels, respiratory rate, rectal temperature, temperature measured by IRT, or temperament behaviour. Conclusion: The ImC animals showed a less stable temperament during handling practices than NoC, whereas ZH and RH supplementation had no adverse effects on animal welfare.