• Journal of Veterinary Science
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• 제21권5호
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• pp.74.1-74.13
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• 2020

Background: The quality of a vaccine depends strongly on the effects of the adjuvants applied simultaneously with the antigen in the vaccine. The adjuvants enhance the protective effect of the vaccine against a viral challenge. Conversely, oil-type adjuvants leave oil residue inside the bodies of the injected animals that can produce a local reaction in the muscle. The long-term immunogenicity of mice after vaccination was examined. ISA206 or ISA15 oil adjuvants maintained the best immunity, protective capability, and safety among the oil adjuvants in the experimental group. Objectives: This study screened the adjuvant composites aimed at enhancing foot-and-mouth disease (FMD) immunity. The C-type lectin or toll-like receptor (TLR) agonist showed the most improved protection rate. Methods: Experimental vaccines were fabricated by mixing various known oil adjuvants and composites that can act as immunogenic adjuvants (gel, saponin, and other components) and examined the enhancement effect on the vaccine. Results: The water in oil (W/O) and water in oil in water (W/O/W) adjuvants showed better immune effects than the oil in water (O/W) adjuvants, which have a small volume of oil component. The W/O type left the largest amount of oil residue, followed by W/O/W and O/W types. In the mouse model, intramuscular inoculation showed a better protection rate than subcutaneous inoculation. Moreover, the protective effect was particularly weak in the case of inoculation in fatty tissue. The initial immune reaction and persistence of long-term immunity were also confirmed in an immune reaction on pigs. Conclusions: The new experimental vaccine with immunostimulants produces improved immune responses and safety in pigs than general oil-adjuvanted vaccines.

• Journal of Microbiology and Biotechnology
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• 제27권4호
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• pp.718-724
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• 2017

The combination of rabies immunoglobulin (RIG) with a vaccine is currently effective against rabies infections, but improvements are needed. Genetic engineering antibody technology is an attractive approach for developing novel antibodies to replace RIG. In our previous study, a single-chain variable fragment, scFv57R, against rabies virus glycoprotein was constructed. However, its inherent weak stability and short half-life compared with the parent RIG may limit its diagnostic and therapeutic application. Therefore, an acidic tail of synuclein (ATS) derived from the C-terminal acidic tail of human alpha-synuclein protein was fused to the C-terminus of scFv57R in order to help it resist adverse stress and improve the stability and half-life. The tail showed no apparent effect on the preparation procedure and affinity of the protein, nor did it change the neutralizing potency in vitro. In the ELISA test of molecular stability, the ATS fusion form of the protein, scFv57R-ATS, showed an increase in thermal stability and longer half-life in serum than scFv57R. The protection against fatal rabies virus challenge improved after fusing the tail to the scFv, which may be attributed to the improved stability. Thus, the ATS fusion approach presented here is easily implemented and can be used as a new strategy to improve the stability and half-life of engineered antibody proteins for practical applications.

• KSBB Journal
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• 제19권1호
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• pp.88-92
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• 2004

장티푸스 협막 다당체와 일본 뇌염 바이러스로 구성되어있는 혼합백신을 제조하여 마우스에서 면역성을 측정하였다. 혼합 항원 백신은 단일 항원을 투여한 경우보다 장티푸스와 일본 뇌염에 대해서 더 높은 IgG 항체 형성을 나타내었다. Aluminium hydroxide를 adjuvant로 첨가한 경우에도 첨가하지 않은 대조군에 비하며 모두 IgG 항체 형성이 증가하는 것으로 나타났다. 일본뇌염의 경우 중화항체 값 측정 시험에서도 혼합 항원백신은 일본뇌염 단독 항원 시에 비교해서 더 많은 중화항체 값을 형성하는 결과를 나타내었다. 본 실험에서 장티푸스 협막 다당체와 일본뇌염 바이러스의 혼합백신은 서로 다른 성분의 항원간에 나타날 수 있는 항체 형성에 대한 masking effect가 발생하지 않으며 오히려 synergic effect를 나타낸다는 사실을 보여주었다. 본 연구에서는 접종시기가 유사한 두 항원의 혼합 백신으로의 사용가능성을 높여주고 최근에 백신 산업에서 종류가 다른 여러 백신들을 혼합하여 다가백신을 제조 생산하는 추세의 산업화에 큰 기대효과를 제공할 수 있으리라 사려된다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제4권1호
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• pp.66-71
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• 1999

One of the practical limitations with the use of liposomes for delivery of the pharmaceutical substances such antigens is that liposomes are relatively unstable in storage. In order to extend the stability of liposome in storage without affecting their functional activity, solution-type liposomes were dehydrated to form a structurally intact dry liposomes. Comparative immunological evaluation was carried out for both dry and solution-type liposomes containing gag-V3 chimera, consequently it was found that dry liposomes elicited both humoral and cellular response as efficiently as solution-type liposemes did against the same gag-V3 antigen. Especially, long-term stability of the liposomes was remarkably enhanced by the dehydration made to loposomes without a significant change in its ability to elicit immune response in vivo. These results indicate that dry pH-sensitive liposome may become an effective delivery and adjuvant system for general vaccine development.

• Biomolecules & Therapeutics
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• 제7권1호
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• pp.89-96
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• 1999

CJ-50002 is an oral vaccine against V.vulnificus infection composed of whole cell lysate of V. vulnificus. The general pharmacological properties of CJ-50002 were evaluated in various animals and in vitro system. CJ-50002 at oral doses of 0.2, 2 and 20 mg/kg had no effect on general behavior in mice, chromo- and electro-convulsions in mice, writhing syndrome induced by acetic acid in mice, body temperature in rats, charcoal meal propulsion in mice and urine and electrolytes excretion in rats. However, oral administration of CJ-50002 at dose of 20 mg/kg prolonged the hexobarbital-inuced sleeping inducing time in mice. In anesthetized dogs, CJ-50002 showed no effect on blood pressure, heart rate and ECG but decreased the respiratory rate and femoral blood flow at dose of 20 mg/kg. p.o. CJ-50002 had no effect on the contractile response of the isolated guinea pig ileum to various spasmogen at concentrations of 0.2, 2 and 20 $\mu\textrm{g}$/ml, respectively. Since these pharmacological effects of CJ-500o2 were observed at dose much greater than those in clinical use (approximately 0.16 mg/kg, p.o.), it is likely that this vaccine may be relatively free of undesirable effects in clinical practice.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2006년도 제23차 정기총회 및 학술발표회
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• pp.90-91
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• 2006

To reduce the economic impact and control Low pathogenic avian influenza (LPAI), vaccination with inactivated vaccine has been considered in this country. We tried to develop inactivated vaccine with reassorted H9N3 AI virus which has different type of neuraminidase compare to those of field AI virus. Before reassorted vaccine was produced, we confirm the virus as master seed by limiting dilution, RT-PCR and sequencing method. Also, we evaluate the biological characteristics of the virus to find out the possibility of prevention against field infection of AI virus. Finally, we evaluate the safety and efficacy of the vaccine made of reassorted AI virus in the specific pathogen free (SPF) chickens. After limiting dilution, we choose RV7CE4 as a vaccine candidate and compare the gene sequence of this vaccine strain to those of AI05GA which is parents strain. Compared to amino acid sequences of specific gene of AI05GA and RV7CE4, exhibited a high degree of amino acid sequence homology. In the safety and efficacy test, there were no specific clinical signs or mortality. Reassorted H9N3 viruses were reisolated in cloaca swab on 5 days post inoculation. In the vaccine study, once or twice vaccination was performed and challenged with H9N2 field virus (01310). Vaccine has no adverse effect on birds and formed good immune capability which reduce viral shedding in the birds infected with 01310. Based on the above result, we developed reassorted H9N3 vaccine which will efficiently prevent the low pathogenic AIV (H9N2) infection in the poultry farms.

• IMMUNE NETWORK
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• 제3권2호
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• pp.110-117
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• 2003

Background: The usefulness of DNA vaccine at priming step of heterologous prime-boost vaccination led to DNA vaccine closer to practical reality. DNA vaccine priming followed by recombinant viral vector boosting via systemic route induces optimal systemic immunity but no mucosal immunity. Mucosal vaccination of the reversed protocol (recombinant viral vector priming-DNA vaccine boosting), however, can induce both maximal mucosal and systemic immunity. Here, we tried to address the reason why the mucosal protocol of prime-boost vaccination differs from that of systemic vaccination. Methods: To address the importance of primary immunity induced at priming step, mice were primed with different doses of DNA vaccine or coadministration of DNA vaccine plus mucosal adjuvant, and immunity including serum IgG and mucosal IgA was then determined following boosting with recombinant viral vector. Next, to assess influence of humoral pre-existing immunity on boosting $CD8^+$ T cell-mediated immunity, $CD8^+$ T cell-mediated immunity in B cell-deficient (${\mu}K/O$) mice immunized with prime-boost regimens was evaluated by CTL assay and $IFN-{\gamma}$-producing cells. Results: Immunity primed with recombinant viral vector was effectively boosted with DNA vaccine even 60 days later. In particular, animals primed by increasing doses of DNA vaccine or incorporating an adjuvant at priming step and boosted by recombinant viral vector elicited comparable responses to recombinant viral vector primed-DNA vaccine boosted group. Humoral pre-existing immunity was also unlikely to interfere the boosting effect of $CD8^+$ T cell-mediated immunity by recombinant viral vector. Conclusion: This report provides the important point that optimally primed responses should be considered in mucosal immunization of heterologous prime-boost regimens for inducing the effective boosting at both mucosal and systemic sites.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.1022-1029
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• 2018

Tuberculosis (TB) remains a serious health issue around the word. Adenovirus (Ad)-based vaccine and modified vaccinia virus Ankara (MVA)-based vaccine have emerged as two of the most promising immunization candidates over the past few years. However, the performance of the homologous and heterologous prime-boost immunization regimens of these two viral vector-based vaccines remains unclear. In the present study, we constructed recombinant Ad and MVA expressing an Ag85B-TB10.4 fusion protein (AdH4 and MVAH4) and evaluated the impact of their different immunization regimens on the humoral and cellular immune responses. We found that the viral vector-based vaccines could generate significantly higher levels of antigen-specific antibodies, $IFN-{\gamma}$-producing splenocytes, $CD69^+CD8^+$ T cells, and $IFN-{\gamma}$ secretion when compared with bacillus Calmette-$Gu{\acute{e}}rin$ (BCG) in a mouse model. AdH4-containing immunization regimens (AdH4-AdH4, AdH4-MVAH4, and MVAH4-AdH4) induced significantly stronger antibody responses, much more $IFN-{\gamma}$-producing splenocytes and $CD69^+CD8^+$ T cells, and higher levels of $IFN-{\gamma}$ secretion when compared with the MVAH4-MVAH4 immunization regimen. The number of $IFN-{\gamma}$-producing splenocytes sensitive to $CD8^+$ T-cell restricted peptides of Ag85B (9-1p and 9-2p) and Th1-related cytokines ($IFN-{\gamma}$ and $TNF-{\alpha}$) in the AdH4-MVAH4 heterologous prime-boost regimen immunization group was significantly higher than that in the other viral vector-based vaccine- and BCG-immunized groups, respectively. These results indicate that an immunization regimen involving AdH4 may have a higher capacity to induce humoral and cellular immune responses against TB in mice than that by regimens containing BCG or MVAH4 alone, and the AdH4-MVAH4 prime-boost regimen may generate an ideal protective effect.

• 약학회지
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• 제59권1호
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• pp.1-5
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• 2015

Cordyceps militaris has shown to have various pharmacological activities including an immune-modulatory effect. Previously, we reported that anti-influenza effect of C. militaris in DBA/2 mice was mediated by increased IL-12 and the activation of NK cells. In this study, we investigated the effect of C. militaris on adaptive immune responses using DBA2 mice immunized with influenza vaccine. To determine the effect of C. militaris on antigen presentation capability, we treated RAW 264.7 cells with various concentrations of ethanol extract of C. militaris, which showed a significant upregulation of CD86 (B7.2), CD284 (TLR4), CD40, H-2k (MHC I) and I-Ad (MHC II). To examine the direct effect of C. militaris on adaptive immune responses, we immunized DBA2 mice with influenza vaccine in presence or absence of C. militaris. After 2 or 4 weeks, influenza-specific T cell proliferation, HAI titers and IFN-${\gamma}$ production were measured in plasma or PBMCs isolated from animals. Influenza-specific T cell proliferation and HAI titers were not considerably increased in immunized mice in presence of C. militaris. However, the production of IFN-${\gamma}$ was much greater in immunized mice with C. militaris as adjuvant than only immunized mice.

• 한국동물위생학회지
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• 제35권3호
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• pp.169-174
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• 2012

Japanese encephalitis virus (JEV) is one of the main causes of viral encephalitis in human and animals. For over 30 years, a live attenuated JEV vaccine strain has been used in the veterinary field, and it is required to conduct quality evaluation studies on the commercial vaccines. For the quality control of live attenuated JEV vaccine, we investigated the nucleotide sequence similarity of prME gene derived from five JEV vaccines commercially available in pigs in Korea. The Vero cells infected with JEV vaccines showed specific cytopathic effect, which was characterized by rounding and detached cells. In the phylogenetic analysis, all of the vaccine strains showed a close relationship with the original vaccine seed strain (Anyang 300) and clustered into the genotype 3. In comparison of the nucleotide and deduced amino acid sequences of prME genes with the original strain, all JEV vaccine strains showed high amino acid similarity ranging from 98.9% to 99.5%, but had several point mutations, probably due to high mutation rates of viral RNA polymerase by several virus passages. Even though the current JEV vaccine strains have been maintained and produced for a long period of time, the genetic characterization of them have been rarely changed. However, since the mid 1990's, molecular epidemiology of JEV has been changed sharply from genotype 3 to genotype 1 in Korea, further studies on new vaccine strains to genotype 1 is required for more effective prevention in the field.