• Journal of fish pathology
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• v.27 no.1
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• pp.47-56
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• 2014

Propolis is a beehive product with a very complex chemical composition, widely used in folk medicine because of its several therapeutic activities. This study was conducted to measure the efficacy of propolis on non-specific defense reactions, specific immune response, and protection levels against pathogen challenge with Streptococcus iniae. in vitro and in vivo. In vitro, the phagocytic activity and NBT assay of peripheral blood leukocytes (PBL) were evaluated in a various propolis extractsconcentrations (0, 10, 50, 100, 150, 250 and $500{\mu}g/ml$). The optimal concentration showing activation of propolis extracts was determined to $100{\mu}g/ml$. In vivo, they were divided into four groups (PBS, propoli extractss, vaccine, propolis extracts + vaccine) in vivo. Fish were received i.p. injection of either PBS or propolis extracts, and in the presence or absence of formalin inactivated S. iniae ($1{\times}10^8$ CFU/fish), respectively. The level of haematocrit is not affected among experimental groups. The phagocytic activity and the NBT reduction activities of head kidney phagocyte were markedly (p<0.05)augmented in the propolis extracts groups than in the PBS-control group, respectively. The level of serum lysozyme activity was significantly (p<0.05) increased in the propolis extracts treated groups than in the PBS-control group. The agglutinin titer was significantly (p<0.05) enhanced in the vaccine+propolis extracts group than in the vaccine group, but there was no difference between PBS-control and propolis treated group. The results of the present study suggest that propolis extracts seems to be a promising compounds of non-specific immune stimulator, also being able to use a good adjuvant.

• Korean Journal of Veterinary Research
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• v.52 no.2
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• pp.89-97
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• 2012

Mycobacterial cell-wall skeleton (CWS) is an immunoactive and biodegradable particulate adjuvant and has been tried to use for immunotherapy. The CWS of Mycobacterium bovis bacillus Calmette-Guerin (BCG-CWS) was studied as an universal vaccine vehicle for antigen conjugation, to develop potentially effective and safe vaccine. Although a variety of biological activities of BCG-CWS have been studied, the effects of BCG-CWS on spleen cells are not fully elucidated. Using MTT assay and trypan blue exclusion test, we found that BCG-CWS significantly enhanced the viability and proliferation of cells. Multiple clusters, indicating proliferation, were observed in BCG-CWS-treated spleen cells and surface marker staining assay revealed that BCG-CWS promoted the proliferation of $CD19^+$ B lymphocyte rather than $CD4^+$ or $CD8^+$ T lymphocyte. In addition, BCG-CWS up-regulated the expression of anti-apoptotic molecules such as bcl-2, bcl-xL. BCG-CWS increased the surface expression of CD25 and CD69 as well as IL-2 production of spleen cells, suggesting increased activation. Furthermore, BCG-CWS enhanced the antigen-specific cell proliferation and interferon-gamma production of spleen cells. Taken together, these results demonstrate the immunostimulatory effects of BCG-CWS on spleen cells via multiple mechanisms, providing valuable information to broaden the use of BCG-CWS in clinical and research settings.

• Journal of Microbiology and Biotechnology
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• v.31 no.2
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• pp.304-316
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• 2021

Vaccination is the most effective way to prevent influenza virus infections. However, conventional vaccines based on hemagglutinin (HA) have to be annually updated because the HA of influenza viruses constantly mutates. In this study, we produced a 3M2e-3HA2-NP chimeric protein as a vaccine antigen candidate using an Escherichia coli expression system. The vaccination of chimeric protein (15 ㎍) conferred complete protection against A/Puerto Rico/8/1934 (H1N1; PR8) in mice. It strongly induced influenza virus-specific antibody responses, cytotoxic T lymphocyte activity, and antibody-dependent cellular cytotoxicity. To spare the dose and enhance the cross-reactivity of the chimeric, we used a complex of poly-γ-glutamic acid and alum (PGA/alum) as an adjuvant. PGA/alum-adjuvanted, low-dose chimeric protein (1 or 5 ㎍) exhibited higher cross-protective effects against influenza A viruses (PR8, CA04, and H3N2) compared with those of chimeric alone or alum-adjuvanted proteins in vaccinated mice. Moreover, the depletion of CD4+ T, CD8+ T, and NK cells reduced the survival rate and efficacy of the PGA/alum-adjuvanted chimeric protein. Collectively, the vaccination of PGA/alum-adjuvanted chimeric protein induced strong protection efficacy against homologous and heterologous influenza viruses in mice, which suggests that it may be a promising universal influenza vaccine candidate.

• Journal of Veterinary Science
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• v.24 no.1
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• pp.15.1-15.13
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• 2023

Background: Inactivated vaccines are limited in preventing foot-and-mouth disease (FMD) due to safety problems. Recombinant virus-like particles (VLPs) are an excellent candidate for a novel vaccine for preventing FMD, given that VLPs have similar immunogenicity as natural viruses and are replication- and infection-incompetent. Objectives: The 3C protease and P1 polyprotein of type O FMD virus (FDMV) was expressed in yeast Hansenula polymorpha to generate self-resembling VLPs, and the potential of recombinant VLPs as an FMD vaccine was evaluated. Methods: BALB/c mice were immunized with recombinant purified VLPs using CpG oligodeoxynucleotide and aluminum hydroxide gel as an adjuvant. Cytokines and lymphocytes from serum and spleen were analyzed by enzyme-linked immunosorbent assay, enzyme-linked immunospot assay, and flow cytometry. Results: The VLPs of FMD were purified successfully from yeast protein with a diameter of approximately 25 nm. The immunization of mice showed that animals produced high levels of FMDV antibodies and a higher level of antibodies for a longer time. In addition, higher levels of interferon-γ and CD4⁺ T cells were observed in mice immunized with VLPs. Conclusions: The expression of VLPs of FMD in H. polymorpha provides a novel strategy for the generation of the FMDV vaccine.

• Journal of Pharmaceutical Investigation
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• v.34 no.1
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• pp.1-14
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• 2004

A promising approach to the development of a new single-step vaccine, which would eliminate the requirement for multiple injections, involves the encapsulation of antigens into microspheres. Biodegradable poly(lactide-co-glycolide) (PLGA) microspheres gave us a bright insight for controling antigen release in a pulsatile fashion, thereby mimicking two or tree boosting injections. However, in spite of the above merits, the level of immunization induced by a single-shot vaccination is often lower tan two doses of alum-adsorbed antigen. Therefore, optima modification of the microsphere is essential for the development of single-step vaccines. In the review, we discuss the stability of antigen in microsphere, safety and non-toxic in human and encapsulation technology. Also, we attempted to outline relevant physicochemical properties on the immunogenicity of microsphere vaccine and attainment of pulsatile release pater by combination of different microsphere, as well as to analyze immunological data associated with antigen delivery by microsphere. Although a lot of variables are related to the optimized microsphere formulation, we could conclude that judicious choice of proper polymer type, adjustment of particles size, and appropriate immunization protocol along with a suitable adjuvant might be a crucial factor for the generation of long-lasting immune response from a single-step vaccine formulation employing PLGA microsphere.

• Korean Journal of Veterinary Service
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• v.36 no.1
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• pp.1-6
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• 2013

Since the 1980's, several kinds of inactivated bovine viral diarrhea virus (BVDV) vaccines have been used to immunize domestic animals such as cattle and goat in Korea. Immunogenicity of the BVDV vaccines has been checked by the Korean Veterinary Authority using laboratory animals. In this study, we applied a molecular method to investigate the genetic characterization of the BVDV genes in six commercial inactivated BVDV vaccines, and determined the efficiency of two extraction reagents (i.e., sodium citrate or isopropyl myristate) to separate the vaccine antigens from the antigen/adjuvant complexes. Six partial non-coding regions (288 bp) were successfully amplified with specific primer sets, which demonstrated that sodium citrate is more efficient in extracting viral RNA from inactivated gel vaccines than isopropyl myristae. In addition, we identified the virus strains from the vaccines by analyzing the nucleotide sequences of the 5' non-coding region (NCR) of BVDV. The nucleotide similarity of the partial 5' NCR ranged from 95.1 to 100% among BVDV vaccine strains, respectively, indicating that a few manufacturers used different BVDV strains to produce their vaccines.

• Biomolecules & Therapeutics
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• v.2 no.4
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• pp.331-335
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• 1994

As a part of the safety evaluation of Pseudomonas vaccine(CFC-101), antigenicity tests were carried out in guinea pigs and mice. In active systemic anaphylaxis(ASA) test, guinea pigs showed no sign or only moderate sign(1/5) when sensitized and challenged with up to 200 $\mu\textrm{g}$/kg. In homologous passive cutaneous anaphylaxis(PCA) test using guinea pigs, inoculation of CFC-101 alone did not produce CFC-101-specific antibody. When inoculated with 200 $\mu\textrm{g}$/kg plus adjuvant, challenge of 200 $\mu\textrm{g}$/kg produced PCA titer of 32(5/5) but challenge of 20 $\mu\textrm{g}$/kg did not produce CFC-101-specific antibody. In heterologous PCA test using mice, CFC-101-specific antibody was not detected when sensitized with CFC-101 alone. Some animals(3/12) showed positive PCA response when inoculated with 200 $\mu\textrm{g}$/kg plus alum. In passive hemagglutination (PHA) test, although no antibody was detected at 20 $\mu\textrm{g}$/kg, inoculation of 200 $\mu\textrm{g}$/kg alone or with alum produced positive response in all animals. This result has already been predicted because CFC-101 is a vaccine developed for the purpose of immunization. From the above results, it can be concluded that there is no adverse antigenic potential up to 10 times clinical dose of 200 $\mu\textrm{g}$/kg.

• Animal cells and systems
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• v.16 no.3
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• pp.215-223
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• 2012

Recently it has been reported that immunization with heat-killed tumor cells (HK vaccine) induces anti-tumor immune responses in mice. To investigate how HKvaccine elicits anti-tumor specific adaptive immunity, we examined the effect of HK vaccination on innate immune cells such as dendritic cells (DCs), which are essential for the generation of adaptive immunity. Upon stimulation with HK vaccine, DCs matured to promote not only the upregulation of co-stimulatory molecules but also secretion of cytokine IL12. Furthermore, HK vaccine-treated DCs migrated more efficiently to draining lymph nodes compared with untreated ones. Taken together, HK vaccine can be useful as an adjuvant to activate DCs for anti-tumor immune responses.

• Korean Journal of Veterinary Service
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• v.41 no.4
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• pp.251-255
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• 2018

Bordetella bronchiseptica and Pasteurella multocida are two main pathogens responsible for atrophic rhinitis (AR), which causes considerable economic losses in swine industry worldwide. Commercial vaccine has been widely used to prevent the damage from AR in Korea. Adverse effects of vaccination at the injection site have been reported, which results in the numerous complaint from farms. However, data on about local reaction at the injection site remains limited. In this study, we compared the local adverse effects of three commercial vaccines following intramuscular injection. The results showed that no gross lesion was founded at the injection sites of all three vaccines. In histopathologic examination, a various level of lesions was identified. Especially, the local reaction of vaccine including saponin as an adjuvant showed the lowest level of histopathological lesions, when compared to those of oil-based and vitamin E-based vaccines. Therefore, this study would provide the information about the extent of local reaction at the injection site and help the farmer to select AR vaccine in order to avoid adverse reaction due to vaccination.

• 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.