• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.530-530
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• 2000

The host defense system or immune system of all modern animals has their roots in very ancient organisms. After analyzing literature data concerning properties of invertebrates and vertebrates lectins we suggest that mechanism of mannans recognition may exist in marine invertebrates, as a universal mechanism for homeostasis maintenance and host defense, and mannan-binding lectins family of vertebrates has ancient precursor, as was shown for another S-type lectins family. We carried out the screening of mannan-binding type lectin among different species of echinoderms inhabiting in Piter the Grate Bay, the sea of Japan. As a result, the C-type lectins (SJL-32) specific for high mannose glycans was isolated from the coelomic plasma of the sea cucumbers Stichopus japonicus by ion-exchange chromatography on a DEAE-Toyopearl 650M, affinity chromatography on a mannan-Sepharose 6B and gel filtration on a Sephacryl S-200. SJL-32 is homodimer with molecular mass about 32 kDa on SDS-PAGE under non-reducing conditions. Protein part of the lectin has high conteins Asn, Glu, Ser. Hemagglutination of trypsin-treated O blood group human erythrocytes by SJL-32 was competitively inhibited by high-branched -D-mannan composed of -1,2 and -1,6 linked D-mannopyranose residues. In contrast, a variety of mono-, oligo-, and polysaccharides composed of residues of galactose and fucose showed absence or little inhibitory activities. The lectin activity strong depends on Ca2+ concentration, temperature and pH. Monospecific polyclonal antibodies were obtained to the lectin. As was shown by ELISA assay, antibodies to SJL-32 cross-reacted with human serum mannan-binding lectin. This data allows making conclusion about common antigenic determinants and structural homology of both lectins. In our opinion, SJL-32 belongs to evolutionary high conservative mannan-binding lectins (MBLs) family and takes part in the host defense against pathogenic microorganisms.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.215-224
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• 2006

In order to search the availability of the lectin extracted from Korean mistletoe(Viscum album coloratum) as an adjuvant for the avian vaccines, attempts were made to determine toxicity of the lectin in chicks and its immunostimulating activity on the inactivated vaccines against Newcastle disease virus(NDV). For the determination of toxicity, the lectin was injected into the thigh muscle of SPF chicks(Charles River) of 1-week-old and observed hematologically and pathologically. For the determination of immunostimulating effects, lectin-adjuvanted, inactivated NDV vaccines were injected into the thigh muscle of SPF chicks in the same age group. Sera of the chicks were examined for the hemagglutination-inhibition(HI) antibodies induced, their HI titers and reaction to the NDV antigens. The data were further compared with those from aluminum hydroxide [$Al(OH)_3$]-adjuvanted vaccines and vaccines without adjuvant, and the results are as follows. There were no significant changes observed in the values of RBC, WBC, Hb, PCV, MCV, MCH, MCHC, AST, ALT, BUN, creatinine and total proteins in the chicks administered with lectin of 1.1, 2.2 and $22.2{\mu}g/kg$ body weight, which means the lectin has no effects on blood values and functions of liver and kidney. In histopathologic observation, no lesions were observed in the brain, heart, liver, lung, spleen, kidney, thymus and bursa of Fabricius of the chicks administered with lectin of 1.1, 2.2 and $22.2{\mu}g/kg$ body weight. There were inflammatory lesions, such as congestion, hemorrhage, edema, infiltration of macrophages and coagulation necrosis observed in the thigh muscle of chicks administered with lectin of $22.2{\mu}g/kg$ body weight, whereas no changes were observed in 1.1 and $22.2{\mu}g/kg$ lectin administered chicks. In chicks immunized with lectin($4.4{\mu}g/kg$ of body weight)-adjuvanted B1, LaSota and Ulster 2C vaccines, HI titers in reciprocal values for $log_2$ were 1.8-2.2 at 1 week after vaccination, which was similar with those of 1.5-2.9 by $Al(OH)_3$-adjuvanted vaccines. The HI titers by the lectin-adjuvanted vaccines reached to 3.9-5.3 at 4 weeks, whereas those by the $Al(OH)_3$-adjuvanted vaccines were more high as 7.3-9.3. Meanwhile, the immunostimulating effects of the lectin were recognized while compared to the HI titers with 2.4-3.7 in chicks immunized with vaccines without adjuvants at 4 weeks after vaccination. The chicks immunized with lectin-adjuvanted vaccines were enough to resist challenges by Kyojeongwon strain, a very virulent NDV at 4 weeks after vaccination as well as chicks immunized with $Al(OH)_3$-adjuvanted vaccines. The HI titers by the lectin-adjuvanted vaccines reached to high level as 8.7-10.3 as those with 8.2-9.6 by the $Al(OH)_3$-adjuvanted vaccines at 6 weeks after vaccination, which may be the booster effects by the challenge virus. Antibodies specific to the HN and F antigens of NDV were observed in the sera of both chicks immunized with lectin-adjuvanted vaccines and $Al(OH)_3$-adjuvanted vaccines.

• IMMUNE NETWORK
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• v.1 no.2
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• pp.170-178
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• 2001

Background: Korean mistletoe (Viscum album) extract has been found to posses immunostimulatory activity. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to know whether this extract might activate mouse peritoneal macrophages to produce interleukin $1{\beta}$ (IL-$1{\beta}$). Methods: Hemagglutination assay was carried out to examine whether M11C contained a lectin or not. To know the effect of M11C on the production of IL-$1{\beta}$, the macrophages were treated by the M11C, and then collected the supernatant (M11C stimulated macrophages-conditioned media; MMCM). MMCM was analyzed for the IL-$1{\beta}$ quantification and mRNA expression by means of ELISA and RT-PCR, respectively. Results: Maximum effective dose and time of M11C on IL-$1{\beta}$ production from macrophages were $20{\mu}g/m{\ell}$ and 8 hours, respectively. This ELISA data was reconfirmed by immunoblotting assay. indicating that M11C is a good candidate for an immunomodulator. The dose and time dependent effects of M11C on the expression of IL-$1{\beta}$ mRNA from macrophages was also shown in expression of mRNA detected by RT-PCR. Treatment dose and time for the maximum expression of IL-$1{\beta}$ mRNA were $20{\mu}g/m{\ell}$ and 4 hours, respectively. Maximum gene expression of IL-$1{\beta}$ was much earlier than maximum production of it. Conclusion: As results, Korean mistletoe extract, M11C, may be used for an immunomodulator. This will be able to make up for and solve the problems caused by existent immunoagent with many adverse effects through many other studies in future including one molecule extraction.

• Journal of Life Science
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• v.30 no.3
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• pp.221-229
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• 2020

Distylium racemosum is an evergreen tree growing wild on Jeju Island, which has been reported to exert biological activity. Obesity is induced by genetic, metabolic, environmental, and other factors. Among these, certain bacterial and viral infections have been shown to cause obesity, which is known as infectobesity. Human adenovirus (HAdV)-36 is one of the viruses that are known to cause infectobesity in humans. Unlike extensive research on preventing obesity and developing anti-obesity drugs, little research has been conducted specifically on the prevention and treatment of infectobesity. Therefore, this study aimed to evaluate the effects of phytochemicals from D. racemosum on the replication of HAdV-36. A549 cells infected with HAdV-36 were treated with an ethyl acetate fraction of a D. racemosum leaf extract (DRE), and the virus titer was calculated based on the hemagglutination (HA) titer. The results showed a concentration-dependent inhibitory effect of DRE treatment on HA titers. DRE treatment was also found to inhibit the cytopathic effects of the virus and the expression of viral genes. Quercitrin was identified as the constituent of DRE exerting an inhibitory effect on HAdV-36 replication. This study shows that DRE can be used as a candidate substance for the development of treatment for HAdV-36 infections. In addition, this study provides a basis to further investigate DRE for the development of anti-infectobesity medication.

• Journal of Environmental Health Sciences
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• v.42 no.4
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• pp.266-273
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• 2016

Objectives: This study evaluated the virucidal efficacy against avian influenza virus (AIV) of a disinfectant spray containing 0.25% grapefruit seed extract, 0.2% citric acid, 0.0625% malic acid and 0.0125% benzalkonium chloride. Methods: The disinfectant spray was diluted several times with hard water (HW) and organic matter (OM). Two point five mL of each diluent was added into each test tube, and 2.5 mL of AIV suspension was inserted into each test tube. After 30 minutes of virus-disinfectant contact reaction at $4^{\circ}C$, 2.5 mL of 10% inactivated fetal bovine serum was added into each test tube to neutralize the sanitizer efficacy. The neutralized solutions were serial 10-fold dilutions with phosphate buffer solution, and 0.2 mL of the diluents was injected into the allantoic cavity of five ten-day-old-chickens per dilution time. After incubation of the embryos for five days, the viability of the AIV was examined by hemagglutination titer. The valid dilution of the disinfectant spray was estimated according to the dilution time that the virus titer was inactivated more than $10^4$ 50% egg-infective dose (EID50)/mL compared with pathogen control. Results: In HW and OM conditions, the valid dilutions of the disinfectant spray against AIV were seven- and three-fold dilutions, respectively. The AIV titer of the pathogen control was more than 6.1 log10EID50/mL, and there was no embryonic toxicity. Conclusion: The present study showed that this disinfectant spray has effective virucidal activity against AIV.

• 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.

• Korean Journal of Poultry Science
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• v.28 no.2
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• pp.83-89
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• 2001

Newcastle disease virus, CBP-1 strain isolated from Korean pheasants was passaged for 173 times by 9-day-old specific pathogenic free (SPF) embryonated eggs at $37^{\circ}C$ (parent strain) and subsequently passaged for 15 (cold attenuation (CA) -15) and 30(cold attenuation (CA) -30) times by 10-day-old of commercial broiler chicks embryonated eggs at $29^{\circ}C$, respectively, The Physical and chemical properties (sensitivity to lipid solvents, low pH and thermostability), pathogenicity (mean death time, intracerebral pathogenic index and intravenous patho-genic index), safety, booster or protective effect and characterization of temperature sensitivity were measured in cold attenuated CA-15 or 30 strain and compared to those of parent CBP-1 strain. NDV, CBP-1 CA-30 strain acquired cold attenuation and decreased infectivity at $41^{\circ}C$ compared to those of parent strain grown at $37^{\circ}C$. It lost hemagglutination activity (HA) and cell infectivity at $56^{\circ}C$ for 30, 60, and 120 Min. CA-30 strain treated with ethyl ether also lost its HA and cell infectivity. Both CA-30 and parent strains exhibited a little resistant to HA at pH 3.0 glycine HCI buffer. Intracerebral pathogenic index (ICPI) and intravenous pathogenic index (IVPI) of parent strain were 1.12 and 1.45, but decreased to 0.75 and 0.00 in CA-30 strain, respectively. The safety was evaluated by mortality in chicks inoculated with 10$^{4.0}$ EID$_{50}$ /0.1 ml. The mortalities of parent, CA-30 and commercial Bl strains were 17.5, 12.0 and 0.0%, respectively. The safety of CA-30 strain was higher than that of parent strain. The booster effects of CA-30 strain and parent strain performed in 4-week-old chicks after being vaccinated with primary commercial Bl strain.