• Microbiology and Biotechnology Letters
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• v.23 no.1
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• pp.110-117
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• 1995

This study was carried out to immunologically characterize Bacillus thuringiensis (B.t) antigens. Protein patterns of ultrasonicated- antigens of B. thuringiensis subspecies using SDS- PAGE revealed marked similarities among all the strains analyzed except for the difference between quantative variations of bands and some protein antigens. The comparison of the protein patterns showed that the protein antigen of 45 kilodalton (kd) was common in 11 strains and that the difference between B. thuringiensis subsp. canadensis and galleriae was noticed in quantative variations of bands despite of ambiguous serogrouping, suggesting a useful method for identification. All strains examined showed similar antigenic patterns in SDS-PAGE, while immunodominant bands differed in antigenic reactivity in western blot using polyclonal antibodies. Polyclonal antibody to B. thuringiensis subsp. thuringiensis and israelensis in indirect immunofluorescence assay reacted with flagella and cell surface antigens. The present study indicates that SDS-PAGE and western blot analysis may be used as tools for differentiation and identification of B. thuringiensis subspecies.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.230-235
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• 2005

For the acellular pertussis vaccine with a high immunogenicity, the concentration, composition and characteristics of acellular pertussis antigens are the crucial points to be considered. Nevertheless, it has not been proved yet whether or not the polymerization degree, one of the characteristics of formalin-detoxified acellular pertussis antigens, has an influence on vaccine potency. Thus, in the present study, the correlations among detoxification conditions of acellular pertussis bulks, their polymerization degrees and their immunogenicities were examined. In addition, the relative importance of pertussis toxoid in vaccine immunogenicity was also investigated. Results show that a lower lysine concentration during detoxification induces highly-polymerized antigens, the immunogenicity has a great dependency on the polymerization degree of antigens, and also pertussis toxoid has a relatively stronger influence on the immunogenicity than other antigens. Accordingly, in the aspect of the potency of detoxified acellular pertussis vaccine, it can be demonstrated that the polymerization of antigens and its degree are the major factors affecting the immunogenicity along with a relatively high content of pertussis toxoid

• Korean Journal of Veterinary Research
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• v.32 no.2
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• pp.195-205
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• 1992

The soluble antigen profiles and antigenic specificities of Leptospira interrogans serovars icterhaemorrhagiae, canicola, pomona and hardjo were examined by SDS-polyacrylamide gel electrophoresis, crossed immunoeletrophoresis and immunoblotting. The profiles of protein, glycoprotein and fraction containing N-acetylglucosamine of 4 serovars were compared. The protein profiles of 4 serovars were very similar except the range of 14,400 to 30,000 daltons. Molecular weight of glycoprotein of L, pomona was lower than other serovars. L canicola showed extra N-acetylglucosamine bands having molecular weight of 82,000 and 90,000 daltons. In crossed immunoelectrophoresis, a close antigenic relationship was found between L icterohaemorrhagiae and L canicola. In immunoblottings conducted with soluble antigens and rabbit antisera of 4 serovars, Leptospira interrogans serovars possessed cross-reactive antigens and serovar-specific antigens. The molecular weights of serovar-specific antigens were 45,000, 82,000 and 90,000, 31,000 and 24,000 daltons in L icterohaemorrhagiae, L canicola, L pomona, and L hardjo, respectively.

• The Korean Journal of Zoology
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• v.34 no.1
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• pp.44-53
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• 1991

Distribution of the antigens during the cell cycle of amoebae was followed by immunof-luorescence microscopy using a monoclonal antibody against the nucleus as a probe. While the cells were in the interphase, the antigen was localized on the nucleus membrane. But it was dispersed all over the cytoplasm during mitosis and cytokinesis. The molecular weights of the immunoreacted antigens were 210 KD and 190 KD as determined by SDS PAGE and western blotting of the purified nuclei. The antigens were not soluble in non-ionic detergent, but were released from the nucleus by incubation with 0.05 M sodium carbonate, pH 10.6 or with 8 M urea at serial chemical extraction. Thus the antigens appeared to be peripheral proteins of the nurBeus envelope. The isoelectic point of both antigens was 7.64 as determined by 2 D PAGE and transfer blotting. Considering the peiipherd association with the nucleus membrane and the dispersed distribution during mitosis, the antigens could be lamin like proteins. Hourever, it appears also possible that they are the component molecules of the unusually structured aurous lamina of amoeba nucleus since they have the large molecular weight and the basic pl.

• Korean Journal of Veterinary Research
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• v.44 no.4
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• pp.625-635
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• 2004

Trichnuris suis does not excrete eggs during larval stage as well as in particular adult stage, It is impossible to diagnose by use of fecal examination method in those periods. Therefore, serological diagnostic method can be very useful for those stages. In order to produce monoclonal antibody, specific somatic and secretory-excretory (SE) antigens of T. suis were identified and analyzed by SDS-PAGE and Western blot. Monoclonal antibody-producing hybridoma cells were cloned, which were made of popliteal lymph node of BALB/c mice immunized with a 45 kDa somatic antigen of T. suis. Five clones (1B9, 2C4, n2C5, 2D7 and 2D8) showing strong responses to T. suis antigens were selected and the isotype identified. All monoclonal antibodies were IgG1 isotype and the light chains were k chain. Established monoclonal antibodies reacted specifically to somatic and SE antigens of T. suis and did not cross-reacted to antigens of ascaris suum, trichuris vulpis, or Trichinella spiralis. The sensitivity of somatic and SE antigens against these monoclonal antibodies were significant (p<0.01) associated with those of positive and negative sera.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.829-836
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• 1998

Immunogenicity of Escherichia coli pilus and LT were evaluated in 20-week-old hens. The antigens were consisted of K88, K99, 987p pilus and heat labile toxin purified from enterotoxigenic Escherichia coli. The durations of antibody titers in sera and egg yolk were investigated by an enzyme-linked immunosorbent assay(ELISA). After first inoculation, antibody titers in sera reached at peak 2 weeks postinoculation. However, peak antibody titers in egg yolk were detected 4 weeks postinoculation, indicating that transfer of immunoglobulin from serum to egg yolk took about two weeks period. Although there were slight reduction in titers, the specific antibodies in egg yolk lasted up to 3 months. Immune responses against monovalent and combined antigens were showed as almost same patterns. The transfer rate of antibodies from serum to egg yolk didn't show any significant differences among three pilus antigens in this study. Considering the concentrations of antigens in each inoculated group, multivalent antigens containing heat labile toxin of E coli were found to be more immunogenic than monovalent antigen in producing specific antibodies. From this experiment, it was demonstrated that multivalent antigens containing three pilus and heat labile toxin could be a promising candidate for the production of egg yolk antibodies for prophylactic use in preventing swine colibacillosis in future.

• IMMUNE NETWORK
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• v.7 no.1
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• pp.26-30
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• 2007

Background: Mycobacterial antigens released as PIM, LM, LAM, lipoproteins and other cellular factors may contribute to macrophage and dendritic cell activation through pattern recognition receptors such as TLRs. In this study, we assessed cytokine production and ERK activation with stimulation of several major mycobacterial antigens. Methods: Purified mycobacterial antigens (10, 22, 30, 38kDa) and recombinant antigens (6, 16, 19, 38kDa, Ag85A antigen) were studied. The production of cytokines (TNF-${\alpha}$, IL-12, IL-6) was measured by ELISA. The ERK activation was detected by western blotting. The expression of TLR2 or TLR4 was measured by flow cytometry. Results: Among purified antigens only 30kDa antigen induced production of IL-6 or TNF-${\alpha}$ in THP-1 macrophage cells. When THP-1 macrophage cells were treated with 30kDa antigen, phosphorylation of ERK was detected. ERK activation also occurred in TLR2 transfectant HEK293 cells with 30kDa antigen stimulation. Conclusion: 30kDa antigen is one of the major mycobacterial antigens inducing cytokine production and MAP kinases phosphorylation in macrophages.

• Korean Journal of Veterinary Research
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• v.30 no.3
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• pp.297-307
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• 1990

To establish an agar-gel immunodiffusion (AGID) test for detection of antibodies to Aujeszky's disease virus(ADV) in swine, the precipitating antigens were prepared by four procedures using the Aujeszky's disease virus, NYJ-1-87 strain isolated from the affected piglets in Korea. The optimal condition for AGID test and the properties of the antigens were investigated. To determine the optimal concentration of antigens, four antigens were experimentally prepared by concentrating the viral fluids by 1/30 to 1/200. It was proved that the antigen precipitated with ammonium sulfate at concentration of 1/100 was the most efficient to detect ADV antibodies by AGID test. When the relationship between the concentration of the antigens and the size of precipitating in radial immunodiffusion test was investigated, a high correlation coefficiency at r=0.95 (y=0.23x+23.4) was estimated, In study on the effects of various buffered salt solutions and agars on the sensitivity of AGID test by using the experimental ADV antigens, it was found that 0.05M tris buffer without sodium chloride at pH 7.2 induced the most distinctive precipitating lines, and that there was no significant differences in the sensitivity between the agarose and Noble's special agar. When the efficiency of AGID test was compared with serum neutralization(SN) test, the sensitivity of AGID test was 100% in SN titer over 1 : 16, 91.7% in SN titer of 1 : 8 and 57.1% in SN titer of 1 : 4. The specificity of AGID test compared with the sera with SN titer under 1 : 2 was 98.4%. Protein analysis of the antigens by SDS-PAGE indicated that antigen I and antigen III showed a specific band of polypeptides with molecular weight of 116 K in comparison with the control antigen. Antigen IV, treated with tween-80 and ammonium sulfate, revealed specific polypeptides bands at the molecular weights 45K, 98K and 150 K.

• Korean Journal of Veterinary Research
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• v.43 no.4
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• pp.689-696
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• 2003

Canine sarcoptic mite (Sarcoptes scabiei var. canis) burrow usually in the stratum corneum of the skin of dogs and rabbits. Antigens from the burrowing mites induce cutaneous inflammatory reaction and humoral and cell-mediated immune response in the host. The effect of immunization induced by somatic antigens of house dust mite (Dermatophagoides spp.) has been evaluated to control the canine sarcoptic mite in this experiment. Twelve common antigens (187, 142, 126, 120, 109, 92, 80, 68, 51, 30, 25, 17 kDa) were found using SDS-PAGE with silver staining and Western blot between canine sarcoptic mite and house dust mite. In order to evaluate the immunologic effect of these common antigens 10 New Zealand white rabbits were divided as 4 groups such as negative control (group I), positive challenged control (group II), vaccinated (group III), and vaccinated-challenged (group IV) groups. Group II was artificially infested with about 1,000 canine sarcoptic mites and group III and IV were immunized with somatic antigens of house dust mite. In addition group IV was artificially infested with about 1,000 canine sarcoptic mites and group II, IV were treated with ivermectin. At the 8 weeks of the vaccination with common antigen, the antibody titers of all groups of II, III and IV had been increased. Both infestation score and live canine sarcoptic mite counts of group IV were lower than group III. Infestation score of group II become 0 by 2 weeks and group IV by 4 weeks after infestation. These results suggest that house dust mite, which is easy to culture in vitro, can be a vaccine candidate for protection of canine sarcoptic mite infestation.

• Korean Journal of Veterinary Research
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• v.42 no.2
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• pp.253-260
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• 2002

Fowl red mite, Dermanyssus gallinae, is the most important ectoparasite affecting egg layers worldwide. More than 35 compounds have been used for fowl red mite control. Although some of them are efficient, several compounds are unsuitable in terms of food safety and environmental problems. Some compounds are efficient in theory but inadequate in practice. It is also expensive in material and labor to control effectively. Effective doses are very close to toxic doses and repeated treatment is required. Repeated, long term treatment of compounds on fowl red mite populations, may cause heritable resistance against the mites. In this study, antigenicity of fowl red mite and house dust mite, Dermatophagoides pteronyssinus, were identified by SDS-PAGE, silver staining, Western blotting and ELISA to investigate immune effects against fowl red mite using somatic antigens of fowl red mite and house dust mite. By SDS-PAGE, silver staining and Western blotting, several common antigens (110, 60, 56, 49, 46 kDa) of both fowl red mite and house dust mite were recognized. To identify immune effect of somatic antigens of fowl red mite and house dust mite, sixty white leghorn broilers(1 week old) were used. Among sixty white leghorn broilers, twenty were immunized with fowl red mite somatic antigens(Group I), twenty immunized with house dust mite antigens(Group II), and twenty were control group without antigen(Group III), respectively. After immunization, it was identified that antibody titers were increased both in group and II. Then all groups were challenged with fowl red mites. After 2 months, measurements of body weights, packed cell volume(PCV), ELISA OD values and numbers of mites were significant(p<0.05). These results suggest that fowl red mite and house dust mite, which are easy to collect and maintain, can be good vaccine candidates against fowl red mite in chicken.