• Clinical and Experimental Pediatrics
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• v.51 no.11
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• pp.1185-1190
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• 2008

Purpose : This study was undertaken to evaluate the immunogenicity and reactogenicity of Td booster immunization in early preadolescents of Korea. Methods : Healthy preadolescents, who had been vaccinated with 4 or 5 doses of DTaP vaccines until 6 years old age, were enrolled in this study from August 2006 to April 2007. Diphtheria and tetanus anti-toxoid antibodies in sera were measured by ELISA just before vaccination and 4 weeks after vaccination to evaluate immunogenicity. Local and systemic adverse reactions observed for 4 weeks after vaccination to access reactogenicity. Results : 183 preadolescents were enrolled and mean age was $11.40{\pm}0.51$ years old. All subjects achieved seroprotective diphtheria and tetanus anti-toxoid antibodies (titers ${\geq}0.1IU/mL$) after Td booster vaccination. Among 183 vaccinees, 73.8% showed local adverse reactions and 37.2% systemic adverse reactions. Pain at injection site (66.1%) was the most common local reaction, and the most commonly shown systemic reaction was myalgia (17.5%). The adverse reactions were spontaneously relieved within three days after vaccination. Conclusion : Td vaccine in this study was high immunogenic and showed an acceptable tolerance in Korean preadolescents. Td booster vaccination at 11-12 years old is the most effective method to increase compliance of the vaccination and to decrease the incidence of diphtheria and tetanus.

• Parasites, Hosts and Diseases
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• v.30 no.3
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• pp.201-208
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• 1992

A Theileria sergenti soluble merozoite preparation containing the 29, 34, 35 and 105 KD as the immunodominant polypeptides, was evaluated for efficacy, safety and protectivity in Holstein calves against virulent field tick challenge. The soluble antigens (100 mg/dose) were fortified with either complete or incomplete Freund's adjuvant. Twenty naive calves, aged one month, were subcutaneously inoculated with the preparation and a booster dose was administered 4 weeks later. Twenty additional calves served as controls. Five weeks after the booster dose, vaccinates and uninoculated controls were moved to a pasture, a heavily tick infested area in Cheju-do, Korea. The vaccinates showed negligible change in hematocrit and total RBC count whereas control animals showed significant (p<0.05) hematological changes and associated anemia. Only 30% of vaccinates required chemotherapy after the experiment was terminated. All control animals required chemotherapy and 25% received blood transfusion. The highest percent parasitized erythrocytes in vaccinated cattle was 0.4% as compared with 3.6% among controls during the month of July. A significant difference (p<0.05) was observed in the rate of body weight increase. Significant difFerences were also noted in serum albumin, aspartate aminotransferase, total protein and bilirubin. Significantly more vaccinated cattle maintained normal ranges of hematological and biochemical values as compared with the control group. It is suggested that soluble merozoite T. sergenti antigens may be potential vaccine candidates for developing a genetic vaccine in Korea.

• Journal of Veterinary Clinics
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• v.21 no.2
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• pp.87-92
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• 2004

This study was undertaken to compare the serological response of dogs to four commercially available combination vaccines and three different vaccination schedules to canine distemper virus (CDV). A total of 120 healthy puppies (20 puppies per group) at 6 weeks of age were randomly assigned to one of four vaccines [C, G, K, and V (or V3) groups] and one of vaccination schedules [V2 and V4 groups]. At six, nine, and 12 weeks of age, puppies in each group were vaccinated with one of four combination vaccines subcutaneously. And puppies in V2 and V4 groups were vaccinated with V vaccine every 2 weeks and 4 weeks, respectively. The serological responses to CDV component of the vaccines were determined by measuring SN titers. The immunogenicity of V vaccine was superior to the other vaccines and optimum vaccination schedule was 3 times vaccination with 3 weeks-interval starting vaccination at 6 weeks of age. Although puppies were vaccinated at 6 weeks of age, the geometric mean CDV titers of puppies in all groups by 9 weeks of age were under the protective level. Therefore, prophylactic measures should include isolation of young dogs from the dog population until vaccination can be expected to provide protection.

• Parasites, Hosts and Diseases
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• v.52 no.1
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• pp.93-97
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• 2014

Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl ${\beta}$-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.

• Korean Journal of Veterinary Research
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• v.25 no.1
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• pp.7-17
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• 1985

Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

• Clinical and Experimental Pediatrics
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• v.50 no.5
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• pp.449-456
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• 2007

Purpose : Antibody persistence after primary series of Haemophilus influenzae type b (Hib) vaccine and responses to a boosters are little known in Korean children. We performed this study to evaluate the antibody titer in relation with a booster immunization of Hib vaccine in Korean children. Methods : One hundred forty-four children aged 12-23 months old were enrolled in three university hospitals. The immunogenicity of a boosters with Hib vaccine was assessed in children previously primed with Hib vaccine. Antibody persistence was also assessed in children who had received 3 doses of Hib vaccine without a booster. Anti-polyribosylribitol phosphate (PRP) IgG antibody levels and bactericidal titers were determined by enzyme immunoassay and bactericidal assay at the Center for Vaccine Evaluation and Study, Medical Research Institute, Ewha Womans University. Results : Prior to a booster in the second year of life, geometric mean antibody concentrations were $2.39{\mu}g/mL$ and the percent of subjects who had a anti-PRP antibody level ${\geq}1{\mu}g/mL$ was 68.6%. After boosting, antibody concentration was $19.09{\mu}g/mL$ and the percent of subjects who had a anti-PRP antibody level ${\geq}1{\mu}g/mL$ was 96.5%, which reflects previous immune priming. In subjects who had finished primary immunization only, the bactericidal titer was 3,946 and in subjects who had a booster, it was 11,205. Anti-PRP antibody level was correlated with serum bactericidal titer. Conclusion : Many children aged 12-23 month old still had protective antibodies after recommended primary immunization only. A booster dose seemed to induce good anamnestic antibody responses in Korean children.

• Pediatric Infection and Vaccine
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• v.8 no.2
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• pp.150-159
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• 2001

Purpose : We need to reconsider booster vaccination schedule of Japanese encephalitis vaccination. To do that we evaluate the long-term immunogenicity and the incidence of adverse events with inactivated mouse brain derived Nakayama Japanese encephalitis vaccine. Methods : We tested neutalizing antibody for 311 elementary school students by plaque reduction neutralizing test(PRNT) at USAMC-AFRIMS(United States Armed Forces Research Institute of Medical Science/Department of Virology). We evaluated vaccine related adverse events by spontaneous reporting prospectively among 15,487 vaccinees who were vaccinated at public health center and 2,277 elementary school students who were immunized previously by a questionnaire and school health record. Results : According to the time interval from the last booster injection of 311 children, PRNT antibody titers gradually decreased as the interval increased; 239 mIU/mL, 188 mIU/mL, 134 mIU/mL, 49 mIU/mL each at 6, 18, 30, 42 months after the last booster injection. The seropositivity rates were 98%, 99%, 95.6%, 71.4% each at 6, 18, 30, 42 months after the last booster injection. There were 21(0.13%) cases with systemic reactions among 15,487 vaccinees who had visited the hospital by prospective passive reporting system at public health center. According to the questionnaires and school health records in elementary school students, local induration and pain were 17.4% and 14.8%, respectively. Systemic reactions including fever, vomiting, rash were reported in few cases. Conclusion : Biannual booster vaccination that has been recommended so far should not be necessary. Surveillance for adverse events with inactivated mouse brain derived Nakayama vaccine should be strengthened to better assess the number of cases and reactions associated with immunization.

• IMMUNE NETWORK
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• v.4 no.1
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• pp.31-37
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• 2004

Background: 1-8D gene is a member of human 1-8 interferon inducible gene family and is shown to be overexpressed in fresh colon cancer tissues. Three peptides 1-6, 3-5 and 3-7 derived from 1-8D gene were shown to have immunogenicity against colon cancer. Methods: To study tumor immunotherapy of these peptides we established an adoptive transfer model. $D^{b-/-}{\times}{\beta}2$ microglobulin (${\beta}2m$) null mice transgenic for a chimeric HLA-A2.1/$D^b-{\beta}2m$ single chain (HHD mice) were immunized with irradiated peptide-loaded RMA-S/HHD/B7.1 transfectants. Spleens were removed after last immunization, and splenocytes were re-stimulated in vitro. Lymphocytes from vaccinated HHD mice were transferred together with IL-2 to the tumor bearing nude mice that were challenged S.C. with the HCT/HHD/B7 colon carcinoma cell line that was found to grow in these mice. Results: Peptide 3-5 was found to be highly effective in CTL activity. Adoptively transferred anti-peptide 3-5 cytolytic T lymphocytes caused significant retardation in tumor growth. Conclusion: This study shows that peptide 3-5 can be the most effective candidate for the vaccine of adoptive immunotherapy against colon cancer.

• Korean Journal of Veterinary Service
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• v.21 no.2
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• pp.117-126
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• 1998

This study was performed to investigate the immunogenicity of autogenous E coli vaccines and their preventive effects on diarrhea in suckling piglets. Autogenous E coli live and killed vaccines were made from the E coli strains isolated from piglets showing diarrhea in field. In group I, pregnant sows were administered with live and killed vaccines at 4 and 2 weeks before parturition, respectively, Killed vaccines were administered twice to pregnant sows at 4 and 2 weeks before parturition in group II, and saline instead of autogenous E coli vaccines was administered to pregnant sows in group III for the control. After parturition, antibody titers in colostrum and milk from sows, incidence of diarrhea in suckling piglets, and immunoreactivity in the ileum of piglets from each treatment group were examined. The results were as follows ; 1. Sixty-two strains of E coli were isolated from suckling piglets with diarrhea. Of the strains, K88 pilus and K99 pilus antigens were identified in 6(9.8%) and 4(6.5%), respectively. Molecular weights of K88 and K99 pilus were 27,500 and 18,500 daltons, respectively. 2. Antibody titers in colostrum from sows after parturition were 1 : 512 to 1 : 1,024 in group I, 1.256 to 1.512 in group II, and 1 : 4 to 1 : 16 in group III. 3. The incidences of diarrhea In suckling piglets of group I, II and III were 3.3%, 9.4% and 21.4%, respectively. 4. When the immunoreactivity in the ileum of piglets from each group was examined, the proportion of IgG-immunoreactivity cells in group I or II was higher than that in group III. In conclusion, administration of autogenous E coli vaccines to pregnant sows before parturition can be an effective way to prevent diarrhea in suckling piglets.

• Parasites, Hosts and Diseases
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• v.53 no.6
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• pp.789-793
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• 2015

In order to explore tick proteins as potential targets for further developing vaccine against ticks, the total proteins of unfed female Dermacentor silvarum were screened with anti-D. silvarum serum produced from rabbits. The results of western blot showed that 3 antigenic proteins of about 100, 68, and 52 kDa were detected by polyclonal antibodies, which means that they probably have immunogenicity. Then, unfed female tick proteins were separated by 12% SDS-PAGE, and target proteins (100, 68, and 52 kDa) were cut and analyzed by LC-MS/MS, respectively. The comparative results of peptide sequences showed that they might be vitellogenin (Vg), heat shock protein 60 (Hsp60), and fructose-1, 6-bisphosphate aldolase (FBA), respectively. These data will lay the foundation for the further validation of antigenic proteins to prevent infestation and diseases transmitted by D. silvarum.