• Title/Summary/Keyword: vaccine development

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IDENTIFICATION OF THE AG I/II AND GTFD GENES FROM STREPTOCOCCUS MUTANS GS-5 (연쇄상구균 GS-5의 ag I/II와 gtfD 유전자 클로닝)

  • Jeong, Jin-Woo;Baik, Byeong-Ju;Yang, Yeon-Mi;Seo, Jeong-Ah;Kim, Jae-Gon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.32 no.2
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    • pp.357-369
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    • 2005
  • Streptococci are Gram-positive, facultative anaerobes and have no catalase activities. Among mutans streptococci containing ${\alpha}-type$ hemolytic activity, S. mutans is a causative agent for dental caries. As well as acid production yielding the demineralization of tooth enamel, adherence and colonization of S. mutans to the teeth are also important for its virulence. These early colonization are accomplished by the bacterial fibrillar protein, Antigen I/II (Ag I/II) and glucosyltransferase (GTF). Therefore, Ag I/II and GTF are reasonable targets for the development of vaccine against S. mutans GS-5. The ag I/II and gtfD genes from S. mutans GS-5 were cloned and sequenced. Sequence analyses showed the nucleotides sequence of cloned genes had high homology to the sequences previously reported. The sequence alignment of 280 nucleotides between the cloned Ag I/II and the available sequence of the corresponding S. mutans GS-5 showed the perfect match. Comparing with the sequence of gtfD from S. mutans UA159, the corresponding nucleotide sequence of S. mutans GS-5 showed some mismatches and the mismatches introduced changes in four residues out of 105 amino acids, yielding four missense mutations.

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Amplified fragment length polymorphism fingerprinting analysis of Staphylococcus aureus isolated from bovine mastitis milk (소 유방염 유래 Staphylococcus aureus의 AFLP 지문분석)

  • Kim, Yeon-soo;Kim, Sang-kyun;Hwang, Eui-kyung
    • Korean Journal of Veterinary Research
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    • v.41 no.2
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    • pp.157-165
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    • 2001
  • Amplified fragment length polymorphism(AFLP) technique is based on the polymorphism detection through selective PCR amplification of restriction fragments from digested genomic DNA and thus includes the procedures of the total DNA digestion by endonucleases, ligation of adapters to the ends of the fragments, and following the selective amplification of the restricted DNA fragments. This study were aimed to : (1) determine the genetic variability of S aureus strains, (2) estimate genetic diversity within and among these strains, (3) compare phylogenetic relationships among these strains as genetic markers using AFLP techniques. Genomic DNA was digested with a particular combination of three restriction enzymes with specific recognition sites and the DNA fragments were ligated to restriction specific adapters and amplified using the selective primer combinations. In the S aureus strain, the number of scorable AFLP bands detected per each primer combination varied from 29 to 102, with an average of 61.59 using 27 primer combinations. A total of 1,663 markers were generated, 904 bands of which were polymorphic, showing a 33.48% level of polymorphism with these primer combinations. Among the primer combinations, E02/T02, E02/T03, E04/H02, E02/T01 and E04/H03 primer combinations showed a high level of polymorphism with 0.78, 0.76, 0.74, 0.71 and 0.70, respectively. But T03/H01, E01/T02 and E01/T03 primer combinations showed a low level of polymorphism with 0.38, 0.37 and 0.15, respectively, Therefore, the former primer combinations will be the most effective for AFLP analysis of S aureus. In SA1 sub-types the level of polymorphism of S aureus KCTC 1927 was similar to that of S aureus CU 01(0.825) and higher than those of other strains such as S aureus CU 02 (0.715), S aureus KCTC 2199(0.625), S aureus KCTC 1916(0.607) and S aureus KCTC 1621 (0.553). In SA2 sub-types the level of polymorphism of S aureus CU 07 was similar to that of S aureus CU 08(0.935) and higher than those of both S aureus CU 04(0.883) and S aureus CU 05(0.883) and lower than those of S aureus CU 03(0.583). In SA3 subtypes the level of polymorphism of S aureus CU 11 was similar to that of S aureus CU 12(0.913) and lower than that of S aureus CU 15(0.623). The results proved that AFLP marker analysis of S aureus strain could be used to study the epidemiology of mastitis and in addition, common genotype in geographic region could be useful for the development of an effective vaccine or DNA marker for easy diagnosis of mastitis caused by S aureus infection.

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Development of mixed Th1/Th2-type immune response in mice following immunization with GP63 from Leishmania donovani (내장리슈만편모충 유래 GP63 항원을 마우스에 접종한 후 관찰되는 Th1/Th2-type 복합 면역반응)

  • Shin, Sung-shik
    • Korean Journal of Veterinary Research
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    • v.41 no.2
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    • pp.211-218
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    • 2001
  • The $M_r$ 63,000 glycoprotein (GP63) and lipophosphoglycan (LPG) of Leishmania donovani were evaluated as vaccine candidates against visceral leishmaniasis. Mice were immunized with liposomeencapsulated GP63 and/or LPG that were purified from the soluble extract of L. donovani promastigotes, and were challenged with virulent amastigotes. Mice immunized with GP63/LPG in liposomes plus BCG resulted in a 27.4% reduction of amastigotes in the liver compared to the control group (liposomes plus BCG), and mice immunized with liposome-GP63 plus BCG failed to induce a protective immune response against the challenge infection. Immunization of mice with GP63 fused to the Schistosoma japonicum glutathione S-transferase (GP63-GST) plus BCG also failed to elicit protective immunity. To analyze the cause of failure to induce protection, cytokine release from the spleen cells of immunized mice and Leishmania-specific serum antibodies were analyzed. Spleen cells from mice immunized with GP63-GST plus BCG that were exposed to soluble extract of L. donovani in vitro produced 10-fold greater quantities of IFN-gamma and 3-fold greater quantities of IL-5 than cells from mice receiving BCG only or saline. Western blot analysis revealed that sera from these mice had Leishmania-specific antibodies recognizing 1 to 3 antigens of L. donovani with M. W. of 60-65 kDa. Although immunization of mice with GP63-GST induced a strong Th1 response, this study indicated that GP63-GST simultaneously elicited the Th2 response of the CD4+ T-cell, which was known to abrogate the protective immune response conferred by the Th1 effector function.

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Analysis of Influenza Virus Isolates in Seoul during 2003-2004 Season (2003-2004 절기 서울지역의 인플루엔자 바이러스 분리 및 아형 분석)

  • Hwang Young-Ok;Lee Jae-In;Seo Byung-tae
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.53-59
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    • 2005
  • Influenza is an important public health problem which occurs almost every winter in temperate climates and is often associated with increased rates of hospitalization and death. In 1999, our influenza surveillance was initiated with 4 voluntary sentinel physicians and the Public Health Center. During the 2003-2004 influenza season, 124 influenza viruses were isolated from 401 clinical specimens, which were collected from patients with Influenza-like illness(ILI) in Seoul. The case definition of ILI is a case with fever more than $38^{\circ}C$ and systemic symptoms; cough, or sore throat. ILI was the highest at the 20-49 age $group(23\%)$ and the rate of virus isolation was the highest at the 7-19 age $group(50\%)$. Among 124 influenza viruses, isolates 83 were identified as A/H3N2 type and others were subtyped as influenza B viruses in 2003-2004 season. Influenza viruses were collected $39.1\%$ at Nowon-Gu, $13.5\%$ Gangnam-Gu and Seocho-Gu etc. and the isolate rate of virus had the area difference; Yongsan-Gu $66.7\%$, Gangnam-gu $50.0\%$, Nowon-Gu $39.9\%$, Kangbuk-Gu $36.8\%$, Seocho-Gu $27.8\%$, Dongjak-Gu $21.2\%$. Out of 401 individuals, 160 was vaccinated $(40\%)$ and the vaccination rate was the highest at the 20-49 age $group(32\%)$. These findings may contribute to the recommondation of the influenza vaccine formulation and the development of influenza control measure.

IFN-${\gamma}$mRNA Expression in Tuberculous Pleural Lymphocytes After in vitro Stimulation with M. tuberculosis Antigens (결핵균 항원 자극에 의한 결핵성 흉수 림프구의 IFN-${\gamma}$ mRNA 발현)

  • Park, Jae Seuk;Kim, Youn Seup;Jee, Young Koo;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.57 no.1
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    • pp.25-31
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    • 2004
  • Background : IFN-${\gamma}$ is the main effector mediator of the host immune response against Mycobacterium tuberculosis. Evaluating the IFN-${\gamma}$ gene expression in response to M. tuberculosis antigens may help in elucidating the host defense mechanism against M. tuberculosis and in the development of a vaccine. Methods : The IFN-${\gamma}$ mRNA expression in the lymphocytes obtained from pleural effusions from tuberculous pleurisy patients (TB-PLC) after in vitro stimulation with whole cell M. tuberculosis(H37Rv), purified protein derivatives(PPD), man-lipoarabinamman (man-LAM), ara-LAM and Antigen 85B(Ag85B) were evaluated. The degree of IFN-${\gamma}$ mRNA expression was determined by a semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) method. Results : M. tuberculosis induced the expression of IFN-${\gamma}$ mRNA in the TB-PLC in time and dose dependent manners. The PPD and Ag85B induced high levels of IFN-${\gamma}$ mRNA expression in the TB-PLC. However, man-LAM inhibited IFN-${\gamma}$ mRNA expression in the TB-PLC, while ara-LAM did not. Conclusion : IFN-${\gamma}$ mRNA expression in TB-PLC is stimulated by PPD and Ag85B, but inhibited by man-LAM.

Coronary Arterial Lesions of Kawasaki Disease Observed in a Mouse Model of Sepsis: A Pilot Study and a Review of the Literature (패혈증 마우스 모델에서 가와사키병의 관상동맥 병변 관찰: 예비연구와 문헌고찰)

  • Kim, Joo-Hyun;Kim, Hyo-Jin;Shin, Jung-Ha;Choi, Ui-Yoon;Lee, Soo-Young;Han, Ji-Whan
    • Pediatric Infection and Vaccine
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    • v.24 no.2
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    • pp.102-107
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    • 2017
  • Purpose: Coronary arterial lesions (CALs) were reported to have developed in children with systemic inflammatory diseases, as well as those with Kawasaki disease (KD). The purpose of this study was to confirm that the CAL development in children with KD occurs in a mouse model of sepsis presenting typical systemic inflammatory response syndrome (SIRS). Methods: To induce the sepsis mouse model with SIRS, 6-week-old C57BL/6 mice were intraperitoneally injected with endotoxin. We compared histological findings of the major organs between the control and the sepsis groups and examined CAL in the heart of the septic mice. Results: Infiltrating inflammatory cells were relatively increased in the heart, liver, and kidneys of the sepsis group, compared with those of the control group. We confirmed lymphocytic infiltration in the myocardium (myocarditis) and the pericardial soft tissue of the heart. Furthermore, coronary artery of the septic mouse was identified, but CAL was not observed. Conclusions: In this study, we failed to confirm the existence of CAL in a mouse model of sepsis. However, it is well-known that CALs are seen in many kinds of diseases that cause SIRS. Our findings suggest further investigation into the clinical significance of CAL in various systemic inflammatory diseases, including KD.

Knowledge based Genetic Algorithm for the Prediction of Peptides binding to HLA alleles common in Koreans (지식기반 유전자알고리즘을 이용한 한국인 빈발 HLA 대립유전자에 대한 결합 펩타이드 예측)

  • Cho, Yeon-Jin;Oh, Heung-Bum;Kim, Hyeon-Cheol
    • Journal of Internet Computing and Services
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    • v.13 no.4
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    • pp.45-52
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    • 2012
  • T cells induce immune responses and thereby eliminate infected micro-organisms when peptides from the microbial proteins are bound to HLAs in the host cell surfaces, It is known that the more stable the binding of peptide to HLA is, the stronger the T cell response gets to remove more effectively the source of infection. Accordingly, if peptides (HLA binder) which can be bound stably to a certain HLA are found, those peptieds are utilized to the development of peptide vaccine to prevent infectious diseases or even to cancer. However, HLA is highly polymorphic so that HLA has a large number of alleles with some frequencies even in one population. Therefore, it is very inefficient to find the peptides stably bound to a number of HLAs by testing random possible peptides for all the various alleles frequent in the population. In order to solve this problem, computational methods have recently been developed to predict peptides which are stably bound to a certain HLA. These methods could markedly decrease the number of candidate peptides to be examined by biological experiments. Accordingly, this paper not only introduces a method of machine learning to predict peptides binding to an HLA, but also suggests a new prediction model so called 'knowledge-based genetic algorithm' that has never been tried for HLA binding peptide prediction. Although based on genetic algorithm (GA). it showed more enhanced performance than GA by incorporating expert knowledge in the process of the algorithm. Furthermore, it could extract rules predicting the binding peptide of the HLA alleles common in Koreans.

Characteristics of carbon dioxide separation using amine functionalized carbon (아민기 개질 탄소를 이용한 이산화탄소 분리 특성)

  • Cha, Wang Seog;Lim, Byeong Jun;Kim, Jun Su;Lee, Sung Youn;Park, Tae Jun;Jang, Hyun Tae
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.4
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    • pp.17-24
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    • 2021
  • The development of a new sorbent for carbon dioxide depends on several factors, such as fast adsorption/absorption velocity, hydrophobicity, and lower regeneration temperature than commercial sorbent. In this study, aminosilane grafted activated carbon was synthesized to capture CO2. Methyltrimethoxysilane (MTMS) and 3-aminopropyl-triethoxysilane (APTES) were used as the grafting precursor of the amine functional group. The APTES grafting activated carbon showed higher sorption property than MTMS used one. The characteristics of the separation mechanism of carbon dioxide were examined by measuring the adsorption capacity according to temperature and carbon dioxide partial pressure. The absorption capacity of carbon dioxide was similar to amine grafting activated carbon and activated carbon at 25℃, but amine-grafted activated carbon was higher at 75℃. The amine functional group-grafted activated carbon showed higher absorption capacity than activated carbon with a 1% carbon dioxide partial pressure. Aminosilane grafting of activated carbon was chemically absorbed but also showed the characteristics of physical adsorption. The reforming activated carbon with an amine functional group grafted solid absorption/adsorption sorbent would significantly impact the material engineering industry and carbon dioxide adsorption process. The functionalized sorbent is a high-performance composite material. The developed sorbent may have applications in other industrial processes of absorption/adsorption and separation.

Epidural Abscess Caused by Eikenella corrodens in a Previously Healthy Child

  • Kim, Ye Kyung;Han, Mi Seon;Yang, Song I;Yun, Ki Wook;Han, Doo Hee;Kim, Jae Yoon;Choi, Eun Hwa
    • Pediatric Infection and Vaccine
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    • v.26 no.2
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    • pp.112-117
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    • 2019
  • Eikenella corrodens rarely causes invasive head and neck infections in immunocompetent children. We report a case of epidural abscess caused by E. corrodens in a previously healthy 13-year-old boy who presented with fever, headache, and vomiting. On physical examination upon admission, there was no neck stiffness, but discharge from the right ear was observed. Brain magnetic resonance imaging (MRI) revealed approximately 4.5-cm-sized epidural empyema on the right temporal lobe as well as bilateral ethmoid and sphenoid sinusitis, right mastoiditis, and right otitis media. During treatment with vancomycin and cefotaxime, purulent ear discharge aggravated, and on follow-up brain MRI, the empyema size increased to $5.6{\times}3.4cm$ with interval development of an abscess at the right sphenoid sinus. Burr hole trephination was performed, and foul-smelling pus was aspirated from the epidural abscess near the right temporal lobe. Pus culture yielded E. corrodens. Endoscopic sphenoidotomy was also performed with massive pus drainage, and the same organism was grown. The patient was treated with intravenous cefotaxime for 3 weeks and recovered well with no other complications. Therefore, E. corrodens can cause serious complications in children with untreated sinusitis.

A Case of Cytomegalovirus Retinitis during Maintenance Chemotherapy for Acute Leukemia

  • Ahn, Bin;Song, Seungha;Han, Mi Seon;Oh, Baek Lok;Choi, Jae Hong;Choi, Eun Hwa
    • Pediatric Infection and Vaccine
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    • v.27 no.3
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    • pp.198-204
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    • 2020
  • Cytomegalovirus (CMV) disease is rare in children who receive anticancer chemotherapy and have no history of stem cell transplantation (SCT). We report a case of CMV retinitis that developed during maintenance chemotherapy for acute leukemia. A 7-year-old boy developed decreased visual acuity and persistent pancytopenia during maintenance chemotherapy. Laboratory investigations initially showed significant CMV antigenemia (51 positive cells/200,000 leukocytes); however, antiviral therapy was not deemed necessary in this patient who had no history of SCT. CMV antigenemia worsened to 170 positive cells/200,000 leukocytes over 3 weeks. Ophthalmological examination revealed multiple bilateral retinal infiltrates and granular lesions. He was diagnosed with CMV retinitis and was treated with a 4-week course of intravenous ganciclovir and intravitreal injection of ganciclovir 6 times, followed by a 1-month course of orally administered valganciclovir. A CMV antigenemia assay showed negative results, and follow-up fundoscopy revealed lesser retinal infiltration after the sixth intravitreal ganciclovir injection. Future studies should focus on the development of standardized screening methods and preemptive therapeutic strategies for CMV disease in high-risk children.