• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.946-950
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• 2001

Slow-growing pathogenic mycobacterium species, including Mycobacterium bovis BCG, secrete a large amount of glutamine synthetase into culture media. Extracellular and intracellular glutamine synthetases were purified from M. bovis BCG. While the native molecular weights of both glutamine synthetases were estimated to be 370.2 kDa, those of the subunits were 61.7 kDa, indicating that the native forms were composed of 6 subunits. The enzymes showed a hhigh thermal stability and high degree of sequence similarity with the glutamine synthetase from M. tuberculosis in the N-terminal amino acid sequence. Western blotting analysis indicated that the antibodies prepared against both the extracellular and intracellular enzymes exhibited common antigen determinants.

• 대한수의학회지
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• 제32권1호
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• pp.117-125
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• 1992

Mycobacterium (M) bovis 를 인공감염시킨 개에 있어서 BCG의 전처치효과를 in vivo 및 in vitro에서 검토하였다. 개들은 BCG 전처치군, M bovis 단독처치군, 비감염대조군의 세군으로 나누었다. BCG는 M bovis 복강접종 3주일전에 0.2ml를 피내접종하였다. 결핵균 투여 4개월후에 전군을 도살하여 실험에 사용하였다. 도살시 모든 처치군에서 감염이 확인되었다. 병리조직학적으로 BCG전처치군의 폐장내에서는 경도의 macrophage의 침윤과 소상의 육아종 형성이 관찰되였으나 M bovis 단독처치군에 있어서는 보다 고도의 macrophage의 침윤, 중등도의 호중구의 침윤 및 중등도의 육아종의 형성이 확인되었다. 각 동물의 기관지폐포세정액을 분리하여 그 속의 총세포수와 각 세포의 분획을 검토하였다. 비감염 대조군의 기관지폐포세정액내의 총세포수는 두 처치군보다 훨씬 낮았으며 M bovis 단독처치군의 총세포수는 BCG 전처치군보다 1.8배 높았다. 이 세정액으로부터 폐포 macrophage를 분리배양하여 macrophage의 활성능과 결핵균의 증식능을 관찰하였다. BCG처치군은 M bovis 단독처치군에 비하여 높은 Fc receptor 활성(rosette 형성능, 탐식능)과 낮은 결핵균의 증식이 관찰되었다. 그러나 BCG의 전처치는 결핵균을 killing하지는 못하였다. 개에게 BCG를 전처치하면 폐내에 극소수의 결핵균이 지속적으로 잔존하지만 폐포 macrophage는 이미 항결핵성면역능을 지닌채로 계속 활성화된 상태로 존재하기 때문에 결핵에 대하여 예방효과를 갖는다고 사료된다.

• Bulletin of the Korean Chemical Society
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• 제35권3호
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• pp.933-937
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• 2014

A derivative of Mycobacterium bovis Bacillus Calmette-Guerin (BCG) has been used for the preparation of tuberculosis vaccines. To establish a Korean tuberculosis vaccine derived from BCG-Pasteur $1173P_2$, genome sequencing of a BCG-Korea strain was completed by Joung and coworkers. A comparison analysis of the genome sequences of the BCG-Pasteur $1173P_2$ and BCG-Korea strains showed marginal increases in the total genome length (~0.05%) and the number of genes (~4%) in the BCG-Korea genome. However, how the genomic changes affect the BCG-Korea protein expression levels remains unknown. Here, we provide evidence of the proteomic alterations in the BCG-Korea strain by using a SWATH-based mass spectrometric approach (Sequential Window Acquisition of all THeoretical mass spectra). Twenty BCG proteins were selected by top-rank identification in the BCG proteome analysis and the proteins were quantified by the SWATH method. Thirteen of 20 proteins showing significant changes were enough to discriminate between the two BCG proteomes. The SWATH method is very straightforward and provides a promising approach owing to its strong reliability and reproducibility during the proteomic analysis.

• Pediatric Infection and Vaccine
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• 제18권1호
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• pp.91-96
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• 2011

BCG는 M. bovis로부터 유래된 생백신으로 5-6세 미만의 소아 결핵 특히 결핵성 수막염이나 속립성 결핵과 같은 중증 결핵의 예방 효과가 인정되고 있다. BCG 접종 후 이상반응은 국소 궤양 형성, 국소 림프절염이 가장 흔하며, 드물지만 골수염, 파종성 감염과 같은 치명적인 합병증도 있다. 파종성 BCG 감염은 주로 면역저하 환자에게서 흔하게 관찰되며 면역 저하가 없는 소아에게서 발생한 파종성 BCG 감염은 국내 문헌상 보고가 없었다. 저자들은 13개월 여환이 BCG 접종 5개월 뒤 발생한 발열, 피부 결절 및 다발성 림프절 종대를 주소로 내원하여, 백 신 접종 부위 외에 2군데에서 M. bovis BCG를 확인한 후 파종성 BCG 감염으로 진단된 1례를 경험하였다. 환아는 isoniazid와 rifampin으로 치료받았으며 약 2주 간의 입원 치료 후 임상 증상이 호전되어 퇴원하였다. 3주뒤 외래 추적 관찰에서 역설적 반응으로 생각되는 림프 절 종대와 피부 결절이 진행하였으나 동일 약제를 유지하면서도 다시 임상 증상이 호전되었고 환아의 면역학적 검사상 이상 소견이 관찰되지 않았기에 이를 보고하는 바이다.

• Clinical and Experimental Vaccine Research
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• 제13권3호
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• pp.184-201
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• 2024

Tuberculosis (TB) is consistently ranked among the deadliest diseases worldwide, causing millions of deaths annually. Mycobacterium tuberculosis is the causative agent for this infection. Different antibiotics and vaccines have been discussed as potential treatments and prevention. Currently, there is only one licensed vaccine against TB, Bacillus Calmette-Guérin (BCG). Despite its protective efficacy against TB in children, BCG has failed to protect adults against pulmonary TB, lacks therapeutic value, and can cause complications in immunocompromised individuals. In this review, BCG, the most widely administered vaccine, is discussed, and the newest vaccines available in medicine are discussed. Based on the restrictions that prevent optimal BCG efficacy and the vaccines that are now being tested in various clinical studies, some criteria need to be considered in designing future vaccines.

• 생명과학회지
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• 제15권2호
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• pp.182-185
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• 2005

IS1096 transposon을 사용하여 결핵균군에 작용하는 항생제인 PA-824에 내성을 나타내는 Mycobacterium bovis BCG의 돌연변이 균주를 얻고자 하였고, 그 결과 24종류의 서로 다른 돌연변이 균주를 얻을 수 있었다. Transposon이 insertion된 부위를 알아내기 위해서 각각의 돌연변이 균주로 inverse PCR을 수행하였고, PPE 유전자를 포함한 여러 가지 부위에 insertion된 transposon의 위치를 확인할 수 있었다. PPE 유전자에 insertion돌연변이가 발생한 5개 균주의 세포 추출물을 HPLC로 분석한 결과, 3개에서는 야생균주에서 관찰되는 coenzyme $F^{420}$이 존재하지 않았고, 그 생합성 경로의 중간산물인 F0만 존재하였다. 또한 나머지 2개에서는 F0 또는 $F^{420}$어느 것도 존재하지 않았다. 이 결과는 PPE 유전자들의 산물들이 coenzyme $F^{420}$에 어떤 식으로든 관여하고 있음을 나타낸다고 할 수 있다.

• 한국동물위생학회지
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• 제30권2호
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• pp.233-241
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• 2007

Tuberculin positive cattle without gross tubercle lesions should be confirmed by the bacteriological examination to determine the state of the infection. To overcome the time-consuming and laborious identification by culture and biochemical tests, polymerase chain reaction (PCR) has been used to identify Mycobacterium bovis. Due to various lipids in the cell wall of Mycobacterium spp, novel methods of DNA extraction from Mycobacterium spp have been developed. In this study, a newly developed guanidium isothiocyanate/silica DNA extraction method was directly applied to specimens from the tuberculin positive cattle. DNAs were directly extracted from the lymph nodes and the major polymorphic tandem repeat (MPTR) and mycobacterial protein of BCG 70 (MPB70) were amplified using PCR. The DNA extraction method using guanidium isothiocyanate/silica was efficient and safe, and the MPTR and MPB70 primers were specific to M bovis. Therefore, MPTR and MPB70 PCRs will be useful for the detection of M bovis in the lymph node from skin-test positive cattle.

• 대한수의학회지
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• 제36권1호
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• pp.31-38
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• 1996

1. The sixty raised shepherd and sixty-five inhoused pet dogs in the regions of Daejon and Cheongju were subjected to investigate the TB infection by means of BCG and X-ray diagnosis. The 5 out of 65 inhoused pet and 7 out of 60 shepherd dogs were observed to be infected with TB, respectively. However, none of Mycobacterium species were detected from lung tissues of 4-slaughtered dogs showing BCG positive reaction. 2. The rats were first inoculated with 0.1ml BCG, and then 0.1ml M bovis suspended solution($1{\times}10^5$ organisms/0.2ml) 3weeks later. After 5 months, the animals were killed. The pathohistological results from both groups, TB inoculated and BCG treated groups, were observed on the surface of lung. Furthermore, the severe pathological lesion in the Iung was observed in M bovis inoculated group compared to BCG treated group. 3. The slight macrophage invasion and granuloma formation in the lung from BCG treated group were observe individually. However, it was confirmed that the lung from M bovis treated group was invaded by the macrophages and neutrophils combined with the granuloma formation. 4. When the numbers of the total cells taken from broncho-alvealar fluid in each of mouse from both groups were differentially counted, the number of total cell, neutrophils, and lymphocytes from M bovis treated group were significantly increase compared with those of BCG treated group. 5. Although there were nearly no response of the alveolar macrophages to CSF in serum obtained from control group, those from M boris treated group were significantly proliferated.

• BMB Reports
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• 제34권4호
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• pp.342-346
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• 2001

The Mycobacterium bovis BCG panB gene, encoding ketopantoate hydroxymethyltransferase (KPHMT), was cloned from a ${\lambda}gt11$ genomic library and sequenced. The DNA sequence encodes a protein that contains 281 amino acid residues (M, 29,337) with a high similarity to the KPHMTs. Subcloning of a 846 by open reading frame (ORF), but not a 735 by ORF, into the vector pUC19 led to complementation of the panB mutant of Escherichia coli. The BCG pang gene was overexpressed in E. coli and the KPHMT purified to homogeneity The recombinant protein was further confirmed by an enzymatic assay.

• 대한미생물학회지
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• 제35권2호
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• pp.117-127
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• 2000

The non-ELR-containing CXC chemokines Mig and IP-10 have been shown to function as chemotactic cytokines for activated T lymphocytes. In this study, we examined the potential involvement of Mig and IP-10 in antimycobacterial response of mice immunized or infected with M. bovis BCG. The accumulation of Mig and IP-10 mRNA in resident peritoneal monocytes ($RPM{\Phi}$) was slightly reduced by stimulation with vBCG, and the degree was greater for 24 hr culture even though IFN-${\gamma}$ was added. Expression of Mig, IP-10, and IFN-${\gamma}$ in 24 hr delayed-type hypersensitivity (DTH) response was stronger in vBCG-immune mice than in the non-immune. The increase of DTH measured by foot-pad thickness appears to be clearly related to the levels of chemokines Mig and IP10 messages and those of IFN-${\gamma}$ and IL-12. Stimulation with vBCG for 2 days decreased or completely dropped the levels of Mig message in non-immune or immune splenocytes, respectively, whereas IP-10 message was slightly decreased in 2 days culture. Moreover, messages for IL-12 (p40) showed similar kinetics for Mig. The levels of Mig and IP-10 mRNA during the course of infection with BCG were not readily changed in lungs, livers, and spleens from BCG-infected mice. Although there was no obvious changes of Mig and IP-10 messages in the target organs during infection process, we found that the infection progressed over the first 3 wk before being contained by the emerging immune response suggested from detectable amount of IFN-${\gamma}$ mRNA around this time. In view of selectivity of chemokines Mig and IP-10 for activated T cells, these data suggest that chemokine Mig and IP-10, especially in collaboration with IL-12 and IFN-${\gamma}$, may playa role as T cell recruiters in immune response against mycobacterial infection.