• 한국동물학회지
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• 제37권4호
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• pp.531-537
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• 1994

Partial internal amino acid sequences of the 15S ATPase from chick skeletal muscle were determined and found to be identical to the corresponding regions of the Mg2+_ATPase from Xenopus laevis oocytes, that is a close homolog of N-ethvlmaleimide-sensitive factor (called NSF) in hamster and Sec18p in yeast, both of which are believed to plaN an essential role in vesicle fusion in secretory process. Thus, the 15S Arpase in chick skeletal muscle maw also belong to a protein family of the "vesicle fusion proteins". Unlike the Mg2'-Afpase with an isoelectric point (pl) of 5.5, however, the 15S Arpase was separated into four spots with pls of 4.9,6.4 and 6.9 upon analysis by twoiimensional gel electrophoresis. In addition, the anti-15S ATPase IgG was found to be capable of interacting with the 265 protease complex upon analysis by immunoprecipitation. Moreover, immunoblot analysis revealed that the anti-155 Arpase IgG recognizes three subunits, ko of which show the same mobilities as the 510-kDa subunit 4 and 48-kDa subunit 7 of the 26S protease complex that are known to contain a highly consented ATP-binding motif. These results surest that a common antigenic site, likely the consensus nucleotide-binding site, exists in the 15S ATPase and the 26S pretense complex and hence both the enzymes maw also be related ATPases.d ATPases.

• 생명과학회지
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• 제20권3호
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• pp.365-373
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• 2010

2006년 10월부터 2008 년 6월까지 총 인플루엔자 의사 환자 1,822건의 인후도찰물 및 비인후도찰물에서 277건의 인플루엔자바이러스를 분리했다. 절기별로는 2006~2007 절기의 1,154검체 중 52건(4.5%), 2007~2008절기의 668검체 중 210건(31.4%)에서 인플루엔자바이러스를 분리하였다. 인플루엔자바이러스 A/H1N1의 HA 유전자의 경우, 2008~2009 절기의 백신주인 A/Brisbane/59/2007과는 96.7%~97.7%, A/Solomon Islands/3/2006 96.5%~97.3%, A/New Caledonia/20/99와는 95.6%~96.6%의 유사성을 나타냈으며, NA 유전자의 경우, A/Brisbane/59/2007과는 97.8%~98.5%, A/Solomon Islands/3/2006과는 96.7%~97.6%, A/New Caledonia/20/99와는 96.8%~97.7%의 유사성을 보여 2008~2009절기의 백신주인 A/Brisbane/59/07과 가장 유사성이 컸다. 인플루엔자바이러스 A/H3N2의 분리주 중 1주를 제외한 모든 분리주가 HA 유전자에서 2008~2009 절기 백신주인 A/Brisbane/10/2007과는 98.4%~99.7%의 유사성을 보였고, A/Wisconsin/67/2005와는 96.5%~97.5%의 유사성을 보였으며, NA 유전자에서는 A/Brisbane/10/2007과는 98.9%~99.4%, A/Wisconsin/67/2005와는 98.0%~98.6%, A/California/7/2004와는 98.3%~98.9%의 유사성을 보였다. 인플루엔자바이러스 B의 HA 유전자의 경우는 2주를 제외하고는 2008~2009 절기의 백신주인 B/Florida/4/2006과는 96.5%~99.7%의 유사성을 보였으며, B/Malaysia/2506/2004와는 86.7%~87.7%의 유사성을 보여 B/Florida/4/2006과의 유사성이 크게 나타났다. NA 유전자의 경우는 reassortant분리주가 96.7%와 97.3%의 유사성을 나타내는 것을 제외하고는 B/Florida/4/2006에 98.9%~100%의 유사성을 나타냈으며, 분리주 유행시기의 백신주인 B/Malaysia/2506/2004와는 94.5%~96.7%의 유사성을 나타내어 2008~2009 절기의 백신주와 더 큰 유사성을 보였다. HA 유전자에서는 conserverd receptor binding site는 아미노산의 치환 없이 모든 분리주에서 잘 보존되어 있었으며, N-linked glycosylation site도 인플루엔자바이러스 A/H1 1주, A/H3 1주를 제외하고는 모두 같은 수의 N-linked glycosylation sites를 가졌으며, 인플루엔자바이러스 B의 경우는 2008~2009 절기의 백신주보다 1개가 많은 4개의 N-linked glycosylation sites를 가지고 있었다. Antigenic sites의 경우는 인플루엔자바이러스 A/H1의 Sb의 3개의 아미노산에서 백신주들과 다른 아미노산을 가지고 있으며, A/H3에서는 A, B, E 부위에서 는 아미노산의 변화가 나타났고, C, D 부위에서는 변화가 없었다. 인플루엔자바이러스 B의 4개의 분리주에서는 150 loop와 160 loop에서 B/Florida/4/2006과 비교하여 1개의 아미노산에서 치환이 나타났으며, 190 helix에서 모든 분리주가 B/Florida/4/2006과 비교하여 1개의 아미노산에서 치환이 나타났다.

• Animal cells and systems
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• 제3권2호
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• pp.221-228
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• 1999

In order to find a biochemical marker for late Iysosomes, we characterized two cDNAs which were cloned by using a monoclonal antibody (mAb) against Iysosomes in Amoeba proteus as a probe. The two cDNAs, a 1.3-kb cDNA in pBSK-Iys45 and a 1.6-kb cDNA in pBSK-Iys60, were found to encode proteins homologous to pepstatin-insensitive carboxyl proteinases (PICPs). E. coli transformed with pBSK-Iys45 produced two immunopositive polypeptides (45 and 43 kDa) and the cDNA in 1274 bases encoded a 44,733-Da protein (Lys45) of 420 amino acids containing one site for a core oligosaccharide. On the other hand, E. coli transformed with pBSK-Iys60 produced several polypeptides (64, 54, 45, 41, and 37 kDa) reacting with the mAb. The cDNA contained 1629 bases and encoded a 59,231-Da protein (Lys60) of 530 amino acids containing two sites for asparagine-linked core oligosaccharides. These two cDNAs showed identities of 60.3% in nucleotide sequences and 23.6% in amino acid sequences. Lys45 and Lys60 appeared to share XXEFQK as a common antigenic domain. The amino acid sequence of the Lys45 protein showed 17.4% identity and 40.9% similarity to that of PICP from Pseudomonas sp. 101. On the other hand, Lys60 showed a 24.3% identity and 51.9% similarity with human Iysosomal PICP in the amino acid sequence. A putative active center for serine protease, GTS＊xxxxxFxG, was found to be conserved among PICP homologues. The two PICPs are the first reported enzymatic markers for late Iysosomes.

• IMMUNE NETWORK
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• 제4권2호
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• pp.81-87
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• 2004

Background: In the thymus, developing thymocytes continually interact with thymic epithelial cell components. Self MHC restriction of mature T cells are imposed in the thymus through interaction of immature double positive thymocytes and thymic cortical epithelial cells. The site of negative selection, however, is a matter of debate. Through systemic injection of anti-TCR antibody or antigenic peptides, investigators suggested that most of the negative selection occurs in the thymic cortex. But the requirements for negative selection, i.e cellular counterparts and costimulatory molecules are more available in the medulla or cortico-medullary junction rather than in the thymic cortex. Methods: The direct and indirect pathways of thymocyte death after systemic anti-TCR antibody injection were separated through several experimental systems. B6 mice were either adrenalectomized or sham-adrenalectomized to evaluate the role of endogenous glucocorticoids from adrenal gland. Role of TNF were evaluated through using TNF receptor double knockout mice. Results: We found that without indirectly acting mediators such as $TNF-\alpha$ or corticosteroid, double positive thymocyte death were minimal by systemic injection of anti-TCR antibody in TNF receptor double knockout neonatal mice. Also by analyzing neonatal wild-type mice with adoptively transferred mature T cells, only peripheral activation of mature T cells could induce extensive double positive thymocyte death. Conclusion: Thus, systemically injected anti-TCR antibody mediated thymocyte death are mostly induced through indirect pathway.

• 대한바이러스학회지
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• 제27권2호
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• pp.209-216
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• 1997

Three dimensional structures of envelope protein from Korean isolates and Nakayama-NIH strain of Japanese encephalitis virus (JEV) were deduced by a computer program (HyperChem 4.0 Chemplus 1.0) based on the data of the three dimentional structure of Tick-borne encephalitis virus. In the three dimensional structure of envelope protein, neutralizing epitope and T-helper cell recognition site of C-terminal region of Korean isolates were structually similar to those of Nakayama-NIH but the N-terminal region was not. Korean JE isolates were compared with Nakayama-NIH strain by using cross-neutralization antibody test. Neutralizing activities of Korean isolates derived from guinea pigs were higher than those of Nakayama-NIH strain against Korean isolates, although the polyclonal antibody titers of Nakayama-NlH showed 1:160 to 1:640 against Korean isolates. According to the results from three dimentional structures and cross-neutralization analyses, the antigenic difference between Korean JE isolates and Nakayama-NIH strain may be dependent on structural difference of envelope protein.

• 병원약사회지
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• 제35권4호
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• pp.400-408
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• 2018

Background : Palivizumab is an intravenous monoclonal antibody which is used in the prevention of respiratory syncytial virus (RSV) infection. It is currently recommended for infants who are at high-risk for RSV infections due to preterm birth or other medical conditions such as congenital heart disease. Palivizumab is a humanized monoclonal antibody directed against an epitope in the antigenic site A of the protein F of RSV particles. Palivizumab is given once a month via intramuscular (IM) injection throughout the duration of the RSV season. Since palivizumab is known to have preventive effects against RSV infection for children with older siblings, the insurance coverage for palivizumab was expanded in October 2016. Methods : The electronic medical records of children under 2 years old who have older siblings who visited or were admitted to the Severance Hospital from October 2015 to May 2016 and from October 2016 to May 2017 were reviewed retrospectively. The data were then divided into two groups depending on the pilivizumab administration. Results : A total of 67 patients were enrolled in this study. The effectiveness in the reduction of hospitalization was statistically significant (p=0.009). Palivizumab decreased respiratory symptoms such as cough, rhinorrhea, and fever in patients with older siblings (p 0.05). Conclusions : In this study, palivizumab administration was effective in preventing RSV infection in infants with older siblings. Expanding palivizumab-prophylaxis administration to infants with older siblings may be effective in the prevention of upper respiratory infections.

• 한국동물위생학회지
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• 제25권3호
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• pp.245-257
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• 2002

The gene encoding the HN protein from the CBP-1 strain, a heat stable Newcastle disease virus (NDV) isolated from diseased pheasants in Korea, was characterized by reverse transcriptase- polymerase chain reaction(RT-PCR) and the nucleotide and amino acid sequences were analyzed following cloning of the HN gene. In all of the NDV strains studied, a 1.75 kb size cDNA fragment for the HN gene was generated by RT-PCR and smaller specific band sizes harboring the internal portions of the HN gene were also detected by using four pairs of primers. The RT-PCR was sensitive enough to detect viral transcripts when the virus titer was above 25 hemagglutination units. The amplified 1.75 kb cDNA was cloned into a BamHI site of the pVL1393 Baculo transfer vector. The nucleotide sequences of the 1,758 bp HN gene from the CBP-1 strain were determined by the dye terminator cyclic sequencing method. The gene sequences were compared among the strains of CBP-1, Texas GB, Beaudette C, LaSota, B1 and Ulster. The homology of the CBP-1 HN gene to other HN variants was 97.8% to Texas GB, 98.4% to Beaudette C, 95.4% to LaSota, 95.6% to B1 and 90.2% to Ulster. As the deduced 577 amino acid sequences were compared among the strains, the homology for CBP-1 HN appeared to be 96.7% to Texas GB, 97.9% to Beaudette C, 95.5% to LaSota, 95.5% to B1 and 92.7% to Ulster. It was evident that the amino acid sequences included 5 sites for N-asparagine linked glycosylation and 12 cysteine residues. The three conserved leucine residues within the predicted transmembrane domain of the HN protein are amino acid 30, 37 and 44. The three antigenic sites on the HN protein of NDV are amino acids 347(Glu), 481(Asn) and 495(Glu). These data indicate that the genotype of the CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than it is for the LaSota, B1 and Ulster strains.

• 미생물학회지
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• 제47권4호
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• pp.342-347
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• 2011

인체의 바이러스 감염 방어기전에서 T 림프구는 중요한 역할을 한다. 하지만, 만성 C형 간염 바이러스의 일차적 복제장소인 간염 환자의 간에서 분리된 HCV 특이 T 림프구는 심각한 기능결핍을 보인다. 이러한 T 림프구 기능결핍의 이유로는 PD-1, CTLA-4 등 면역억제 물질의 발현, 또는 간에서 특이적으로 유도되는 면역내성(immune tolerance)이 있으나, 간세포(hepatocytes)와 HCV 특이 T 림프구의 상호작용에 대해서는 명확하게 확립되어 있지 않다. 따라서 본 연구에서는 HLA(human leukocyte antigen) A2+ 간암세포주(human hepatoma cell line; huh7.5)가 항원제시(antigen presentation)를 통해 효과적으로 HCV 특이 T 림프구를 활성화시키며 간암세포주(huh7.5) 표면의 PD-L (program death ligand) 1 발현은 T림프구의 활성을 감소시켜 면역조절의 가능성이 있음을 시사하였다. 또한, HCV 특이 tetramer 반응은 T 림프구의 과도한 활성으로 자기사멸(apoptosis)의 경로에 있음을 caspase 3 활성으로 확인하였고, 역시 PD-L1의 발현이 T 림프구를 자기사멸(apoptosis)로부터 구제하여 caspase 3 활성이 감소하는 것을 확인하였다. 이는 PD-L1과 간성(liver) T 림프구 표면의 PD-1 결합이 T 림프구의 자기사멸을 막고, 또한 그 기능을 회복시켜 만성 C형 간염 치료에 응용될 수 있음을 시사한다.

• 한국가축번식학회지
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• 제9권2호
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• pp.133-139
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• 1985

An immunoassay may be defined as an analytical procedure involving the competitive reaction between a limiting concentration of specific antibody and two populations of antigen, one of which is labelled or immobillized. The advent of immunoassay has revolutionised our knowledge of reproductive physiology and the practice of veterinary and clinical medicine. Radioimmunoassay (RIA) was the first of these methods to be developed, which meausred the analyte with good sensitivity, accuracy and precision (1,2). The essential components of RIA are:-(i) a limited concentration of antibodies, (ii) a reference preparation, and (iii) an antigen labelled with a radioisotope (usually tritium or iodine-125). Most procedures invelove isolating the antibody-bound fraction and measuring the amount of labelled antigen. Good facilities are available for scintilltion counting, data reduction nd statistical analysis. RIA is undergoing refinement through:-(i) the introduction of new techniques to separate the antibody-bound and free fractions which minimize the misclassification of labelled antigen into these compartments, and the amount of non-specfic binding. (3), (ii) the development of non-extration for the measurement of haptens (4), (iii) the determination of a, pp.rent free (i.e. non-protein bound) analytes (5), and (iv) the use of monoclonal antibodies(6). In 1968, Miles and Hales introduced in important new type of immunoassay which they termed immunora-diometric assay (IRMA) based on t도 use of isotopically labelled specific antibodies(7) in a move from limited to excess reagent systems. The concept of two-site IRMAs (with a capture antibody on a solid-phase, and a second labelled antibody to a different antigenic determinant of the analyte) has enabled the development of more sensitive and less-time consuming methods for the measurement of protein hormones ovar wide concentration of analyte (8). The increasing use of isotopic methos for diverse a, pp.ications has exposed several problems. For example, the radioactive half-life and radiolysis of the labelled reagent limits assay sensitivity and imposes a time limit on the usefulness of a kit. In addition, the potential health hazards associated with the use and disposal of radioactive cmpounds and the solvents and photofluors necessary for liquid scientillation counting are incompatable with the development of extra-laboratory tests. To date, the most practical alternative labels to radioisotopes, for the measurement of analytes in a concentration > 1 ng/ml, are erythrocytes, polystyrene particiles, gold sols, dyes and enzymes or cofactors with a visual or colorimetric end-point(9). Increased sensitivity to<1 pg/ml may be obtained with fluorescent and chemiluminescent labels, or enzymes with a fluorometric, chemiluminometric or bioluminometric end-point. The sensitivity of any immunoassay or immunometric assay depends on the affinity of the antibody-antigen reaction, the specific activity of the label, the precision with which the reagents are manipulated and the nonspecific background signal (10). The sensitivity of a limited reagent system for the measurement of haptens or proteins is mainly dependent upon the affinity of the antibodies and the smalleest amount of reagent that may be manipulated. Consequently, it is difficult in practice to improve on the sensitivity obtained with iodine-125 as the label. Conversely, with excess reagent systems for the measurement of proteins it is theoretically possible to increase assay sensitivity at least 1000 fold with alternative luminescent labels. To date, a 10-fold improvement has been achieved, and attempts are being made to reduce the influence of other variables on the specific signal from the immunoreaction.