• 제목/요약/키워드: antibody formation

검색결과 188건 처리시간 0.025초

Pseudomonas tolaasii 박테리오파지에 특이적인 다클론항체 형성 및 이를 이용한 파지 교차 반응성 (Pseudomonas tolaasii bacteriophage-specific polyclonal antibody formation and its cross reactivity to various phages)

  • 윤영배;박수진;김영기
    • Journal of Applied Biological Chemistry
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    • 제62권3호
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    • pp.287-292
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    • 2019
  • Pseudomonas tolaasii는 느타리버섯에 갈반병을 일으키는 병원균주로, 다양한 변이균주들을 분리하여 $P1{\alpha}$$P1{\beta}$, $P1{\gamma}$ 세 가지 소그룹으로 분류하였다. 각 그룹별 균주들에 특이적인 박테리오파지를 이용한 파지테라피는 갈반병 방제에 매우 성공적이었다. 본 연구에서는, 박테리오파지들의 특성을 구명하기 위하여 소그룹별 대표균주을 이용하여 파지를 분리하였고, 이들의 다클론항체를 제작하여 파지들 사이에 유연관계를 조사하였다. 파지 준비물은 $10^{10}pfu/mL$ 이상으로 토끼의 다리 근육에 주사하였고, 3회의 반복주사에 의해 다클론항체가 얻어졌다. 파지 ${\phi}6264$에 대한 항체의 역가는 $2{\times}10^7Ab/mL$ 이상, 파지 ${\phi}HK2$에 대해서는 $1{\times}10^6Ab/mL$, 파지 ${\phi}HK19$${\phi}HK23$에 대해서는 $1{\times}10^7Ab/mL$ 이상이었다. 항체와 이에 특이적인 파지 사이에는 매우 높은 반응특이성이 있었고, 소그룹이 다른 파지의 항체와 파지 사이에도 일부 교차반응성을 확인하였다. 파지 ${\phi}6264$에서 생성된 $Ab{\phi}6264$는 모든 $P1{\alpha}$ 소그룹의 파지들과 반응성을 보였으나, 파지 ${\phi}HK16$을 제외한 다른 소그룹의 파지들과는 반응하지 않았다. $P1{\gamma}$ 소그룹에서 생성된 $Ab{\phi}HK23$$P1{\beta}$ 소그룹의 모든 파지들을 불활성화시켜 가장 넓은 항체범위를 보였다. 항체와 파지를 이용한 숙주균과의 관계를 분석하였을 때, 16S rRNA 유전자 분석에 의한 숙주균의 근연관계와 항체를 이용한 숙주균의 파지들 사이의 구조적 근연관계는 상당히 차이가 있음을 확인하였다. 결론적으로, 박테리오파지의 숙주균 특이성과 항체를 이용해 측정한 파지의 껍질단백질 구조유사성 사이에는 약한 상관성을 보였다.

전씨백출산(錢氏白朮散)이 생쥐의 체액성(體液性) 면역반응(免疫反應)과 세포성(細胞性) 면역반응(免疫反應)에 미치는 효과(效果) (Effect of Junsibaekchulsan on the humoral and cell-mediated immune responses in mouse)

  • 심문경;박은정
    • 대한한방소아과학회지
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    • 제8권1호
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    • pp.39-58
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    • 1994
  • Even though appropriate immune response is necessary for the survival of the individual, excessive or insufficient immune response might cause autoimmune or allergic disease respectively. So the immune response must be controlled to the degree that is beneficial for the well being of the individual. This study was undertaken to know the effects of Junsibaekchulsan(JB) on the immune system od the mouse. For the evalulation of the cell-mediated immunity(CMI), delayed-type hypersensitivity against dinitrofluorobenzene(DNFB) were measured, and humoral immunity, hemagglutinin and hemolysin titers against SRBCs(sheep red blood cells) were measured, and rosette formation of spleen cells with SRBCs were measured. For the evaluation of innate immunity, phagocytic activity of macrophages, natural killer cell activity, and reactive nitrogen and oxygen intermediates were measured. The results are as follows: 1. The administration of JB depressed the antibody formation (hemagglutinin and hemolysin) against SRBCs. 2. The administration of JB did not affect the delayed-type hypersensitivity against DNFB. 3. The administration of JB did not affect the cytotoxic activity of natural killer cells. 4. The administration of JB increased the phagocytic activity of macrophages. 5. The administration of JB increased the rosette formating cells of the spleen cells. 6. The exposure of JB induced the secretion of reactive nitrogen intermediates but administration of JB deperssed the production of reactive oxygen intermediates. Administration of JB selectively depressed the humoral immune response without affecting CMI and innate immunity. These results of JB on the immune system might be useful for the treatment of such.

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누에의 變態에 따른 貯藏蛋白質의 出現과 分布에 관하여 (On the Occurrence and Distribution of Storage Proteins During the Metamorphosis of Bombyx mori L)

  • Eul Won Seo;Hak Ryul Kim
    • 한국동물학회지
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    • 제29권1호
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    • pp.1-12
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    • 1986
  • 누에의 變態期 동안 貯藏蛋白質의 出現과 번데기 시기동안 각 組織에 따른 貯藏蛋白質의 分布를 살펴보기 위해 電氣泳動法, 免疫學的 方法 및 column chromatography法을 使用하였다. 혈림프의 貯藏蛋白質은 2개로 區分이 되었고, 5령 初期부터 出現하고 있으며 지방체 단백질과도 동일한 전기영동상의 移動度를 갖고 있다. 이의 量的 變化는 終令期에는 혈림프에서 높은 濃度를 유지하다.  化後에는 脂肪體에 축적이 되는 경향을 나타내며 특히 저장단백질-2가 암수에서 모두 두드러진 저장단백질의 양상을 보이고 있다. 이러한 貯藏蛋白質은 종령 末期에는 큐리클 단백질 형성에도 관여하는 것 같으며 번데기에는 中腸도 일시적으로 저장단백질을 저장하는 것 같다. 또한 貯藏蛋白質中 저장단백질-2는 vitellogenin과 정기영동 및 면역학적으로 동일한 이동도를 나타내고 있으며 특히 번데기시기동안 卵黃蛋白質의 항체에 대해 항원-항체반응을 나타내고 있어 卵形成過程에도 밀접하게 관여하는 것 같다.

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Angiotensin II Promotes Smooth Muscle Cell Proliferation and Migration through Release of Heparin-binding Epidermal Growth Factor and Activation of EGF-Receptor Pathway

  • Yang, Xiaoping;Zhu, Mei J.;Sreejayan, N.;Ren, J.;Du, Min
    • Molecules and Cells
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    • 제20권2호
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    • pp.263-270
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    • 2005
  • Transactivation of EGF-receptor (EGFR) by G-protein coupled receptors (GPCRs) is emerging as an important pathway in cell proliferation, which plays a crucial role in the development of atherosclerotic lesion. Angiotensin II (Ang II) has been identified to have a major role in the formation of atherosclerotic lesions, although the underlying mechanisms remain largely unclear. We hypothesize that Ang II promotes the proliferation and migration of smooth muscle cells through the release of heparin-binding epidermal growth factor like growth factor (HB-EGF), transactivation of EGFR and activation of Akt and Erk 1/2, with matrix metalloproteases (MMPs) playing a dispensable role. Primary rat aortic smooth muscle cells were used in this study. Smooth muscle cells rendered quiescent by serum deprivation for 12 h were treated with Ang II (100 nM) in the presence of either GM6001 ($20{\mu}M$), a specific inhibitor of MMPs or AG1478 ($10{\mu}M$), an inhibitor of EGFR. The levels of phosphorylation of EGFR, Akt and Erk 1/2 were assessed in the cell lysates. Inhibition of MMPs by GM6001 significantly attenuated Ang II-stimulated phosphorylation of EGFR, suggesting that MMPs may be involved in the transactivation of EGFR by Ang II receptor. Furthermore Ang II-stimulated proliferation and migration of smooth muscle cells were significantly blunted by inhibiting MMPs and EGFR and applying HB-EGF neutralization antibody, indicating that MMPs, HB-EGF and EGFR activation is necessary for Ang-II stimulated migration and proliferation of smooth muscle cells. Our results suggest that inhibition of MMPs may represent one of the strategies to counter the mitogenic and motogenic effects of Ang II on smooth muscle cells and thereby prevent the formation and development of atherosclerotic lesions.

방사선조사와 저칼슘식이가 백서구치의 상아질과 백악질형성에 미치는 영향에 관한 실험적 연구 (THE EFFECTS OF IRRADIATION AND CALCIUM-DEFICIENT DIET ON DENTIN AND CEMENTUM FORMATION OF RAT MOLAR)

  • 신재창;황의환;이상래
    • 치과방사선
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    • 제28권1호
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    • pp.87-109
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    • 1998
  • The present study was designed to elucidate the effects of the Co-60 γ irradiation and/or calcium­deficient diet on the dentin and cementum formation of rat molar. The pregnant three-week old Sprague­Dawley rats were used for the study. The experimental group was divided into two groups, irradiation/normal diet group and irradiation/calcium-deficient diet group. The control group was non­irradiation/normal diet group. The abdomen of the rats at the 19th day of pregnancy were irradiated with single absorbed dose of 350cGy. The rat pups were sacrificed on the 14th day after delivery and the maxillae including molar tooth germ were taken. The specimens including the 1st molar tooth germ were prepared to make tissue sections for light and transmission electron microscopy. Some of tissue sections for light microscopy were stained immunohistochemically with anti-fibronectin antibody. The results were as follows; 1. The Hertwig's epithelial root sheath cells, which are related to the differentiation of the tooth-forming cells, showed irregular cellular arrangement, decrease of intercellular junctional complex, and decreased immunoreactivity to the fibronectin after irradiation. These were more severe in the irradiation/calcium-deficient diet group. 2. The cementoblasts at the cementum-forming area showed chromatin clumpings after irradiation. The immu noreactivity to the fibronectin was weaken after irradiation, especially irradiation/calcium-deficient diet group. 3. The odontoblasts at the dentin-forming area showed increase of lysosomes in the cytoplasm and destruction of intercellular junctional complex. The irradiation/calcium-deficient diet group showed decrease of number and density of the electron dense particles and a large number of vacuoles scattered in the dentin matrix. The immunoreactivity was weaken.

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성견 하악골 절단 후 기계적 골 견인에 의해 형성된 골 신장부에 대한 시기별 조직학적 변화 (HISTOLOGICAL CHANGES IN THE ELONGATED BONE AFFECTED BY OSTEODISTRACTION OF THE MANDIBLE IN THE DOG)

  • 백선호;안병근;박영주;박희건;박준우;이건주;이용찬;조병욱
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제27권5호
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    • pp.404-416
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    • 2001
  • Purpose : Traditionally, the treatement of choice has been a bone grafting procedure to increase the length of bone in case of actual length discrepancy. But, bone grafting procedure has many disadvantages, for example, graft resorption, donor site morbidity, and so on. So, many trials have been performed to avert the use of autogenous bone graft via introducing new materials or methods. And, one of those trials has been realized by the development of a technique inducing bone lengthening by osteotomy (or corticotomy) and slow gradual distraction of the osteotomized segments. This new technique of bone lengthening dates back to the early 20th century. But, the majority of information concerning the biology of new bone formation during bone lengthening and technical details of the procedure were produced by extensive clinical and experimental studies performed by Ilizarov, a Russian surgeon. According to Ilizarov, with adequate blood supply, preservation of periosteum, rigid fixation of the osteotomized segments, and proper rate and rhythm of distraction, intramembranous bone rapidly develops within the distraction gap in the limb lengthening procedure. In the limb lengthening, many orthopedic surgeons try to observe the biologic and clinical principles recommended by Ilizarov. In the oral and maxillofacial region, however, not a few studies must be performed to apply this surgical technique in the clinical cases. Besides, the mechanism of bone formation in the distraction gap is not clear, yet. The purpose of this experiment was to scrutinize serially the histological changes in the elongated bone affected by osteodistraction of the mandibular body in an adult canine model. In addition, it was performed to confirm the presence of specific region(s) which was important in the bone formation in the gap through the observation of the expression pattern of osteocalcin and osteonectin with the immunohistochemical examination. Materials and Methods : The experimental and control specimens were obtained from seven adult male mongrel dogs weighing over 20kg. The distractors were custom-made linear extraoral devices and bicortical fixation screws were 2.3mm in diameter, 50mm in total length, 15mm in screw length. The distractors were devised to produce a linear gap of 0.75mm between two bony segments every $360^{\circ}$ turn of the rotation rod of the device. The mandibular body of the right side of each animal was corticotomized perpendicular to the occlusal plane and then two bony segments were separated completely by careful manipulation of the segments with bone forceps. The left side of each animal was left intact. This side was served as control. At sixth day after osteotomy and fixation of the segments were performed, distraction of the segments was commenced with a rate of 1.1mm/day and a rhythm of two/day for ensuing 7 days. The animals were euthanized at the 16th. 29th, and 44th day after the osteotomy. The bony specimens were decalcified, embedded in paraffin, sectioned $5{\mu}m$ thick and stained with H&E. The prepared specimens were examined under the light microscope. And, immunohistochemical examinations using anti-osteocalcin antibody (OC1, Biodesign, USA) and anti-osteonectin antibody (Haematologic Technologies Inc., Essex, VT) to locate the expressions of osteocalcin and osteonectin, respectively, were performed. Results : 1. New bone was observed already at the 16th. day after osteotomy. This suggests that new bone formation in osteodistraction was commenced at an early stage of the regenerative process. But, radiologically and microscopically, bony union was not completed in the distraction gap at the 44th. day after osteotomy. Therefore, rigid fixation must be maintained between the bony fragments till the complete bony union is confirmed clinically rather than one month or so after the completion of distraction.

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흡연특이성 발암물질이 특정 Protein Kinase C Isoform에 미치는 영향 (Effects of Tobacco-Specific Carcinogen on Protein Kinase C Isoforms)

  • 강형석;고무성;박기성;이섭;전상훈;권오춘
    • Journal of Chest Surgery
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    • 제36권9호
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    • pp.666-673
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    • 2003
  • 폐암의 주된 원인으로 알려진 흡연은 그 악성세포 발현기전이 아직 정확히 규명된 바 없다. 이에 저자들은 흡연에 의한 발암성의 지표로 흡연 중에 특이적으로 존재하는 강력한 발암물질인 NNK(4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone)를 이용하여 흡연에 따른 폐암의 발생과 그 Protein kinase C (PKC) isoform과 관련된 기전에 관한 연구를 시도하였다. 대상 및 방법: 인체 상피세포를 NNK에 노출시킨 후 saturation density, soft agar colony formation, cell aggregation 및 foci의 출현 등의 양상을 파악하여 세포 발암성 여부를 관찰하였으며 NNK를 15분간 노출시킨 후 PKC의 변화는 세포 내 PKC isoform의 양을 cytosolic fraction과 membrane fraction으로 분리하여 측정하여 분석하였다. 결과: NNK 투여군에서 saturation density, soft agar colony formation, cell aggregation 및 foci의 출현 시기 등의 세포 발암성을 뚜렷이 나타내었으며 PKC isoform분석의 경우 PKC-$\alpha$의 membrane fraction의 뚜렷한 증가를 보였으며 이러한 활성은 용량-의존적인 형태를 유지하였다. PKC-$\varepsilon$은 NNK 처리 시 용량-의존적으로 cytosol fraction의 감소 및 membrane fraction의 증가를 뚜렷하게 보였고 NNK에 의한 PKC-λ의 변화는 감지되지 않았다. 결론: 본 연구는 화학적 발암물질인 NNK가 인체발암화에 관여함을 재차 확인하면서 초기 과정에 관여하는 PKC isoform의 변화를 분석함으로써 total PKC활성이 아닌 isoform 각각에 대한 변화를 확인하였다는 점에서 앞으로 인체상피세포 기원의 폐암 생성 기전 연구에 기여할 것으로 생각한다.

멍게의 수압수용체세포 형성에서 FGF와 MEK 신호의 역할 (Role of FGF and MEK Signaling in Formation of the Hydrostatic Pressure Receptor Cells during Ascidian Embryogenesis)

  • 서형주;김길중
    • 한국발생생물학회지:발생과생식
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    • 제13권4호
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    • pp.291-296
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    • 2009
  • 멍게 유생의 뇌포에는 2개의 감각색소세포인 평형기와 안점 이외에 또 다른 감각세포로 추정되는 수압수용체세포가 존재한다. 수압수용체세포 형성에 관해서는 현재까지 거의 알려진 것이 없다. 본 연구에서는 수압수용체세포 형성에서 FGF 신호전달 과정의 관련성을 조사했다. 수정란에 Hr-FGF9/16/20 antisense MO를 미세주입했을 때, 발생한 유생에서 수압수용체세포 특이적 Hpr-1 항원의 발현이 검출되지 않았다. 32세포기부터 FGF 수용체 억제제 SU5402 및 MEK 억제제 U0126을 처리한 배아도 수압수용체세포를 갖지 못한 유생으로 발생했다. 다음으로 수압수용체세포 형성에 FGF 신호전달 과정이 관련되는 시기를 자세히 조사했다. 수압수용체세포 형성에는 FGF 수용체 활성이 16세포기부터 64세포기까지 필요하다는 것이 시사되었다. U0126은 8세포기부터 후기 낭배기까지 Hpr-1 항원 발현을 억제했다. Hpr-1 항원 발현은 신경판기 직전부터 U0126의 영향을 받지 않았다. 따라서, 멍게에서 수압수용체세포 형성은 1차 신경유도기부터 후기 낭배기까지 FGF 신호전달 과정을 필요로 한다는 것이 밝혀졌다.

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A Novel Complement Fixation Pathway Initiated by SIGN-R1 Interacting with C1q in Innate Immunity

  • Kang, Young-Sun
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2008년도 International Meeting of the Microbiological Society of Korea
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    • pp.23-25
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    • 2008
  • Serum complement proteins comprise an important system that is responsible for several innate and adaptive immune defence mechanisms. There were three well described pathways known to lead to the generation of a C3 convertase, which catalyses the proteolysis of complement component C3, and leads to the formation of C3 opsonins (C3b, iC3b and C3d) that fix to bacteria. A pivotal step in the complement pathway is the assembly of a C3 convertase, which digests the C3 complement component to form microbial-binding C3 fragments recognized by leukocytes. The spleen clears microorganisms from the blood. Individuals lacking this organ are more susceptible to Streptococcus pneumoniae. Innate resistance to S. pneumoniae has previously been shown to involve complement components C3 and C4, however this resistance has only a partial requirement for mediators of these three pathways, such as immunoglobulin, factor B and mannose-binding lectin. Therefore it was likely that spleen and complement system provide resistance against blood-borne S. pneumoniae infection through unknown mechanism. To better understand the mechanisms involved, we studied Specific intracellular adhesion molecule-grabbing nonintegrin (SIGN)-R1. SIGN-R1, is a C-type lectin that is expressed at high levels by spleen marginal-zone macrophages and lymph-node macrophages. SIGN-R1 has previously been shown to be the main receptor for bacterial dextrans, as well as for the capsular pneumococcal polysaccharide (CPS) of S. pneumoniae. We examined the specific role of this receptor in the activation of complement. Using a monoclonal antibody that selectively downregulates SIGN-R1 expression in vivo, we show that in response to S. pneumoniae or CPS, SIGN-R1 mediates the immediate proteolysis of C3 and fixation of C3 opsonins to S. pneumoniae or to marginal-zone macrophages that had taken up CPS. These data indicate that SIGN-R1 is largely responsible for the rapid C3 convertase formation induced by S. pneumoniae in the spleen of mice. Also, we found that SIGN-R1 directly binds C1q and that C3 fixation by SIGN-R1 requires C1q and C4 but not factor B or immunoglobulin. Traditionally C3 convertase can be formed by the classical C1q- and immunoglobulin-dependent pathway, the alternative factor-B-dependent pathway and the soluble mannose-binding lectin pathway. Furthermore Conditional SIGN-R1 knockout mice developed deficits in C3 catabolism when given S. pneumoniae or its capsular polysaccharide intravenously. There were marked reductions in proteolysis of serum C3, deposition of C3 on organisms within SIGN-$R1^+$ spleen macrophages, and formation of C3 ligands. The transmembrane lectin SIGN-R1 therefore contributes to innate resistance by an unusual C3 activation pathway. We propose that in the SIGN-R1 mediated complement activation pathway, after binding to polysaccharide, SIGN-R1 captures C1q. SIGN-R1 can then, in association with several other complement proteins including C4, lead to the formation of a C3 convertase and fixation of C3. Therefore, this new pathway for C3 fixation by SIGN-R1, which is unusual as it is a classical C1q-dependent pathway that does not require immuno globulin, contributes to innate immune resistance to certain encapsulated microorganisms.

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Surface Plasmon Resonance Immunosensor for Detection of Legionella pneumophila

  • Oh, Byung-Keun;Lee, Woochang;Bae, Young-Min;Lee, Won-Hong;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권2호
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    • pp.112-116
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    • 2003
  • An immunosensor based on surface plasmon resonance (SPR) onto a protein G layer by Self-assembly technique was developed for detection of Legionella pneumophila. The protein G layer by self-assembly technique was fabricated on a gold (Au) surface by adsorbing the 11-mercaptoundecanoic acid (MUA) and an activation process for the chemical binding of the free amino (-NH$_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC) in series. The formation of the protein G layer by self-assembly technique on the Au Substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The Surface topographies of the fabricated thin films on an Au substrate were also analyzed by using an atomic force microscope (AFM). Consequently, an immunosensor for the detection of L. pneumophila using SPR was developed with a detection limit of up to 10$^2$CFU per mL.