• The Microorganisms and Industry
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• v.17 no.1
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• pp.38-50
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• 1991

Flow cytometer(유식세포분석분리기:FCM)는 부유되어 있는 세포의 여러 특성을 측정할 수 있도록 고안되었기 때문에 동물의 체내에 있는 세포중 자연적으로 부유되어 있는 혈구세포를 분석하기가 가장 용이하다. 면역계에서 가장 중추적인 역할을 담당하며 면역반응에 특이성과 기억능력 등을 부여하는 림프구와 탐색작용을 가장 왕성히 나타내는 과립구나 단구 등이 혈액내에 존재하기 때문에 FCM을 이용하여 개체의 방어기능을 맡고 있는 면역세포를 분석하는 일이 가장 먼저 활발히 이루어졌음은 당연하였다. 도한 1970년대 면역학이 큰 발전에 이룩한 단크론항체가 Miller와 Kohler에 의하여 개발되었기 때문에 면역세포의 분화항원에 대한 여러 종류의 단크론항체는 FCM의 이용으로 면역세포를 여러 아형으로 분석하고 분리가 가능하게 하였다. 마지막으로 FCM이 최근에 개발된 것에 맞추어 세포성 면역학이 30년 전부터 급속도로 연구되었으므로 면역학적 연구가 FCM의 이용으로 더욱 활성화되었음은 주지의 사실이다. 이런 시점에서 저자는 FCM의 원리와 면역생물학에서의 FCM의 역할에 대하여 간단히 소개하려고 한다.

• The Korean Journal of Malacology
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• v.25 no.2
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• pp.135-143
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• 2009

The Manila clam, Ruditapes philippinarum, has been considered as a sentinel species due to dominant distribution along the coast of Korea and well developed regulatory system. In order to develop and understand immune responses of the Manila clams, clams were exposed to $50\;{\mu}g/L$ of cadmium chloride (Cd) for 8 days and monitored the cellular immune parameters of the hemocytes including blast cell composition, DNA damage, necrosis, apoptosis and hemocyte mortality using a flow cytometer. The results showed that all immune parameters analyzed in the present study increased remarkably compared to the controls and the increases were statistically significant. Apoptosis rate was higher than necrosis rate in the clams exposed to Cd suggesting that apoptosis was preferably induced by the concentration of Cd used in the present study. Our study indicates that the measurement of cellular immune responses of the Manila clam using flow cytometer will be a useful technique for assessment of heavy metal contamination in marine environment.

• Applied Microscopy
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• v.32 no.4
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• pp.385-398
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• 2002

The purpose of the present study is to investigate whether stromal cells supporting specific microenvironment for hematopoiesis of bone marrow are affected by toxicants and therapeutic drugs such as antibiotics and anticancer drugs and whether laminin-1 is associated with such effects. SD rats were intraperitoneally injected with 75 mg/kg of cyclophosphamide which is widely used to treat infant's solid tumor, leukemia and myeloma and sacrificed after 3 days, 1 week, 3 weeks or 5 weeks of injection. The bone marrow extracted and paraffin-sectioned was analyzed using immunohistochemical staining. A part of tissues was subjected to electron microscopy following reaction with rabbit anti-laminin antibody, biotinylated goat anti-rabbit IgG conjugated with 12 nm gold particles, and staining with uranyl acetate. 1. The bone marrow tissue at day 3 post injection with cyclophosphamide displayed dilated venous sinus, partial necrotic death, and decreased number of hematopoietic cells. Laminin-1 was intensively stained in the reticular and adipose tissues. 2. Up to 5 weeks post injection, laminin-1 was stained at a low level in the stromal tissue of bone marrow and the number of hematopoietic cell was increased. 3. Deposition of the gold particle which represents laminin-1 expression was observed at the highest level in the stromal cells of bone marrow obtained 3 days after injection, and decreased after 1 to 5 weeks. These results suggest that stromal cells which play a role in supporting microenvironment for bone marrow hematopoiesis augment induction of laminin-1 expression and activation upon administration of cyclophosphamide.

• The Korean Journal of Zoology
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• v.35 no.4
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• pp.405-413
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• 1992

등검은메뚜기(EuprepoceRneTmis shirak시 성충에서 면역작용에 관여하는 혈구로 확인된 Plasmatocvte와 granu10cvte 1 딪 granu10cyte 11의 형태와 미세구조적 특징을 광학현미경과 전자현미경을 이용하여 관찰하였다. Plasmatocyte는 방추형 또는 맡은 세포질돌기를 형성하고 있는 다형의 형태로 세포질에는 리소조옴과 식포가 존재하고 있었고, 식포의 융합에 의해서 형성된 multivesicular body가 관찰되었다. Granulocyte 1은 구형의 형태로 세포질에는 약 0.2-0.3 urn 크기의 과립을 함유하고 있었으며, 특히 원형질막 가장자리에 세포의 골격을 유지하는 미세소관 다발을 갖고 있었다. Granulocyte 11는 난형 또는 방추형의 형태로 세포질에는 약 0.5-0.8 Um의 과립이 세포질의 대부분을 차지하고 있었으며, 세포소기 관은 granu10cyte 1보다 덜 발달되어 있었다.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.162-162
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• 1993

제 3세대 platinum complex인 SKI 2053R에 의한 Salmonella typhimurium의 복귀변이빈도, CHL세포(Chinese Hamster Lung)에 대한 염색체이상 유발율 및 ddY계 마우스에서의 골수분화세포에 대한 염색체이상유발로 기인한 소핵의 빈도수를 관찰하여 SKI 2053R의 유전독성을 평가하였다. Salmonella typhimurium를 이용한 복귀돌연변이시험에서 SKI 2053R은 매우 경미한 정도의 돌연변이 유발성을 가지는 것으로 판단되며, 균주 특이적 돌연변이 유발성으로 보아 염기쌍치환형의 돌연변이를 유발하는 것으로 사료되며 포유류배양세포를 이용한 염색체이상실험에서 대사 활성 부재 및 존재하의 모든 시험 농도에서 10％ 이상의 염색체이상을 가진 세포가 관찰되었으며 염색체 상의 종류로는 염색분체형 교환 (cse)이 가장 많이 관찰되었다. 설치류를 이용한 소헥시험에서는 ddY계 마우스 골수세포의 분화과정에서 염색체이상을 유발하며, 다염성적혈구의 정염성적혈구에 대한 출현비율이 감소하는 결과로 볼때 방추체기능의 저해를 일으키는 것으로 판단된다. 그러므로 본 시험조건데 있어서 SKI 2053R은 소핵을 유발하는 물질로 결론지었다.

• Journal of fish pathology
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• v.13 no.2
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• pp.129-136
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• 2000

To understand host-defence mechanism of clam(Ruditapes philippinarum) hemocyte against foreign materials, classification and their seasonal change in the number were performed. clams collected from a farm in Julpo Bay, Gochang, Chollabuk-Do were used in this experiment. Lots of hemocytes were found between the muscle fibers and connetive tissue of posterior adductor muscle. Hemocytes of R. philippinarum were classified into granulocytes and agranulocytes. Granulocytes were composed of three types, basophilic granulocyte, acidophilic granulocyte and fibrocyte in accordance with the staining affinities of their cytoplasmic granules. Fibrocyte has filopodia and vesicle in endoplasm and bigger than other granulocytes in size. Agranulocytes were less in the number and smaller in the size compared to those of granulocytes. Hyalinocytes had no granule in their cytoplasm. The nucleus located in the center of the cell was oval or spherical shaped. In electron microscopic observation, granulocytes and hyalinocytes contained electron-dense vesicles and some small lucid vesicles in their cytoplasm, respectively. Granulocytes phagocytosed more zymosan particles than hyalinocytes. Acidophilic granulocytes showed higher phagocytic ratio than basophilic granulocytes. Total hemocyte numbers showed the highest at April to August and the lowest at October to December. In the composition of each hemocyte, basophilic granulocytes were always more than acidophilic granulocytes and hyalinocytes.

• Korean journal of applied entomology
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• v.51 no.4
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• pp.349-356
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• 2012

Eicosanoid mediates various cellular immune responses in insects. This study aimed to discover its novel action on the modulation of hemocyte populations in response to an immune challenge. Upon bacterial challenge, the last instar larvae of the beet armyworm, Spodoptera exigua, increased their total hemocyte density in 2 h, and then decreased it to a basal hemocyte density level. This rapid increase in total hemocyte density was explained by an increase of plasmatocyte and spherulocyte densities. When larvae were treated with dexamethasone (a specific phospholipase $A_2$ ($PLA_2$) inhibitor), they did not show any increase in hemocyte density in response to bacterial challenge. However, the addition of arachidonic acid (a catalytic product of $PLA_2$) to larvae treated with dexamethasone recovered the up-regulation of hemocyte density in response to bacterial infection. Among eicosanoid, cyclooxygenase (COX), but not lipoxygenase (LOX), products seemed to mediate the increase of hemocyte density in response to bacterial infection because naproxene (a COX inhibitor) inhibited the hemocyte density increase, though esculetin (a LOX inhibitor) did not. Prostaglandin $E_2$, a COX product, significantly increased the hemocyte density even without bacterial infection. These results suggest that eicosaniod mediates a rapid increase in total hemocyte density in response to immune challenge.

• Clinical and Experimental Pediatrics
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• v.45 no.2
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• pp.247-255
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• 2002

Purpose : This study was undertaken to obtain basic data about the megakaryocyte colony formation of fetal liver cells by using immunocytochemical staining and ex vivo culture with growth factors. Methods : The mononuclear cells were isolated from fetal liver and bone marrow with idiopathic thrombocytopenic purpura(ITP) and pancytopenia. These mononuclear cells were cultured in $MegaCult^{TM}-C$(Stem Cell Tech, Canada) media in the presence of growth factors and CFU-Megakaryocyte( CFU-Mk) colonies were counted on day 12. The expansion of CD34+ and CD41+ cell was analyzed by flow cytometry after 5 days incubation using flask culture. Results : The numbers of CFU-Mk colonies of mononuclear cells obtained from fetal liver in the 11th week gestational age were more than those in the 19th week specimens; growth factors could not enhance the colony expansion in all cases. Total numbers of CFU-Mk colony of fetal liver cells were higher than bone marrow from ITP or pancytopenia groups. The numbers of pure or large CFU-Mk colonies of fetal liver cells were also higher than bone marrow specimens. The rate of CD34+ cell expression of fetal liver was increased after flask culture and the enhancement effect of epression was seen only in cases which added thrombopoietin. The rate of CD41+ cell expression of fetal liver was increased after incubation, but the enhancement effect of growth factors was unclear. Conclusion : This study revealed good results about the megakaryocyte colony assay of fetal liver mononuclear cells using $MegaCult^{TM}-C$ media. This study suggests that the fetal liver could be a good source of megakaryocytic progenitor cells for clinical application in hematopoietic stem cell transplantation.

• The Korean Journal of Pesticide Science
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• v.13 no.2
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• pp.111-116
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• 2009

Juvenile hormone (JH) is an insect hormone mediating immature metamorphosis and adult reproduction. It also mediates immune responses to suppress hemocyte behavior, which is, however, activated by ecdysteroid. This study investigated an effect of a commercial pyriproxyfen (a JH agonist) formulation on a cellular immune response of the diamondback moth, Plutella xylostella, and analysed its mixture with Bacillus thuringiensis (Bt) in insecticidal potency. The commercial pyriproxyfen formulation significantly suppressed hemocyte-spreading behavior at low doses as did in pyriproxyfen technical grade. When the commercial pyriproxyfen formulation was mixed with Bt, Bt toxicity was significantly increased against P. xylostella larvae in laboratory. The mixture effect was then confirmed in field cultivating cabbage infested with P. xylostella larvae. The mixture showed a significantly enhanced mortality and reduced effective lethal time, compared to only Bt treatment.

• Development and Reproduction
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• v.5 no.2
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• pp.93-100
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• 2001

Precise mechanism by which the fetus can escape from mother's immune rejection is not well understood yet over the last 50 years. The clarification of immune mechanism at the feto-maternal interface is very important, because this can be a common pathogenesis of various pathologic conditions including spontaneous abortion, habitual abortion fetal growth restriction preeclampsia, implantation failure after assisted reproductive techniques, and fetal death. In this review, current hypothetical contents were described with the priority of importance: 1) The center of this mechanism is cross-talk between the expression of HLA-C, E, G on the extravillous cytotrophoblasts and their receptors on decidual NK cell, 2) immunomodulation, 3) innate immunity is the main immunologic mechanism, 4) various mechanisms besides HLA system(eq. complement) may be associated. The overall balance of immunomodulation among these mechanisms should result in the outcome of each pregnancy. Further researches regarding the regulation of HLA system, roles of cytokines, complements should be followed in the future.