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TPA로 분화된 U937 세포에서 사람 세포거대바이러스에 의한 c-jun Promoter 활성도의 변화
박정규,김대중,김진희,한태희,황응수,최명식,국윤호,최성배,차창룡,Park, Chung-Gyu,Kim, Dae-Joong,Kim, Jin-Hee,Han, Tae-Hee,Hwan, Eung-Soo,Choi, Myong-Sik,Kook, Yoon-Hoh,Choi, Sung-Bae,Cha, Chang-Yong 대한미생물학회 1999 Journal of Bacteriology and Virology Vol.28 No.1
Transient transfection assay has been done to evaluate whether the c-jun activation would be prerequisite to the induction of permissiveness against human cytomegalovirus using in vitro cell model in which U937 has been induced to express CD11b and CD14 to become potential monocyte/macrophage cells by TPA treatment. U937 cells were treated with $10\;{\mu}M$, $50\;{\mu}M$ or $100\;{\mu}M$ of TPA. The cell morphology change was observed and the expression of the CD11b and CD14 was confirmed by FACS. Differentiated cells were transfected with pJLuc reporter vector which contained the wild type murine c-jun promoter spanning the SP1, CTF, ATF/CREB and MEF-2 binding sites upstream of the firefly luciferase gene. After 48 hrs of transfection, the cells were infected with HCMV Towne strain and the luciferase activity was assessed at 1 hand 4 h pi. The transfection assay showed no activation of the c-jun promoter at 1 h pi, instead, it showed 2 times increase of the its activity at 4 h pi. There was no difference of the c-jun promoter activation between TPA treated and untreated U937 cells, implying that c-jun activation might not be prerequisite for allowing cells to be premissive to HCMV, although HCMV infection itself could activate c-jun promoter.
황응수,조명제,차창룡,최강원,이승훈,장우현,Hwang, Eung-Soo,Cho, Myung-Je,Cha, Chang-Yong,Choe, Kang-Won,Lee, Seung-Hoon,Chang, Woo-Hyun The Korea Society for Microbiology 1986 大韓微生物學會誌 Vol.21 No.3
Serum samples from 51 patients with clinically suspected typhoid fever were tested for immunoglobulin G (IgG), IgM and IgA antibodies against the whole bacteria antigen of Salmonella typhi by an enzyme-linked immunosorbent assay. The levels of IgG and IgA antibody to-whole bacteria antigen were higher in the culture-proven patients than in controls. The levels of IgM antibody to- whole bacteria antigen showed better discrimination between culture negative patients and controls than those of IgG or IgA antibody to-whole bacteria antigen. The enzyme-linked immunosorbent assay was much more sensitive than the Widal test. It would be a useful tool for the diagnosis of typhoid fever with a single serum sample. 장티푸스가 의심되는 51명의 환자 혈청내에서 Salmonella typhi의 균체항원에 대한 IgG, IgM과 IgA 항체률 효소면역측정법으로 측정하였다. 균체항원에 대한 IgG와 IgA 항체가는 세균이 분리되어 장티푸스로 확진된 환자에서 건강대조군보다 높았다. 반면 세균을 분리하지는 못하였지만 임상증상 등으로 장티프스를 의심하는 환자에서는 IgM항체가는 건강대조군보다 높았다. 균체항원에 대한 항체를 효소면역 측정법으로 측정하는 것이 Widal 검사보다 민감도가 높았다. 장티푸스를 진단하는데 균체항원을 부착시켜 효소면역측정법을 사용하는 것이 유용할 것이다.
장티푸스환자의 혈청내 Salmonella typhi에 대한 IgG subclass항체의 분포
김영중,황응수,강재승,차창룡,장우현,김윤원,조민기,민창홍,Kim, Young-Jung,Hwang, Eung-Soo,Kang, Jae-Seung,Cha, Chang-Yong,Chang, Woo-Hyun,Kim, Yoon-Won,Cho, Min-Ki,Min, Chang-Hong 대한미생물학회 1986 大韓微生物學會誌 Vol.21 No.4
To diagnose the typhoid fever rapidly and accurately in clinically suspected patients, the levels of IgG subclass antibody were measured by enzyme-linked immunosorbent assay(ELISA). With symptom, blood culture and agglutination test, tested persons were categorized into 6 groups as typhoid fever, FUO, paratyphi A or B, other bacterial infctions, cancers, and control. ELISA was performed on the polyvinyl chloride plates coated with killed whole cell($10^8\;cell/ml$) of S. typhi 0901W by poly-L-lysine applied as binding substance (and polyvinyl chloride as solid phase). The distribution of the level of IgG subclass antibodies in each group was analyzed and compared with other groups. The results obtained were summarized as follow: 1. The optimal dilution of the sera from patients with typhoid fever was 1:160, and those of the sheep anti-human IgG subclass and the peroxidase conjugated rabbit anti-sheep IgG were 1:4000 and 1:5000, respectively. 2. The absorbance levels of IgG subclass in the sera of typhoid fever patients were as follows; a) IgG1 value is $0.439{\pm}0.110$ b) IgG2 value is $0.416{\pm}0.165$ c) IgG3 value is $0.449{\pm}0.145$ d) IgG4 value is $0.525{\pm}0.154$ IgG subclass levels in the sera of typhoid patients were much higher than in control group and patient with paratyphi A or B as well as other infectious diseases. The sensitivity and the specificity in differential diagnosis of typhoid fever and other febrile diseases were 92% and 79% in the assay of IgG1 respectively, whereas those in the assay of IgG2 were 97% and 72%, respectively (above absorbance 0.3). 3. The absorbance levels of IgG subclass in the serial sera of typhiod fever patients tend to decrease to the level of absorbance 0.3 in 10 months from the onset of illness. 4. The order of absorbance levels of IgG subclass in the serum of each group were typhoid fever, paratyphi A or B, other infectious diseases, control and cancer. 5. For the serodiagnosis of typhoid fever against other febrile diseases, the sensitivity and the specificity in the assay of IgG2 activity were 76% and 93% in absorbance 0.4, respectively. 6. In the distribution of the level of each IgG subclass in the sera of FUO patients which were suspected of typhoid fever, the positive rate was ranged from 36% to 82%. This suggest that more than 50% of FUO patients are caused by S. typhi.
김일한,최은경,하성환,박찬일,차창룡,Kim, Il-Han,Choi, Eun-Kyung,Ha, Sung-Whan,Park, Charn-Il,Cha, Chang-Yong The Korean Society for Radiation Oncology 1988 Radiation Oncology Journal Vol.6 No.1
방사선 조사 후 발생한 잠재치사손상의 회복(PLDR)에 있어 조사선량 및 시간에 따른 환경변화가 회복의 동적양상에 미치는 영향을 Vero 세포계를 이용하여 실험하였다. 배양액을 교환시키지 않고 배양하여 평형기에 도달한 세포에 동물실험용 세시움 조사기로 $1Gy\~9Gy$의 감마선을 조사하고 각 조사조건에서 $O\~6$ 및 24시간동안 정치시킨 후 Agarose가 포함된 새로운 배양액에서 배양하였다. 16Gy를 조사한 동종의 세포를 feeder세포로 첨가하여 배양액내의 전 체세포수를 일정하게 한 조건에서 형성된 세포집락수에 따라 세포의 생존을 정하였다. 잠재치사손상의 회복은 $2\~4$시간 정치후에 포화수준에 도달한 빠른 회복이었다. 방사선량이 증가함에 따라 회복속도는 증가하였고, 포화수준의 회복량도 증가하였다. Linear-quadratic mode에 의한 "방사선량-생존분획" 분석 결과 잠채치사손상이 회복됨에 따라 일차 비활성계수 $\alpha$는 급속히 감소하여 $\beta$에 접근하였고 이차 비활성계수 $\beta$는 미미하게 증가하여 PLDR은 $\alpha$로 표시되는 손상에 주로 영향을 주었다. Multitarget model에 따라 분석한 결과 Do는 변화가 없고 Dq가 증가하였다. 세포 생존분획 이 높은 3Gy 이하의 저선량 영역에서 dose modifying factor가 높아 잠재 치사손상의 회복에 의한 영향이 저선량 영역에서 상대적으로 크게 나타났다. Recovery from potentially lethal damage (PLDR) after irradiation was studied in plateau-phase culture of Vero cells in vitro. Unfed plateau-phase cells were irradiated with dose of 1 to 9Gy using Cs-137 irradiator. Cells then were incubated again and left in situ for 0, 1, 2, 3, 4, 5, 6, and 24 hours and then were trypsinized explanted, and subcultured in fresh RPMI-1640 media containing $0.33\%$ agar. Cell survival was measured by colony forming ability. An adequate number of heavily irradiated Vero cells were added as feeder cells to make the total cell number constant in every culture dish. As the postirradiation in situ incubation time increased, surviving fraction increased by PLDR. The rate of PLDR was so rapid that increased surviving fraction reached saturation level at 2 to 4 hours after in situ incubation. As the radiation dose increased, the rate of PLDR fastened and the magnitude of increased surviving fraction at saturation level by PLOR also increased. In analysis of cell survival curve fitted to the linear-quadratic model, the linear inactivation coefficient $(\alpha)$ decreased largely and reached nearly to zero but the quadratic inactivation coefficient $(\beta)$ increased minimally by increment of postirradiation in situ incubation time. So PLDR mainly affected the damage expressed as $\alpha$, In the multitarget model, significant change was not obtained in $D_0\;but\;in D_q$. Therefore, shoulder region in cell survival curve was mainly affected by PLDR and terminal slope was not influenced at all. And dose-modifying factor by PLDR was relatively higher in shoulder region, that is, in low dose area below 3 Gy.