주조직적합항원이 불일치하는 마우스 동종 조혈모세포이식에서 IL-2로 유도된 CD4+CD25+ T세포를 이용한 이식편대숙주병의 억제

현재호,정대철,정낙균,박수정,민우성,김태규,최병옥,김원일,한치화,김학기,Hyun, Jae Ho,Jeong, Dae Chul,Chung, Nak Gyun,Park, Soo Jeong,Min, Woo Sung,Kim, Tai Gyu,Choi, Byung Ock,Kim, Won Il,Han, Chi Wha,Kim, Hack Ki 대한면역학회 2003 Immune Network Vol.3 No.4

Background: In kidney transplantation, donor specific transfusion may induce tolerance as a result of some immune regulatory cells against the graft. In organ transplantation, the immune state arises from a relationship between the immunocompromised graft and the immunocompetent host. However, a reverse immunological situation exists between the graft and the host in hematopoietic stem cell transplantation (HSCT). In addition, early IL-2 injections after an allogeneic murine HSCT have been shown to prevent lethal graft versus host disease (GVHD) due to CD4+ cells. We investigated the induction of the regulatory CD4+CD25+ cells after a transfusion of irradiated recipient cells with IL-2 into a donor. Methods: The splenocytes (SP) were obtained from 6 week-old BALB/c mice ($H-2^d$) and irradiated as a single cell suspension. The donor mice (C3H/He, $H-2^k$) received $5{\times}10^6$ irradiated SP, and 5,000 IU IL-2 injected intraperitoneally on the day prior to HSCT. The CD4+CD25+ cell populations in SP treated C3H/He were analyzed. In order to determine the in vivo effect of CD4+CD25+ cells, the lethally irradiated BALB/c were transplanted with $1{\times}10^7$ donor BM and $5{\times}10^6$ CD4+CD25+ cells. The other recipient mice received either $1{\times}10^7$ donor BM with $5{\times}10^6$ CD4+ CD25- cells or the untreated SP. The survival and GVHD was assessed daily by a clinical scoring system. Results: In the MLR assay, BALB/c SP was used as a stimulator with C3H/He SP, as a responder, with or without treatment. The inhibition of proliferation was $30.0{\pm}13%$ compared to the control. In addition, the MLR with either the CD4+CD25+ or CD4+CD25- cells, which were isolated by MidiMacs, from the C3H/He SP treated with the recipient SP and IL-2 was evaluated. The donor SP treated with the recipient cells and IL-2 contained more CD4+CD25+ cells ($5.4{\pm}1.5%$) than the untreated mice SP ($1.4{\pm}0.3%$)(P<0.01). There was a profound inhibition in the CD4+CD25+ cells ($61.1{\pm}6.1%$), but a marked proliferation in the CD4+CD25- cells ($129.8{\pm}65.2%$). Mice in the CD4+CD25+ group showed low GVHD scores and a slow progression from the post-HSCT day 4 to day 9, but those in the control and CD4+CD25- groups had a high score and rapid progression (P<0.001). The probability of survival was 83.3% in the CD4+CD25+ group until post-HSC day 35 and all mice in the control and CD4+CD25- groups died on post-HSCT day 8 or 9 (P=0.0105). Conclusion: Donor graft engineering with irradiated recipient SP and IL-2 (recipient specific transfusion) can induce abundant regulatory CD4+CD25+ cells to prevent GVHD.

Reversal of serologic, immunologic, and histologic dysfunction in mice with systemic lupus erythematosus by long‐term serial adipose tissue–derived mesenchymal stem cell transplantation

Choi, Eun Wha,Shin, Il Seob,Park, So Young,Park, Ji Hyun,Kim, Jong Sung,Yoon, Eun Ji,Kang, Sung Keun,Ra, Jeong Chan,Hong, Sung Hwa Wiley Subscription Services, Inc., A Wiley Company 2012 Vol.64 No.1

<P><B>Abstract</B></P><P><B>Objective</B></P><P>To investigate the efficacy of human adipose tissue–derived mesenchymal stem cell (AD‐MSC) transplantation in systemic lupus erythematosus (SLE) and to determine the optimal transplantation window for stem cells either before or after disease onset.</P><P><B>Methods</B></P><P>(NZB × NZW)F<SUB>1</SUB> mice with SLE were administered human AD‐MSCs (5 × 10<SUP>5</SUP>) intravenously every 2 weeks from age 6 weeks until age 60 weeks, while the control group received saline vehicle on the same schedule. Another experiment was carried out with a different initiation time point for serial transplantation (age 6 weeks or age 32 weeks).</P><P><B>Results</B></P><P>Long‐term serial administration (total of 28 times) of human AD‐MSCs ameliorated SLE without any adverse effects. Compared with the control group, the human AD‐MSC–treated group had a significantly higher survival rate with improvement of histologic and serologic abnormalities and immunologic function, and also had a decreased incidence of proteinuria. Anti–double‐stranded DNA antibodies and blood urea nitrogen levels decreased significantly with transplantation of human AD‐MSCs, and serum levels of granulocyte–macrophage colony‐stimulating factor, interleukin‐4 (IL‐4), and IL‐10 increased significantly. A significant increase in the proportion of CD4+FoxP3+ cells and a marked restoration of capacity for cytokine production were observed in spleens from the human AD‐MSC–treated group. In the second experiment, an early stage treatment group showed better results (higher survival rates and lower incidence of proteinuria) than an advanced stage treatment group.</P><P><B>Conclusion</B></P><P>Serial human AD‐MSC transplantation had beneficial effects in the treatment of SLE, without adverse effects. Transplantation of human AD‐MSCs before disease onset was preferable for amelioration of SLE and restoration of immune homeostasis.</P>

백혈병 마우스 모델의 동종골수이식에서 활성화된 자연살해세포들의 보충이 이식편대백혈병효과와 이식편대숙주반응에 미치는 영향

엄현석,한치화,박수정,김소연,정낙균,정대철,진종률,최일봉,양형모,서영훈,송현근,최인표,민우성,김춘추 대한조혈모세포이식학회 2001 대한조혈모세포이식학회지 Vol.6 No.1

배경: 백혈병에서 동종골수이식 (allogeneic bone marrow transplantation)의 성공적 치료 효과를 얻기 위해서는 이식편대숙주반응 (graft-versus-host disease, GVHD) 발생의 극복과 재발의 방지가 중요한 과제이다. 골수를 역류원심성 세포분리 (counterflow centrifugal elutriation, CCE) 방법으로 분리하여 얻은 rotor off (R/O) 세포분획은 T 세포의 수는 적지만 조혈모세포들을 다량 포함하고 있어 동종골수이식에서 주조직적합복합체 (major histocompatibility complex, MHC) 차이를 극복할 수 있고, 이식편의 생착 성공과 GVHD 발생 예방에 효과적이다. 그러나 골수로부터 T 세포를 제거하면 백혈병세포를 공격하는 이식편대백혈병 (graft-versus-leukemia, GVL) 효과가 감소되기 때문에 백혈병 재발의 빈도가 높다. 자연살해세포 (natural killer cell, NK cell)의 보충 첨가는 동종골수이식 후 GVHD 발생을 줄이면서 충분한 GVL 효과를 얻을 수가 있다. 따라서 저자는 분리 후 IL-2로 활성화시킨 NK 세포들을 골수 R/O 세포분획과 함께 백혈병 마우스 모델에 동종이식함으로써 GVHD와 GVL에 미치는 효과를 관찰하였다. 방법: Balb/c (H-2^(d)) 마우스에서 유래된 A20 (murine B-lymphoma/leukemia cell line, H-2^(d)) 백혈병 세포를 이식 2 일 전에 Balb/c 마우스에 주입하고, 치사량의 전신 방사선을 조사한 직후에 Balb/c 또는 C57BL/6 (H-2^(b)) 마우스의 골수 R/O 세포분획을 꼬리정맥을 통하여 주입하였다. 이들은 모두 이식 후 6-8 주 이내에 사망하였다. 동종이식의 대조군 (n=9)에는 1 × 10^(7)의 R/O 세포분획만을 주입하였고, 실험군 (n=9)에는 C57BL/6 마우스의 비장세포들로부터 단클론항체들을 이용한 negative selection방법으로 분리한 후 IL-2로 활성화된 5 × 10^(5)의 NK 세포분획을 1 × 10^(7)의 R/O 세포분획과 함께 주입하였다. GVL 효과의 판정은 이식 후 14 일과 28 일 째 되는 날 마우스에서 골수, 비장, 간 등을 얻어 백혈병 세포들의 침윤을 조직학적으로 관찰하였으며, GVHD의 정도는 육안적 관찰법으로 평가하였다. 결과: R/O 세포분획만을 이식한 대조군의 골수, 비장, 그리고 간 조직에서는 A20 백혈병 세포의 침윤이 각각 89% (8/9), 78% (7/9)와 22% (2/9)에서 관찰되었고, R/O 세포분획과 NK 세포분획을 함께 이식한 실험군에서는 비장과 간을 제외한 골수에서만 89% (8/9)에서 A20 백혈병 세포의 침윤이 관찰되어 두 군 사이에 장기별 분포의 차이를 볼 수 있었다 (P= 0.0001). 한편 GVHD는 두 군 모두에서 경하게 나타나서 유의한 차이는 없었다. 또한 생착 부전으로 사망한 마우스는 없었다. 결론: CCE를 이용하여 T 세포를 제거한 동종골수이식에서 NK 세포의 보충은 GVHD의 악화는 일으키지 않으면서, 백혈병의 진행을 억제하는 GVL 효과를 얻을 수 있었다. Background: Allogeneic bone marrow transplantation (BMT) with T cell-depleted marrow accompanies engraftment failure and relapse of leukemia by a loss of the graft-versus-leukemia (GVL) effect frequently, while it can prevent GVHD. Supplement of NK cells could prevent GVHD and enhance GVL effect in several murine allogeneis BMT models Roter off (R/O) cell fraction obtained by counterflow centriation elutriatio (CCO) contains small number of T cells and many hematopoietic stem cells. The aim of this study was to determine the effect of R/O cell fraction supplemented with IL-2 activated NK cells on GVL and GVHD within the leukemic mouse BMT model. Methods: Inoculation of A20 (H-2d, murine B-lymphoma/leukemia, Balb/c origin) cells into Balb/c mice via the tail vein 2 days prior to lethal total body irradiation (TBI) and infusion of the Balb/c BM or C57BL/6 (H-2b) R/O fraction were performed. It resulted in 100% mortality within 6 to 8 weeks. The irradiated mice in the control group were injected with 1 × 107 R/O cell fraction alone (n=9) and in the experimental group mice were injected with 1 × 107 R/O cell fraction plus 5 × 105 negatively selected IL-2 activated NK cell fractions of the spleens via the tail vein (n=9). On day 14 and 28 after BMT, the bone marrows, spleens, and livers of mice were harvested for histopathologic analysis of the infiltrations of leukemic cells. We then evaluated the GVHD within the mice. Results: A histopathologic study of the recipients receiving R/O fraction alone showed infiltration of leukemic cells, 89% (8/9) in bone marrows, 78% (7/8) in spleens, and 22% (2/9) in livers. The experimental group of mice showed only the infiltration of leukemic cells 89% (8/9) in bone marrows, not in spleens and livers. There were the organ differences of the leukemic cells infiltrations between the two groups (P=0.0001). There were no obvious differences in the GVHD scores between these two groups, and severe GVHD was not observed. There was no engraftment failure among groups. Conclusion: Thus, our findings suggest that R/O cell fraction obtained by CCE and supplemented with NK cells can promote GVL effect without mediating clinically overt GVHD in allogeneic BMT of mouse leukemia.

막분리법을 이용한 汚水의 再利容에 관한 硏究

박경환,성일화 京畿專門大學 1997 京畿專門大學 論文集 Vol.- No.25

갑상선결절의 임상 및 초음차소견의 진단적 가치

박거운,박철진,박일구,신지혜,정중화,배학연,김상용 朝鮮大學校 附設 醫學硏究所 2007 The Medical Journal of Chosun University Vol.32 No.2

Backgrounds: Thyroid nodule have been relatively common disease and it's prevalence estimated about 4-7%. Recently, high resolution ultrasonography has made the detection of small thyroid nodule possible, Increases in the detection of thyroid nodule have created a clinical dilemma on how to properly manage such thyroid nodules. We investigated the prevalence, clinical and ultrasonographic characterristics, and optimal diagnostic approach toward detected benign and malignant thyroid nodules. Methods: A retrosepctive review was undertaken on the 372 patients who was done Ultrasonograpy guided fine needle aspiration biopsy in Chosun University Hospital, Gwang-Ju, Korea between July 2003 and Spring 2005. Devided into four group such as benign and malignant thyroid nodule on the basis of size 1.5Cm. The review consisted of thyroid function test, antithyroid antibodies, thyroid autoantibodies, thyroid ultrasonography, fine needle aspiration biopsy in each group. Results: The prevalence of thyroid nodule malignancy rate was 18.0%. The prevalence of thyroid incidentaloma malignanct rate was 17.5%. There were no significant differences in age, sex, thyroid function test and size between the benign, malignant incidentalomas and nodule. Ultrasonographic characteristics, include ultrasonography index point showed meaningful diagnostic value for the detection of malignancy in thyroid nodule, Conclusion: Thyroid cancers are fairly common finding. There are no clinical difference between benign and malignant thyroid nodule; however, ultrasonographic findings can be used to decision of optimal management strategies 배경: 최근갑상선 초음파의 빈번한 시행으로 갑상선 결절의 발견이 증가하고 있다. 이러한 갑상선 결절중 특히 갑상션우연종을 임상적으로 어떻게 접근하고 치료를해야하는지는 적지 않는 문제가 되고 있다. 본 연구에서는 갑상선 우연종 및 직경 1.5 cm 이상의 갑상션결절의 유병률, 임상적 특징, 초음파특정 및 초음파의 진단적 유용성에 대해 알아 보았다. 방법: 2003년 7윌부터 2005년 3월까지 조선 대학교병원 내과 및 일반외과및 건강검진 센터를 통해 갑상선 초음파, 경동맥 초음파 및 경부 CT등을 통해 우연히 발견된 갑상션결절을 가진 372명을 대상으로 후향적 연구를 시행하였다. 크기 1.5 cm 미만의 양성및 악성 갑상선 결절군과 크기 1.5 cm 이상의 양성 및 악성결절로 그룹을 나누고 각 군에서 나이, 성별, 갑상선 기능검사, 항갑상선 항체, 갑상선 초음파 및 미세침흡인세포검사등을 검토하였고, 모든 갑상선 결절에 Koike (6)가 제한한 초음파지표 점수(Ultrasonographie index point)를 부여하여 각 군을 비교하였다. 결과: 분석한 갑상선 결절 372예 중 악성결절의 유병율은 18.0% (67예)였다. 갑상선 우연 종의 유병율은 81.5% (303예)였다. 갑상선 우연 종에서의 악성률은 17.5% (53예)로 나왔다. 크기 1.5 cm 이상의 양성 및 악성 갑상선 결절과 양성 및 악성 갑상선 우연종에서 나이, 성별, 갑상선 기능검사, 항갑상선항체에는 의미있는 차이가 없었다. 또한 양성 및 악성우연종에서 의 갑상선 결절크기에도 의미있는 차이는 보이지 않았다. 갑상선 결절의 초음파 지표점수만이 양성 및 악성우연종과 양성 및 악성갑상선 결절의 감별에 의미있는 차이를 보였다. 갑상 선우연종과 크기 1.5 cm 이상의 갑상선 결절에 서 양성을 나타내는 초음파지표점수는 각각 3 점과 4점으로 다르게 나타났다. 양성 및 악성 갑상선우연종의 감별에서 초음파 지표점수의 민감도는 93.6%, 특이도는 52.8%, 양성예측율은 92.4%로 나왔고, 진단적 효율은 86.4% 였으며 크기 1.5 cm 이상의 양성 및 악성 갑상선 결정의 감별에서 초음파지수 민감도는 92.7%, 특이도가 42.9%, 양성 예측도 86%, 효율 82.6%를 보였다. 결론: 갑상선결절에서의 양성 및 악성의 유무는 임상적으로 판단하기 힘들지만 갑상선 결절에서의 초음파 소견 및 초음파 지표 점수를 도입하면 이에 대한 감별에 도움을 줄 수 있다. 따라서 갑상선 초음파검사시 초음파지표점수의 사용은 향후 갑상선 결절의 진단 방향을 제시하는 중요한 길잡이가 될 수 있으리라 사료된다.

폐기물 센서스 기법에 관한 연구

박경환,성일화,민달기 京畿專門大學 1995 京畿專門大學 論文集 Vol.- No.23

인슐린 자가면역 증후군 2예

박일구,박철진,신지혜,김진화,김상용,배학연 朝鮮大學校 附設 醫學硏究所 2008 The Medical Journal of Chosun University Vol.33 No.1

인슐린 자가면역 증후군은 항체에 결합되어 있던 인슐린이 혈당정도와 관계없이 유리됨으로써 저혈당이 야기되는 질환으로, 인슐린종 등과 감별을 필요로 하는 질환이다. 저자들은 고인슐린혈증과 인슐린 자가항체가 양성이면서 공복검사상에서 저혈당이 유발되며 경구 포도당 부하 검사에서 저혈당이 유발된 자가면역성 저혈당증을 경험하였기에 문헌고찰과 함께 보고하는 바이다. Autoimmune hypoglycemia is one of the rare causes of hypoglycemia, and characterized by hyperinsulinemia. Insulin autoimmune syndrome is characterized by insulin autoantibody, hyperinsulinemia, and fasting hypoglycemia without previous insulin immunization. Insulin autoimmune syndrome is usually related to autoimmune disease such as Graves' disease and to previous exposure to drugs. This syndrome is a self-limited disorder. The hypoglycemia was treated with prednisolone and the patient recovered from this. Recently we experinenced two cases of insulin autoimmune syndrome that developed symptomatic hypoglycemia during both the fasting and oral glucose tolerance test. Here in, We present those cases with a review of the literature.

불임여성의 식생활 특성

오지수,정혜자,권혁찬,박원일,궁미경,김화영,Oh, Ji-Soo,Jung, Hye-Ja,Kwon, Hyuck-Chan,Park, Won-Il,Koong, Mi-Kyoung,Kim, Wha-Young 대한생식의학회 2003 Clinical and Experimental Reproductive Medicine Vol.30 No.4

Objective: The purpose of this study was to identify dietary factors related to infertility in Korean women through a case-control study. Methods: The case group was composed of 236 women who had been diagnosed as infertility in hospital. The control group of 181 healthy women with children were recruited from local immunization centers. Socio-economic status, medical history, dietary intakes using food frequency questionnaire and stress were surveyed by interview. Anthropometric measurements were made and the causes of infertility were identified through medical records. Fasting blood samples were taken from subgroup of the subjects. Results: The mean age of infertile and control groups was 31.1 and 32.4 years, respectively and the difference was statistically significant. The mean Body Mass Index of infertile women was not significantly different from control women, however, Waist/ Hip Ratio and Triceps Skinfolds Thickness were significantly lower in infertile women than in control women. The dietary intake status was generally satisfactory in both groups. The intakes of energy, protein, fat, carbohydrate, retinol, vitamin B2 and niacin were lower in infertile women than in control women. The infertile women also showed lower intakes of animal foods. No differences were found between two groups in serum concentrations of albumin, hemoglobin, Fe, TIBC, total cholesterol, HDL-cholesterol, LDL-cholesterol, triglyceride, C3, IgA, IL-2, however, infertile women showed higher levels of Zn and IgG. The stress score was higher in infertile women. Conclusions: From the results of this study, dietary factors and nutritional status do not seem to be directly related to infertility. However, the intertile women have lower nutrient intake and lower body fat content than control women. Further researches are needed according to the causes of infertility for long term to establish the relationship between dietary factors and infertility.

대장에 발생한 낭성 림프관종 1예

박영희,김화영,김재일,박석준,이창후,최동환,유인환,어중건,전동하,박주일,강정협,한성민 대한소화기내시경학회 1996 Clinical Endoscopy Vol.16 No.5

Lymphangioma is a benign tumor of lymphatic origin. Lymphangioma can occur anywhere in the body and only rarely affects the intestinal tract. Most intestinal lymphangiomas are asymptomatic and detected incidentally at autopsy or surgery. Occasionally, they may be large enough to present as a mass to cause obstruction or intussusception. Recently, we examined a case of a 27-year-old woman who complained right upper quadrant abdominal pain, increased bowel sound and weight loss. By surgical resection after the double contrast barium enema and colonoscopy, we could confirm cystic lymphangioma of the transverse colon. So we report this case with brief review of relevant literature.

Purification and Characterization of Arylphorin of the Chinese Oak Silkmoth, Antheraea pernyi

Park, Snag-Bong,Kim, Jeong-Wha,Kim, Soohyun,Park, Nam-Sook,Jin, Byung-Rae,Hwang, Jae-Sam,Seong, Su-Il,Lee, Bong-Hee,Park, Eunju Korean Society of Sericultural Science 2003 International Journal of Industrial Entomology Vol.6 No.1

The arylphorin was purified from the pupal haemolymph of the Chinese oak silkmoth, Antheraea pernyi, and characterized physiologically and biochemically, The protein was purified by a simple preparative polyacrylamide gel electrophoresis (PAGE) and subsequent diffusive elution. The preparation was shown to be homogeneous by 7.5% native-PAGE. The native molecular weight of arylphorin was 450 kDa with a 80 kDa single subunit, suggesting hexamer, The protein contained high amounts (18.3%) of aromatic amino acids, phenylalanine (9.7%) and tyrosine (8.6%). Therefore, the protein was identified as a kind of a storage protein referred to as an arylphorin. The protein was stained by Schiff's reagent, suggesting a glycoprotein. The protein contained 4.9% (w/w) sugar and mannose and N-acetylglucosamine were major components. Also, degradation of the protein was begun by heat treatment at 90 for 20 minutes. These results showed that the A. pernyi arylphorin in the study is hexamer associated with the six subunits consisting of a 80kDa single subunit, and is different from that of Kajiura et al. (1998) in the subunit composition.