반복유산을 경험한 환자에서 임신중 태반항원과 동종항원에 노출된 모체 림프구면역반응은 언제부터 소실되나?

최범채,Choi, Bum-Chae,Hill, Joseph A. 대한생식의학회 1998 Clinical and Experimental Reproductive Medicine Vol.25 No.2

The maintenance of a viable pregnancy has long been viewed as an immunological paradox. The deveolping embryo and trophoblast are immunologically foreign to the maternal immune system due to their maternally inherited genes products and tissue-specific differentiation antigens (Hill & Anderson, 1988). Therefore, speculation has arisen that spontaneous abortion may be caused by impaired maternal immune tolerance to the semiallogenic conceptus (Hill, 1990). Loss of recall antigen has been reported in immunosuppressed transplant recipients and is associated with graft survival (Muluk et al., 1991; Schulik et al., 1994). Progesterone $(10^{-5}M)$ has immunosuppressive capabilities (Szekeres-Bartho et al., 1985). Previous study showed that fertile women, but not women with unexplained recurrent abortion (URA), lose their immune response to recall antigens when pregnant (Bermas & Hill, 1997). Therefore, we hypothesized that immunosuppressive doses of progesterone may affect proliferative response of lymphocytes to trophoblast antigen and alloantigen. Proliferative responses using $^3H$-thymidine ($^3H$-TdR) incorporation of peripheral blood mononuclear cells (PBMCs) to the irradiated allogeneic periperal blood mononuclear cells as alloantigen, trophoblast extract and Flu as recall antigen, and PHA as mitogen were serially checked in 9 women who had experienced unexplained recurrent miscarriage. Progesterone vaginal suppositories (100mg b.i.d; Utrogestan, Organon) beginning 3 days after ovulation were given to 9 women with unexplained RSA who had prior evidence of Th1 immunity to trophoblast. We checked proliferation responses to conception cycle before and after progesterone supplementation once a week through the first 7 weeks of pregnancy. All patients of alloantigen and PHA had a positive proliferation response that occmed in the baseline phase. But 4 out of 9 patients (44.4%) of trophoblast antigen and Flu antigen had a positive proliferative response. The suppression of proliferation response to each antigen were started after proliferative phase and during pregnancy cycles. Our data demonstrated that since in vivo progesterone treated PBMCs suppressed more T-lymphocyte activation and $^3H$-TdR incorporation compare to PBMCs, which are not influenced by progesterone. This data suggested that it might be influenced by immunosuppressive effect of progesterone. In conclusion, progesterone may play an important immunological role in regulating local immune response in the fetal-placental unit. Furthermore, in the 9 women given progesterone during a conception cycle, Only two (22%) repeat pregnancy losses occured in these 9 women despite loss of antigen responsiveness (one chemical pregnancy loss and one loss at 8 weeks of growth which was karyotyped as a Trisomy 4). These finding suggested that pregnancy loss due to fetal aneuploidy is not associated with immunological phenomena.

반복 유산환자의 말초혈액 단핵구와 태반항원을 체외 공동 배양시 세포 매개 면역반응에 프로게스테론이 미치는 영향

최범채,Choi, Bum-Chae,Hill, Joseph A. 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.3

Progesterone is necessary for successful pregnancy and had immunosuppressive properties. Peripheral blood mononuclear cells (PBMC) from many women with unexplained recurrent spontaneous abortion responded to trophoblast extract in vitro by prolifertion and releasing soluble, heat-labile factors that are toxic to mouse embryos (embryotoxic factors). Accumulating evidence suggests that T Helper (Th)-1 type immunity to trophoblast is correlated with embryotoxic factor production and is associated with pregnancy loss, while Th2-type immunity is associated with successful gestation. The objective of this study was to determine whether progesterone can inhibit Th1-type cytokine secretion (IFN-${\gamma}$, TNF-${\alpha}$) by trophoblast-activated peripheral blood mononuclear cells from 23 nonpregnant women (age 25-35) with unexplained recurrent abortion (median 5, range 3 to 15)who otherwise produce embryotoxic factors in response to trophoblast. We also determined whether progesterone affected Th2-type cytokines (IL-4, IL-10) in this system in vitro and if IL-10 (1,500 pg/mL) could inhibit Th1-type immunity to trophoblast. IFN-${\gamma}$ was detected in 17 of 23 (74%) trophoblast stimulated PBMC culture supernatants ($77.94{\pm}23.79$ pg/mL) containing embryotoxic activity. TNF-${\alpha}$ was detected in 19 (83%) of these same supernatants ($703.15{\pm}131.36$ pg/mL). In contrast, none of the supernatants contained detectable levels of IL-4 or IL-10. Progesterone ($10^{-5}$, $10^{-7}$, $10^{-9}$M) inhibited Th1-type immunity in a dose dependent manner, but had no effect on Th2-type cytokine secretion. The inhibitory effects of progesterone were abrogated with RU486, but did not affect Th2-type cytokine secretion in trophoblast-activated cell cultures. IL-10, like progesterone also inhibited Th1-type cytokine secretion but had no effect on Th2-type cytokines. These data suggest that therapies designed to suppress Th1-type cytokine secretion in women with recurrent abortion who have evidence of Th1-type immunity to trophoblast may be efficacious in preventing pregnancy loss and should be tested in appropriately designed clinical trials.

Early Pregnancy Disorders (Recurrent Miscarriage): Pregnancy Success and Failure

최범채 ( Bum Chae Choi ) 대한산부인과학회 2002 교육강연 Vol.2002 No.-

국제기업가정신을 통한 의료기관의 해외진출 전략 : 시엘병원의 몽골진출 사례를 중심으로

최범채(Bum Chae Choi),조세화(Se Hwa Cho),강성호(Seongho Kang) 조선대학교 지식경영연구원 2022 기업과 혁신연구 Vol.45 No.2

Expression of Immunosuppression-Related Genes in Fetal Chorionic Villi Derived from Recurrent Spontaneous Abortion Patients

신주미,김정욱,최범채,이숙환,백광현,Shin, Ju-Mi,Kim, Jeong-Wook,Choi, Bum-Chae,Lee, Sook-Hwan,Baek, Kwang-Hyun The Korean Society for Reproductive Medicine 2003 Clinical and Experimental Reproductive Medicine Vol.30 No.3

연구목적: 본 연구에서는 정상 환자와 습관성 유산 질환 환자에서 유래된 융모막 조직 내에서의 면역억제유전자들의 발현 양상에 대해 알아보고자 하였다. 연구재료 및 방법: 임신 6주와 8주의 습관성 유산 질환 환자와 정상 환자로부터 융모막 조직을 채취하였다 (Normal N=6; RSA N=6). 면역조직화학적 분석을 통해서 조직을 관찰하고 세포가 살아있음을 확인한 후에, 역전사 중합효소연쇄반응을 통해서 면역억제유전자인 placenta protein 14 (PP14), indoleamine 2, 3-dioxygenase (IDO) 그리고 mucin1 (MUC1) 유전자의 발현 정도를 비교하였다. GAPDH 발현에 기준한 면역억제유전자 발현을 정량 분석하여 Student's t-test를 시행하였고, p<0.05를 유의성이 있는 것으로 판정하였다. 결 과: 습관성 유산 질환 환자의 경우, 임신 6주와 8주의 융모막 조직에서의 면역억제유전자 (PP14, IDO, MUC1) 발현이 현저하게 낮은 양상을 보이고 있었다. 습관성 유산과 면역억제유전자의 발현이 통계학적으로 유의성 있는 연관성을 가지고 있다는 것이 확인되었다. 결 론: 면역억제유전자 (PP14, IDO, MUC1)의 발현이 습관성 유산 질환 환자에서 특이적으로 낮게 나타나는 것으로 보아 습관성 유산 질환의 진단과 치료 연구 방안에 이 유전자들의 발현이 이용될 수 있을 것으로 사료된다.

난관수종액이 초기 임신 과정에 태반 세포 증식에 미치는 영향

이지애(Jee Ae Lee),최범채(Bum Chae Choi),변혜경(Hye Gyung Byun),김정욱(Jung Wook Kim),한정렬(Jung Ryul Han),유근재(Geun Jae Yoo),김계현(Kye Hyun Kim),궁미경(Mi Gyung Koong),조셉 힐(Joseph A. Hill) 대한산부인과학회 2000 Obstetrics & Gynecology Science Vol.43 No.8

Objective : To assess the effect of hydrosalpingeal fluid on trophoblast cell proliferation and evaluate the cytokine level in hydrosalpingeal fluid Materials and Methods : Hydrosalpingeal fluid samples obtained from 10 women at the time of laparoscopic surgery. Trophoblast cell proliferation was assayed by a colorimetric immunoassay, based on the measurement of BrdU incorporation during DNA synthesis in response to hydrosalpingeal fluid and the levels of the Th-1 and Th-2 type cytokine determined by sandwich ELISA. Statistical analysis was performed by regression analysis and nonparametric testing(Mann-Whitney U test). Results : Samples from 7 out of 10 patients significantly suppressed trophoblast proliferation in a dose- dependent manner compared to controls(r=-0.0673, p<0.05). IFN-γ and IL-6 were present in 2 out of 10 hydrosalpingeal fluid samples. IL-10 was not detected in all 10 samples. TNF-α, Th-1 type cytokine, was checked in all samples [median(range); 13.12 pg/ml(8.27 pg/ml~19.67 pg/ml)], and the mean concentration of TNF-α was significantly greater in group that suppressed trophoblast proliferation than in group that did not(14.74 pg/ml ± 1.05 pg/ml versus 9.33 pg/ml ± 0.55 pg/ml, p=0.0164). Conclusions : These findings suggest that hydrosalpingeal fluid may negatively affect trophoblast cell proliferation leading to implantation failure and also high level of the Th-1 type cytokine, TNF-α, in hydrosalpingeal fluid may be a potential mechanism for early pregnancy loss.

난소기능평가를 위한 Gonadotropin Releasing Hormone Agonist Stimulation Test (GAST)의 효용성에 관한 연구

김미란,송인옥,연혜정,최범채,백은찬,궁미경,손일표,이진우,강인수,Kim, Mee-Ran,Song, In-Ok,Yeon, Hye-Jeong,Choi, Bum-Chae,Paik, Eun-Chan,Koong, Mi-Kyoung,Song, Il-Pyo,Lee, Jin-Woo,Kang, Inn-Soo 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.2

Objectives: The aims of this study are 1) to determine if GAST is a better indicator in predicting ovarian response to COH compared with patient's age or basal FSH level and 2) to evaluate its role in detecting abnormal ovarian response. Design: Prospective study in 118 patients undergoing IVF-ET using GnRH-a short protocol during May-September 1995. Materials and Methods: After blood sampling for basal FSH and estradiol $(E_2)$ on cycle day two, 0.5ml (0.525mg) GnRH agonist ($Suprefact^{(r)}$, Hoechst) was injected subcutaneously. Serum $E_2$ was measured 24 hours later. Initial $E_2$ difference $({\Delta}E_2)$ was defined as the change in $E_2$ on day 3 over the baseline day 2 value. Sixteen patients with ovarian cyst or single ovary or incorrect blood collection time were excluded from the analysis. The patients were divided into three groups by ${\Delta}E_2$; group A (n=30):${\Delta}E_2$<40 pg/ml, group B (n=52): 40 pg/ml${\leq}{\Delta}E_2$<100 pg/ml, group C (n=20): ${\Delta}E_2{\leq}100$ pg/ml. COH was done by GnRH agonist/HMG/hCG and IVF-ET was followed. Ratio of $E_2$ on day of hCG injection over the number of ampules of gonadotropins used ($E_2hCGday$/Amp) was regarded as ovarian responsiveness. Poor ovarian response and overstimulation were defined as $E_2$ hCGday less than 600 pg/ml and greater than 5000 pg/ml, respectively. Results: Mean age $({\pm}SEM)$ in group A, B and C were $33.7{\pm}0.8^*,\;31.5{\pm}0.6\;and\;30.6{\pm}0.5^*$, respectively ($^*$: p<0.05). Mean basal FSH level of group $A(11.1{\pm}1.1mlU/ml)$ was significantly higher than those of $B(7.4{\pm}0.2mIU/ml)$ and C $(6.8{\pm}0.4mIU/ml)$ (p<0.001). Mean $E_2hCGday$ of group A was significantly lower than those of group B or C, i.e., $1402.1{\pm}187.7pg/ml,\;3153.2{\pm}240.0pg/ml,\;4078.8{\pm}306.4pg/ml$ respectively (p<0.0001). The number of ampules of gonadotropins used in group A was significantly greater than those in group B or C: $38.6{\pm}2.3,\;24.2{\pm}1.1\;and\;18.5{\pm}1.0$ (p<0.0001). The number of oocytes retrieved in group A was significantly smaller than those in group B or C: $6.4{\pm}1.1,\;15.5{\pm}1.1\;and\;18.6{\pm}1.6$, respectively (p<0.0001). By stepwise multiple regression, only ${\Delta}E_2$ showed a significant correlation (r=0.68, p<0.0001) with $E_2HCGday$/Amp, while age or basal FSH level were not significant. Likewise, only ${\Delta}E_2$ correlated significantly with the number of oocytes retrieved (r=0.57, p<0.001). All four patients whose COH was canceled due to poor ovarian response belonged to group A only (Fisher's exact test, p<0.01). Whereas none of 30 patients in group A (0%) had overstimulation, 14 patients among 72 patients (19.4%) in group B and C had overstimulation (Fisher's exact test, p<0.01). Conclusions: These data suggest that initial $E_2$ difference after GAST may be a better prognostic indicator of ovarian response to COH than age or basal FSH level. Since initial $E_2$ difference demonstrates significant association with abnormal ovarian response such as poor ovarian response necessitating cycle cancellation or overstimulation, GAST may be helpful in monitoring and consultation of patients during COH in IVF-ET cycle.

동결수정란 이식주기에서 수정란 융해 후 생존율과 임신율에 영향을 미치는 요인

김정욱,변혜경,염혜원,전진현,박용석,송인옥,송지홍,최범채,궁미경,전종영,강인수,Kim, Jeong-Wook,Byun, Hye-Kyung,Youm, Hye-Won,Jun, Jin-Hyun,Park, Yong-Seog,Song, In-Ok,Song, Ji-Hong,Choi, Bum-Chae,Koong, Mi-Kyoung,Jun, Jong-Young,Kang, 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.1

Objective: The purpose of this study was to determine the important factors affecting survival and pregnancy rate in frozen-thawed embryo transfer cycles. Methods: we performed retrospective analysis in 738 cycles of frozen-thawed embryo transfers, in relation to the insemination methods, the freezing stage of embryo, patient's age, infertility factors and the origin of injected sperm in ICSI cycles. After conventional IVF or ICSI, the supernumerary PN stage zygotes or multicellular embryos were cryopreserved by slow freezing protocol with 1,2-propanediol (PROH) as a cryoprotectant. Results: The survival rates of thawed embryos were 69.3% (1585/2287) in conventional IVF group and 71.7% (1645/2295) in ICSI group. After frozen-thawed embryo transfers, 27.0% (92/341) and 32.0% (109/341) of pregnancy rates were achieved in conventional IVF and ICSI group, respectively. There were no significant difference in the survival and pregnancy rates according to the insemination methods, the freezing stage and patient's age. However, the pregnancy rate (36.2%) of male factor infertility was significantly higher than the tubal (27.2%) and other female factor infertility (22.9%). In ICSI group, the origin of injected sperm did not affect the outcome of frozen-thawed embryo transfer cycles. Conclusion: The present study demonstrates that acceptable clinical outcomes can be achieved after the transfer of frozen-thawed embryos regardless of the stage of embryos for freezing, the patient's age and the origin of injected sperm.

체외수정 - 배아이식 시술후 발생한 양측 난관임신 1 예

김희철,김희철(Hee Chul Kim),최범채(Bum Chae Choi),이재훈(Jae Hoon Lee),양광문(Kwang Moon Yang),홍수정(Soo Jung Hong),김혜선(Hye Sun Kim),김석훈(Seog Hun Kim),김창헌(Chang Heon Kim),강인수(Inn Soo Kang) 대한산부인과학회 2000 Obstetrics & Gynecology Science Vol.43 No.4

Over the last 20 years, the frequency of multiple pregnancy has increased mainly because of the introduction of exogenous pituitary gonadotropins in the treatment of infertility. Since the advent of assisted reproductive technology, the concern about ectopic implantation of embryos has increased dramatically and it continues to be a major complication of in vitro fertilization and embryo transfer (IVF-ET). Bilateral tubal pregnancy is the least common type of ectopic implantation of two embryos. Of all extrauterine pregnancies, 1:725∼1:1580 are bilateral1,2. Simultaneous tubal pregnancies have been reported in natural cycles, recently, after ovulation induction, in-vitro fertilization2,3. The high incidence of ectopic pregnancy associated with IVF-ET continues to be a problem and frequencies of between 2.4 and 12.4% have been only a few reports of simultaneous bilateral tubal pregnancies following IVF-ET4. This paper describes a case of a simultaneous bilateral tubal pregnancy after IVF-ET in a 33 year old patient. The diagnosis was confirmed by diagnostic laparoscopy performed 25 days after embryo transfer(sixth week of gestation), which revealed bilateral tubal pregnancy. Bilateral salpingectomy was performed. With a review of the literature on this topic, diagnostic aspects and treatment options are discussed.

혈청 항 - 카디오리핀 항체 양성인 반복 유산환자의 임신 부산물에 대한 염색체 분석

이동진(Dong Jin Lee),유근재(Keun Jai Yoo),최범채(Bum Chae Choi),김석훈(Suk Hoon Kim),허걸(Hur Kuol),김계현(Kye Hyun Kim),박지혜(Chi Hye Park),박소연(So Yeon Park),최규홍(Kyu Hong Choi),조동희(Dong Hee Cho),강인수(In Soo Kang) 대한산부인과학회 2000 Obstetrics & Gynecology Science Vol.43 No.5

Objective : It has been suggestes that various mechanism of fetal loss are associated with anticardiolipin(ACA) and humoral immunity in the patients with recurrent spontaneous abortion. Thus we have investigated the relationship between ACA and chromosomal anomaly to know the clinical impact of ACA to early fetal loss as comparing to the chromosomal anomaly in the patients of recurrent spontaneous abortions. Materials and Methods : Patients(n=88) with a history of recurrent spontaneous abortion (2 or more) between January 1, 1994 and June 30, 1999 were included in this study. Quantitative measurement of serum ACA was performed by ELISA and chromosomal analysis of chorionic villi obtained from aborted conceptuses was done by using standard G-banding technique. Results: The incidence rate of ACA positive was 27%(24/88) and that of chromosomal abnormality was 57%(50/88). The incidence rate of abnormal karyotype was 54% (13/24) in ACA positive. Among 24 ACA positive, 10 had IgG-ACA positive, 10 had IgM-ACA positive and 4 had both types of ACA. The incidence rate of chromosomal anomalies was 30% (3/10) in IgG-ACA positive, 90%(9/10) in IgM-ACA positive and there was significant difference between these two groups (p=0.02). The incidence rate of chromosomal trisomy was 59% (23/37) in ACA negative, 62% (8/13) in ACA positive and there was no significance between two groups. Conclusion : The significantly low incidence of chromosomal abnormalities in conceptal products of patients with IgG-ACA comparing that of patients with IgM-ACA suggests that this isotype of antibody have influence on the genesis of spontaneous abortions in genetically normal pregnancy. In further studies, additional trials are mandatory for obtaining a definitive conclusions about relationship between pathologic changes of conceptal products and pathophysiologic effects of IgG-ACA.