폐흡충(Paragonimus Westermani) 감염 마우스에 있어 비장 B 세포표면의 CD23(FcεRII) 항원의 발현

신명헌 梨花女子大學校 醫科大學 醫科學硏究所 1996 EMJ (Ewha medical journal) Vol.19 No.4

목 적: 윤충 감염시 가장 특징적인 면역반응인 혈청내 lgE의 생산에는 CD23(Fc_eRⅡ) 항원의 발현과 밀접한 관계가있을 것으로 생각되나 이에 대해서는 감염 기생충의 종류에 따라 다른 결과를 보이고 있다. 따라서 이 연구에서는 조직 기생충인 폐흡층을 마우스에 감염시킨 후 비장B 세포표면에 CD23(Fc_eRⅡ) 항원의 발현을 측정하여 폐흡충 감염마우스에 있어 증가되는 혈청내 lgE 생산에 CD23 항원의 역할을 알아보고자 하였다. 방 법: 폐흡충의 제2중간숙주인 참가재에서 분리한 폐흡충 피낭유충을 4~6주령의 BALB/c 마우스에 한 마리당 20개씩 경구 감염시킨 후 감염 시기별로(0,2,3,4,6주)로 마우스의 비장으로부터 단핵세포를 분리한 후 B 세포표면의 DB23항원의 발현을 유세포분석기로 측정하였다. 또한 정상 마우스로부터 분리한 비장세포에 폐흡충 피낭유충의 분비-배설물을 첨가한 후 6시간 및 24시간 배양한 후 B 세포표면의 CD23항원의 발현을 측정하였다. 결 과: 폐흡충 감염 마우스의 비장 B 림프구중 CD23 항원 양성 세포율은 감염 2주에 43.4±7.52%로 대조군의 26.1±7.52%에 비해 유희한 수준으로 증가되기 시작하여 감염 3주에도 44.4±2.99%로 대조군의 33.6±2.91에 비해 유의한 수준(p<0.05)으로 증가되었다. 또한 감염 4주 및 6주의 CD23 항원 양성 세포율도 각각 35.7±3.29% 및 38.9±4.96%로 대조군의 30.7±5.94% 및 31.1±3.77%에 비해 증가되었으나 통계적으로 유의한 차이는 보이지 않았다. 정상 마우스의 비장으로부터 분리한 단핵세포에 폐흡충 피낭유충의 분비-배설물을 첨가하였을 때 배양 6시간 후의 B 림프구중 CD23 양성 세포율은 24.8±2.78%로서 배지만을 첨가한 대조군의 18.6±3.01%에 비해 약간 증가되었으나 유의한 차이는 보이지 않았다. 그러나 배양 24시간 후의 CD23 양성세포율은 60.1±7.54%로서 대조군의 17.8±1.54%에 비해 유의한 수준(p<0.05)으로 증가되었다. 결 론:폐흡충 감염 마우스에 있어 혈청내 lgE의 생산은 CD23 발현과 밀접한 관계가 있으며 이때 폐흡충 유충의 분비-배설물이 비장 B 세포표면의 CD23 항원의 발현을 증가시키는 것으로 생각된다. Object : Elevation of serum lgE is the most characteristic immune response in helminthic infections. Recently, expression of CD23(Fc_eRⅡ) on B lymphocytes play a major role in lgE production in allergic diseases. However, the mechanisms causing increased lgE production during helminthic infections are poorly understood. In the present study, the expression of CD 23 on splenic B lymphocytes during the course of infection with paragonimus westermani was examined. Methods : Female, 4-6-week old BALB/c mice were inoculated orally with 20 metacercariae of P.westermani. For detection of CD23 positive B lymphocytes by flow cytometry, splenocytes from infected mice and non-infected age matched controls were stained with FITC-conjugatedrat anti-mouse DB23 and PE-conjugated rat anti-mouse CD45R/B220 mono-clonal antibody. And also, to observe the effect of metacercarial ESP on the expression of CD23 antigen, splenocytes from non-infected mice were incubated in the presence of ESP at 37℃ for 6 h and 24 h. Results : The frequency of CD23 positive B lymphocytes of infected mice was increased significantly(p<0.05) at two and three weeks after infection(43.4±7.52% and 44.4±2.99%, respectively) and persisted the higher levels at four and six weeks after infection. Expression of CD23 antigen of cultured splenocytes from non-infected mice in the presence of metacercarial ESP was increased significantly(p<0.05) at 24 h after incubation(60.1±7.54%). Conclusion : These results suggest that the expression of CD23 antigen induced by metacercarial ESP might play a important role in serum lgE production in mice infected with P.westermani. Object : Elevation of serum IgE is the most characteristic immune response in helminthic infections. Recently, expression of CD23(Fc_eRⅡ) on B lymphocytes play a major role in IgE production in allergic diseases. However, the mechanisms causing increased IgE production during helminthic infection are pooly understood. In the present study, the expression of CD 23 on splenic B lymphocytes during the course of infection with Paragonimus westermani was examined. Methods : Female, 4-6 week old BALB/c mice were inoculated orally with 20 metacercariae pf P. westermani. For detection of CD23 positive B lymphocytes by flow cytometry, splenocytes from infected mice and non-infected age matched controls were strained with FITC-conjugated rat anti-mouse CD23 and PE-conjugated rat anti-mouse CD45R/B220 mono-clonal antibody. And also, to observe the effect of metacercarial ESP on the expression of CD23 antigen, splenocytes from non-infected mice were incubated in the presence of ESP at 37℃ for 6h and 24h. Results : The frequency of CD23 positive B lymphocytes of infected mice was increased significantly(P<0.05) at two and three weeks after infection(43.4±7.52% and 44.4±2.99%, respectively) and persisted the higher levels at four and six weeks after infection. Expression of CD23 antigen of cultured splenocytes from non-infected mice in the presence of metacercarial ESP was increased significantly(p<0.05) at 24h after incubation (60.1±7.54%). Conclusion : These results suggest that the expression of CD23 antigen induced by metacercarial ESP might play a important role in serum IgE production in mice infected with P.westermani.

폐흡충 감염 마우스에 rIFN-γ 처치시 혈청내 IgE 및 말초혈액내 호산구수에 미치는 영향

신명헌 梨花女子大學校 醫科大學 醫科學硏究所 1997 EMJ (Ewha medical journal) Vol.20 No.1

Objective : Elevated serum lgE and pheripheral blood eosinophilia are immunologic hallmark in helminthic infections. Recently, these responses are known to be regulated by Th2-specific cytokine IL-4 and IL-5, respectively. And also, the antagonistic effects of IFN-y on Th2 cell proliferation were shown in vitro. However, few studies on the effect of IFN-y on Th2 cytokine responses in Paragonimus westermani infection are reported, In this study, effects of rIFN-y on serum lgE production and the number of pheripheral blood eosinophils in mice infected with P.westermani were examined. Methods : 5-6week old male BALB/c mice treated with IFN-y were divided into 3 groups. All the mice were inoculated orally with 20 metacercariae of P.westermani. GroupⅠmice(0-14days) were treated intraperitoneally with 2×10^3 unit of rIFN-h at daily intervals from the time of the infection to 4th day infection, group Ⅱ mice(5-14 days) were treated with rIFN-y from the 5th to the 14th day of infection and group Ⅲ mice(8-14 days) were treated from the 8th to the 14th of infection. Total serum lgE and the number of pheripheral blood eosinophils were examined in infected mice treated with rIFN-y. Results : The serum lgE levels in groupⅠand Ⅱ were decreased compared with those of infected mice with no treatment with rIFN-y, but not significantly. The number of pheripheral blood eosinophils in groupⅠand Ⅱ were decreased compared with those of infected mice with no treatment with rIFN-y, especially significant(p<0.05) reduction was shown in groupⅠ. However, the serum lgE levels and number of pheripheral blood eosinophils in group Ⅲ were similar to those of infected mice with no treatment with rIFN-y. Conclusion : These results suggest that IFN-y decreases Th2 cytokine response in P.westermani-infected mice. However, IFN-y treatment has less of an effect once the production of Th2-associated cytokines has become established.

線形스토캐스틱 시스템의 適應的 推定

申明憲,朴民號,金永基 慶北大學校 1980 論文集 Vol.29 No.-

A method for identifying the transition and the control coefficients of a linear discrete-time dynamic system is presented. The transition and control coefficients are identified by requiring that the mean of the measurement residual sequence be zero. The speed of the identification is compared with the Martin and Stubberud's method. And the convexity of the mean value function of the measurement residual sequence is prooved.

ADM 回路 設計에서의 時定數 영향

申明憲 경북대학교 공과대학 1979 工大硏究誌 Vol.8 No.-

The circuitry for CVSD system is designed through digital technique and implemented with CMOS IC's. The system performance is tested experimentally and showed 29dB of SNR for the clock rate of 36Kb/s. The effects of time constants on SNR are experimented in both syllabic and prediction filters.

폐흡충(Paragonimus westermani) 감염백서에서의 혈청내 IgE 항체가의 변동

신명헌(Myeong-Heon Shin),류재숙(Jae-Sook Ryu),민득영(Duk-Young Min) 대한기생충학열대의학회 1991 The Korean Journal of Parasitology Vol.29 No.4

질트리코모나스에 대한 마우스의 세포매개성 면역반응에 관한 연구

신명헌,민득영,류재숙,안명희,신수재 한양대학교 의과대학 1990 한양의대 학술지 Vol.10 No.1

This study was aimed to observed the cell-mediated response by duration after immunization with T. vaginalis in mice. Trichomonas vaginalis, isolated from vulvovaginitis patients, was cultured in the TPS-1 medium axenically. For the experiments trichomonads were inoculated subcutaneously with 1×10?? of trophozoites into the back of BALB/c mice. Following immunization, delayed type hypersensitivity (DTH) response in the footpad and blastogenic response of the mouse spleen cells using [³H]-thymidine were observed on week 1, 4 and 7 after immunization. In the preparation of T. vaginalis lystae, trophozoites wer disrupted with ultrasonicator. Suspension was sedimened, filtered and dialyzed. For the collection of T. vaginalis excretory-secretory product, trophozoites were suspended in RPMI 1640 medium and incubated at 37℃ for 3-4 hours. The supernatant was filtered and dialyzed. T. vaginalis lysate, excretory-secretroy product and PHA were used as mitogens in the blastogenic responses. ELISA technique was applied for the measurement of the serum antibody titer. The results obtained are as follows: 1. The blastogenic response of splenocytes to PHA was slightly decreased on week 4 after immunization, whereas those to T. vaginalis lysate and excretory-secretory product were similar with control group on same time course. Both of these blastogenesis were markedly increased on week 7 after immunization. 2. In regard to delayed type hypersensitivity responses in mice immunized with T. vaginalis, the levels were increased in comparison with the control group throughout 7 weeks after immunization. 3. The serum antibody titers in mice immunized with T. vaginalis were increased continuously from on week 1 to on week 8 after immunization. With these results it is assumed that not only humoral immunity but also cell mediated immunity is involved in immune defense mechanism in the mice experimentally inoculated with Trichomonas vaginalis.

Production of interferon-gamma and interleukin-4 by splenocytes in mice infected with Paragonimus westermani

Myeong-Heon SHIN(신명헌),Duk-Young MIN(민득영) 대한기생충학열대의학회 1996 The Korean Journal of Parasitology Vol.34 No.3

Dermal mast cell responses in Paragonimus westermani-infected mice

Myeong-Heon SHIN(신명헌) 대한기생충학열대의학회 1997 The Korean Journal of Parasitology Vol.35 No.4

Antibody Induced Capping of Surface Antigens in Trichomonas vaginalis

Ryu,Jae-Sook,Shin,Myeong-Heon,Min,Duk-Young INSTITUTE OF TROPICAL MEDICINE YONSEI UNIVERSITY 1992 YONSEI REPORTS ON TROPICAL MEDICINE Vol.23 No.1

기생충의 숙주에 감염시 여러 가지 방법으로 숙주의 면역반응을 회피(escape)한다고 한다. 이질아메바, 트라파노조마, Naegleria fowleri 등에서는 숙주 면역반응의 회피기전의 일환으로 충체 표면항원(surface antigen)을 외부로 탈락시키거나 재생산(renewal)함이 보고된 바 있다. 이 연구에서는 질트리코모나스에서 표면항원의 capping(antigen redistribution)이 일어나는지 관찰하였다. Log-stage의 살아있는 충제(2×10(6))를 질트리고모나스로 면역시킬 토끼혈청(1:5)과 4℃ 30분간 반응시킨 후 fluorescein conjugated goat anit-rabbit IgG(1:30)와 4℃에서 30분 반응시키고 세척 후 즉시 1% paraformaldehyde로 고정하거나 37℃에서 30분내지 2시간 방치한 후 고정하고 형광현미경으로 관찰하였다. 또한 대사억제제인 sodium azide와 미세소관(microtubule)합성 억제제인 vincristine sulfate와 colcemid를 혈청 및 형광항체에 넣어 capping에 영향을 미치는지 관찰하였다. 충체를 4℃ 또는 25℃에서 30분 내지 2시간 반응시킨 군에서는 20% 정도의 copping의 관찰되는데 비해 TSP-1배지에서 37℃, 2시간 반응시켰을 때 60∼70%가 일어났다. 대개 polar capping이었는데, 일부의 충체는 형광물질을 완전히 벗어버려 충체에는 형광이 없었다. Sodium azide의 여러농도(0,025M-0.5M)에서 capping이 현저히 저하되었다. 그러나, vincristine sulfate와 colcemid(5-0.25 g/ml)는 capping에 별 영향을 미치지 않았다. 이상의 결과로 질트리코모나스는 표면으로부터 항체를 제거하여 숙주의 면역반응에 저항하는 것으로 사료되며 이 반응은 반응온도 및 신진대사에 의존되었다. The capping phenomenon, a parasite's escape mechanism from host's immune response by removal of the antibody bound to antigens on parasite's surface membrane, was observed in Trichomonas vaginalis in vitro. An isolated strain of T. vaginalis (HY-9) was incubated with rabbit anti-serum at 4℃, 25℃ and 37℃ for 30 minutes, 1 hour and 1 hours, and then treated with fluorescein labelled anti-rabbit IgG. The cell-capping process was observed obviously under fluorescein microscope, and was temperature dependent. Sodium azide, a metabolic inhibitor, inhibited the capping of T. vaginalis, whereas colchicine and vicristine, the microtubule synthesis inhibitors, did not affect the capping formation. These results suggest that antibody-induced membrane antigen modulation of T. vaginalis may have a role to host's immune response.

Involvement of NOX2-derived ROS in human hepatoma HepG2 cell death induced by Entamoeba histolytica

이영아,신명헌 대한기생충학ㆍ열대의학회 2023 The Korean Journal of Parasitology Vol.61 No.4

Entamoeba histolytica is an enteric tissue-invasive protozoan parasite causing amoebic colitis and liver abscesses in humans. Amoebic contact with host cells activates intracellular signaling pathways that lead to host cell death via generation of caspase-3, calpain, Ca2+ elevation, and reactive oxygen species (ROS). We previously reported that various NADPH oxidases (NOXs) are responsible for ROS-dependent death of various host cells induced by amoeba. In the present study, we investigated the specific NOX isoform involved in ROS-dependent death of hepatocytes induced by amoebas. Co-incubation of hepatoma HepG2 cells with live amoebic trophozoites resulted in remarkably increased DNA fragmentation compared to cells incubated with medium alone. HepG2 cells that adhered to amoebic trophozoites showed strong dichlorodihydrofluorescein diacetate (DCF-DA) fluorescence, suggesting intracellular ROS accumulation within host cells stimulated by amoebic trophozoites. Pretreatment of HepG2 cells with the general NOX inhibitor DPI or NOX2-specific inhibitor GSK 2795039 reduced Entamoeba-induced ROS generation. Similarly, Entamoeba-induced LDH release from HepG2 cells was effectively inhibited by pretreatment with DPI or GSK 2795039. In NOX2-silenced HepG2 cells, Entamoeba-induced LDH release was also significantly inhibited compared with controls. Taken together, the results support an important role of NOX2-derived ROS in hepatocyte death induced by E. histolytica.