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절편모충(Trichomonas vaginalis)에 대한 단일클론항체의 생산과 그 특성
임미혜,류재숙,안명희,최영길,민득영 한양대학교 의과대학 1999 한양의대 학술지 Vol.19 No.1
Trichomonas vaginalis, mucosal protozoan parasite of the urogenital tract, is responsible for the most common clinically recognized sexually transmitted disease. Surface antigen of parasitic protozoa play the important roles in disease pathogenesis by serving as attachment factors, direct virulence factors, or mediating evasion of the host immune response through direct suppressoin or antigenic variation. In this study seven monoclonal antibodies against plasma membrane of T. vaginalis HY-1 were prepared and were investigated the effector function of them. Seven monoclonal antibodies were produced and their isotypes were Ig G1 (Tv1, Tv7), Ig G2a (Tv4), Ig M(Tv2, Tv3, Tv5, Tv6). All of the seven monoclonal antibodies recognized the determinants located on the surface of live organism by IFAT. Although these surface reactive seven monoclonal antibodies agglutinated T. vaginalis trophozoites, especially MAb Tv1 and Tv4 could agglutinate eight isolates of T. vaginalis. In immunoblot assay, MAb Tv1 & Tv4 detected single peptide with molecular weight of 60 kDa, and Tv7 detected 80 kDa. On the other hand Tv2 and Tv6 detected two (40 kDa & 120 kDa) or more peptides (62 kDa, 64 kDa & 120 kDa), and Tv3 and Tv5 did not bind any pepetide. From these results, MAb Tv1 reacted with 60 kDa, major surface membrane protein of T. vaginalis and agglutinated trichomonads.
겔여과크로마토그라피에 의한 질트리코모나스의 항원분획 분석
임미혜,심재찬,민득영,안명희,류재숙,최영길 한양대학교 의과대학 1993 한양의대 학술지 Vol.13 No.1
This study was aimed to analyze the antigenicity of T. vaginalis using a high performance gel filtration chormatography(HPLC), enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunoelectotransfer blot (EITB). Soluble antigen was prepared using a bead beater. Antiserum was collected from rabbit immunized previously. The results obtained are as follows: the crude antigen of T.vaginalis was separated into 7 peak by HPLC and the fraction 3 revealed the highest reactivity in DLISA. The immunogenic bands of fraction 3 were visualized at 38, 45, 47, 55, 64, 78, 94, 115, 126 and 180 KDa and the bands at 38KDa and 180KDa showed stronger reactivity than that of equivalents of the crude antigen in EITB. In conclusion the fraction 3 of the crude antigen of T. vaginalis obtained by chromatographic separation may have a high antigenicity and proteins ay 38KDa and 180KDa of the fraction 3 may play an important role in antigen-antibody reaction in T. vaginalis infection.
질트리코모나스 (Trichomonas vaginalis)의 항원성 분석에 관한 연구
송준호,임미혜,민득영 한양대학교 의과대학 1991 한양의대 학술지 Vol.11 No.1
Trichomonas vaginalis is a flagellated protozoan parasite which is a major causative organism of vaginitis in women and of urethritis in men. In the present study to elucidate the immune response involving host-parasite interrelationship, antigen analysis of this parasite was carried out using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), enzyme-linked immunoelec-trotransfer blot (EITB) and enzyme-linked immunosorbent assay (ELISA). For the study sera from 21 Trichomonas vaginalis patients, 7 other parasite-infected patients, 2 candidiasis patients, 17 healthy students and 4 young children under 5 years old were subjected and the results obtained were as follows: 1. In SDS-PAGE, sonicated total antigens from 5 strains of T. vaginalis revealed 20-25 protein profiles ranging 14-110 KDa in molecular weight. 2. On EITB probed with sera from trichomoniasis, immunogenic bands was recognized. But in children the bands of 15, 18 and 23KDa were not seen. 4. In high performance gel filtration chromatography of sonicated total antigen of T. vaginalis, 10 peaks were identified as in ranging 2.6-155 KDa. 5. In ELISA using fractions of 10 peaks from high performance gel filtration chromatography as antigens, values of IgG and IgM antibodies of trichomoniasis patients were significantly higher than those of healthy controls (P〈0.001 or P〈0.001-0.002 respectively). In conclusion with results obtained in the present styudy it is assumed that immunogenic bands in 15, 18and 23KDa on SES-PABE may connote the important significance on immune response in T. vaginalis infection.
권장연,임영희,임미혜,류재숙,안명희,민득영 한양대학교 의과대학 1998 한양의대 학술지 Vol.18 No.2
The purification methods such as bead-beater destruction, trichloroacetic acid(TCA) precipitation and 3-[(3-cholamidopropyl) dimethylammonio]-1-propane sulfonate(CHAPS) solubilization of Trichomonas vaginalis antigens on sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and enzyme-linked immunoelectrotransfer blotting (EITB) were evaluated. Total TCA-precipitated proteins and CHAPS-solubilized extract of T. vaginalis possessed common protein bands whose molecular weights were at 28 kDa, 35 kDa, 40 kDa, 50 kDa, 60 kDa, 100 kDa, 120 kDa, 130 kDa, and 160 kDa, and bead-beater treated trichomonal proteins represented some different spectrum of protein bands from TCA- or CHAPS-treated ones in that they didn't show high M.W. protein bands(〉100 kDa) on SDS-PAGE. But similar antigenic bands with the molecular weights of 60 kDa, 110 kDa, 120 kDa and 140 kDa by EITB were identified in TCA-, CHAPS- and bead-beater treated proteins. In conclusion, TCA-precipitation and CHAPS-solubilization were more stable methods to analyse trichomonal proteins than extraction with bead-beater, and the result will be useful for antigen analysis of Trichomonas vaginalis to elucidate a mechanism of pathogenesis in host-parasite relationship.
단백질을 이용한 저열량 식이요법이 성인여성의 신체조성에 미치는 영향
윤미선 ( Mi Sun Yoon ),임미혜 ( Mi Hyea Leem ) 한국미용학회 2007 한국미용학회지 Vol.13 No.2
The main purpose of this study was to investigate how Low-Calorie Diets using protein supplements makes an impact on the women`s body composition. The period of the program was from Apr. 15th until May 30th. The subjects of the study were selected among those who visited Diet Clinic with over 28% in BMI. The 10 persons normally do not exercise but want medications or operations to lose weight. They were instructed to intake 1,200 Kcal per day for 4 weeks, and to replace dinner by protein supplement of 200 Kcal. The results were as followings. Weight, waist measurement, percent body fat, and body fat mass were significantly decreased while lean soft tissue mass, fat free mass and basal metabolic rate remained almost the same in spite of the reduction of daily-calories by 500Kcal. Therefore, it can be said that Low-Calorie Diets using protein is one of the effective diets. Through the survey conducted, it is investigated if there are some differences as per the psychological characteristics of each person by questioning the participants on the degree of moderation in eating in order to know the effects of the diet. As a result, a person who has the high degree of moderation in eating shows better effects of the diet than those who do not. This proves that psychological characteristics are very important in the treatment of obesity as well as the method of a diet. Based on this research, it is considered that safe and various diet programs, which help moderation in eating, should be continued.