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항거식세포혈청 및 항임파구혈청이 항체산생 및 서라균(鼠癩菌) 감염에 미치는 영향

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 Effect of anti-macrophage serum (AMS) and anti-lymphocyte serum (ALS) on antibody production and murine leprosy infection in the Korean chipmunks 
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[한글]항거식세포혈청(AMS) 및 항임파구혈청(ALS)의 면역억제작용에 관해서는 많은 학자들의 보고가 있다. 그러나 이들 항혈청의 면역억제작용기전에 관해서는 아직도 명백히 밝혀진 바 없으며 특히 AMS의 면역억제효과에 대해서는 아직도 학자들간에 논란의 대상이 되고있다. 또한 ALS의 면역억제작용에 있어서도 세포성면역의 억제는 여러 연구자들에 의하여 입증되고 있으나 체액항체면역의 억제에 관해서는 항원의 종류나 실험동물의 유전적 배경에 따라 면역억제효과에 차이가 있음을 보고하고 있다. 최근 몇몇 연구자들은 ALS가 실험동물에 있어서 세포내기생세균의 감염을 증강시킬 수 있음을 보고하였다. 이에 저자는 한국산 다람쥐에 있어서 AMS 및 ALS의 투여가 장티프스 백신에 대한 항체산생에 미치는 영향과 서라균의 감염에 미치는 영향을 추구하기 위하여 본 실험을 실시한 바 다음과 같은 결과를 얻었다. 1. 항거식세포혈청 및 항임파구혈청의 투여는 장티프스 백신에 대한 항체산생을 현저히 억제하였다. 2. 항거식세포혈청 및 항임파구혈청의 항체산생억제효과는 시간이 경과함에 따라 서서히 감퇴되는 것으로 사료되었다. 3. 서라균의 복강내 접종에서는 대조군에 비하여 항거식세포혈청 및 항임파구혈청 투여군에서 접종후 8주까지는 간장 및 비장에서 균의 증식양상을 관찰할 수 있었으나 12주부터는 점차 균이 소실되었다. 한편 병리조직학적 소견은 각군에서 공히 간장 및 비장에 균접종 2주 및 16주에서 거의 육아종 형성을 관찰할 수 없었으나 4주에서 12주 사이에서는 육아종 형성을 관찰할 수 있었다. 4. 서라균의 피하 및 족저력 접종군에서는 대조군에 비하여 항거식 세포혈청 및 항임파구혈청 투여군에서 균접종 8주까지 접종부위에서만 군의 증식양상을 관찰할 수 있었으며 타장기에서는 균을 검출 할 수 없었다. 한편 병리조직학적 소견은 각군에서 공히 접종부위에 균접종후 2주 및 16주에서는 육아종형성은 볼 수 없이 다만 서라균 감염만을 관찰할 수 있었으며 균접종후 4주에서 12주 사이에서는 육아종형성과 함께 서라균의 감염을 관찰할 수 있었다. 이상의 결과로 보아 한국산 다람쥐에 있어서 항거식세포혈청 및 항임파구혈청의 투여는 항체산생을 억제할 수 있으며 서라군의 감염증식을 증강시킬 수 있지만 지속적이며 전신적인 감염에 뚜렷한 영향을 끼치지 못하는 것으로 사료된다.
[영문]Heterologous antisera to different cells have been successfully used for a variety of purposes for many years. Antisera against polymorphonuclear leukocytes, mast cells, lymphocytes and macrophages have been used in recent years to elucidate the roles of these cells in various host responses. Anti-macrophage serum(AMS) and anti-lymphocyte serum(ALS) depress a variety of immune responses. Unanue (1968) prepared AMS and evaluated the properties and possible use of AMS, and thereafter AMS has been widely used for studying the role of macrophages in immunological responses of host (Panijel and Cayeux, 1968; Argyris and Plotkin, 1969; Loewi et al., 1969; Gallily, 1971; Isa, 1971). Unanue (1968), Loewi et al (1969), and Gallily et al (1971) reported that AMS could be cytotoxic against peritoneal macrophages and inhibited the phagocytic activity of peritoneal macrophages, but was not effective for immunosuppression. However, many investigators (Panijel and Cayeux, 1969; Argyris and Plotkin, 1969; Dyminski and Argyris, 1969; Lee and Choi, 1973) made observation that AMS could depress the antibody formation and prolong the skin allograft. Though immunosuppressive effect of AMS are still controversial, the immunosuppressive effect of ALS, especially in cell-mediated immune responses, has been rather well established, and widely applied in the animal experiments and human organ transplantation (Kashiwagi et al, 1968; Allison, 1970). Haman leprosy is caused by Mycobacterium leprae but the organism has never been grown in vitro and only since 1960 the infection has been transmitted to a few animals (Shepard, 1960). Unfortunately, the experimental infection of Mycobacterium leprae into the foot pads of mice and infection in other rodents was mild and remained limited and the lesion was microscopic. To substitute this limited infections of Mycobcterium leprae in mice, efforts have been made to develope more susceptible experimental animals and other immunosuppression methods in order to increase the susceptibility of host animals to Mycobacterium leprae. Lew et al (1974) reported that the Korean chipmunks were susceptible to Mycobacterium leprae and could be used as an experimental animal for leprosy study. More recently it has been shown that the limiting factor has an immunological basis, for a progressive and spreading infection can be produced in mice with a reduced immunological capacity(Rees, 1966; Gangas, 1967, 1968; Gangas and Rees, 1968; Fieldsteel and McIntosh, 1971). These findings led to a surge of interest in the use of immunosupressive agents for the transmission of experimental leprosy in animals. However, several immunosuppresants, such as cortison and suramin, which have been shown to enhance significantly the infection with other species of Mycobacteria, failed to enhance leprosy in mice (Rees, 1964; Shepard et al., 1965). The present experiments were undertaken in order to define the immunosuppressive effect of AMS and ALS on antibody formation and the enhancing effect of AMS and ALS on murine leprosy infection in Korean chipmunks. Material and Methods Preparation of anti-macrophage serum (AMS) and antilymphocyte serum (ALS): AMS and ALS were prepared by 4 times of intramuscular injection of cell mixture with Freund's incomplete adjuvant into rabbits at one week interval, and one week later cell suspensions alone were injected intravenously as a final immunization. A total number of cells inoculated per rabbit was 1.3X10**7 macrophages and 2.4X10**8 lymphocytes. Rabbit anti-chipmunk lymphocyte and macrophage serua were collected 7 days after the last injection of cells and decomplemented and absorbed with chipmunk red blood cells. Peritoneal macrophage donors were injected intraperitoneally 7 days preveously with 5 ml of thioglycollate broth. Peritoneal macrophages were collected and prepared according to Argyris and Plotkin (1969). Lymphocyte suspension was obtained from spleen tissue which were minced and passed through stainless steel wire mesh, and then washed three times with NCTC 135-heparin solution. Immunization of rabbit with typhoid vaccine: Chipmunks were injected intraperitoneally with 0.3ml of AMS and ALS one day before and after injection of thyphoid vaccine 0.25 ml. AMS and ALS were injected consecutively at one week interval, and thyphoid vaccine was also injected once a week for 4 weeks. Chipmunk sera were collected at each week and the sera from each group of five chipmunks were pooled. Serum antibody titers were determined by tube agglutination test. Effect of AMS and ALS on murine leprosy infection: Chipmunks were inoculated with Mycobacterium lepraemurium (Hawaii strain) suspension (4.2X10**6) into peritoneal cavity, subcutanous tissue, and both hind foot pads one day after intraperitoneal treatment with AMS and ALS. After inoculation of Mycobacterium lepraemurium, AMS and ALs, 0.25ml each, were injected intraperitoneally 2 times at 2 days intervals, and followed by 2 times a week for 4 weeks. Chipmunks inoculated with murine leprosy bacilli were sacrificed at 2nd, 4th, 8th, 12th, 16th week, and bacteriological and histopathological studies were made to follow the outcome of the infection. Summary and Conclusion: The present studies were undertaken in order to define the immunosuppressive effect of AMS and ALs on antibody formation against typhoid vaccine and the effect on murine leprosy infection as a preliminary study before the performance of the experiment about the enhancing effect of AMS and ALS on the human leprosy infection in Korean chipmunks, and the results are summarized as follows; 1. AMS and ALS suppressed significantly the antibody formation against typhoid vaccine in Korean chipmunks. 2. It appeared that the suppressive effect of AMS and ALS on antibody formation was gradually decreased in spite of subsequent injections. 3. Murine leprosy infection into the chipmunks treated with AMS and ALS was enhanced by 8 weeks after challenging with Mycobacterium lepraemurium but the enhancing effect was short-lived, and the progressive and wide spreading infection of Mycobacterium lepraemurium in Korean chipmunks by the treatment of AMS and ALs was not observed.
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