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시험관내 복강거식세포에 탐식시킨 서라균의 생체내 증식에 관한 연구

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 Multiplication of macrophage associated mycobacterium lepraemurium in mouse footpad 
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[한글] 거식세포는 체외에서 들어온 항원 또는 세균에 대하여 비특이적 탐식 및 식균작용이 있 고 또한 임파구에 항원정보를 전달하여 특히 면역반응을 유발할 뿐만 아니라 면역과정에 서 활성화된 최종작동세포로 작용함으로써 미생물 감염에 있어 체액항체성 및 세포성면역 기전에 의한 방어작용을 나타낸다. 면역과정에서 거식세포의 역할을 규명하기 위하여 많은 학자들이 연구하였던 바, 일반 항원을 거식세포에 전처치한 후 생체내 투여할 경우 항원만 단독으로 투여한 경우에 비하 여 숙주의 면역반응이 증강(carver 및 Campbell, 1957; Askonas 및 Rhodes, 1965) 혹은 감소(Morita 및 Perkins, 1965; Parkins 및 Makinodan, 1965)한다고 하였으며 한편 Argyr is(1967)는 두 경우 사이에 특별한 차이가 없음을 보고하였다. 이처럼 시험관 내에서 거식세포에 탐식시킨 항원을 생체내 투여할 경우 이 항원에 대한 숙주의 면역반응에 미치는 영향은 아직도 논란의 여지가 많다. 따라서 본 연구에서는, 전술한 보고자들이 실험에 사용한 일반항원들과는 달리, 살아서 증식할 수 있는 항원인 서라균을 이의 숙주세포인 동시에 이 균에 대한 숙주의 주 방어 기전에 있어 최종작동세포로 작용하는 양면성을 지닌 거식세포에, 미리 시험관내에서 탐 식시킨 후 이를 마우스의 족저부에 접종하여 서라균을 단독으로 접종한 대조군과 함께 균 접종 족저부 및 각 장기에서의 서라균 증식양상 및 서라균항원에 대한 지연형 과민반응 출현을 비교 관찰하였던 바 다음과 같은 결과를 얻었다. 서라균을 동계마우스(C^^3H)의 복강거식세포에 탐식시켜 족저부에 접종한 실험군에서 서라균의 장기내 출현시기는 서라균을 단독 접종한 대조군에 비하여 조기에 출현하는 경 향을 보였으나 서라균 접종 족저부에서의 균 증식 및 서라균항원에 대한 지연형 과민반응 에 있어서는 양군간에 차이를 관찰할 수 없었다. 따라서 일반항원을 거식세포에 전처치하 여 투여할 때 관찰할 수 있는 면역반응의 증강 또는 감소와는 달리, 거식세포에 탐식시킨 서라균을 접종했을 때 서라균의 감염에 대한 숙주 방어기전에 영향을 미치지 못하는 것 으로 사료된다. 한편 서라균 10**5 감염후 서라균항원에 대한 지연형 과민반응이 20주 후에 점차 소실 되어감을 관찰할 수 있었으며 이는 서라감염이 진행될수록 세포성면역이 저하되기 때문으 로 사료된다.
[영문] It has been well known that macrophages are taking an important role in the nonspecific host resistance by phagocytizing pathogenic microorganisms and on the other hand, play an essential part in variety of immune precesses including presentation of antigens to lymphocyte, sensitization of lymphocyte, and are acting as activated effector cells. The nature of antigen processing by macrophages has received much attention in recent years. Some investigators reported that immunogenecity of antigens was enhanced by macrophage ingestion (Garvey and Campbell, 1957; Askonas and Rhodes, 1965; Uhr and weissman, 1965). However, others suggested that immunogenic activity of antigens was destroyed by macrophage contact(Morita and Pertains. 1965; Perkins and Makinodan, 1965). On the other hand, Argyris (1967) reported that it was not altered by macrophage contact, that is there was no enhancement or suppression in immunogenecity of antigen by pre-treatment with macrophages in vitro. Thus, the living bacteria which can parasitize and multiply in macrophages were used as a source of antigen instead of non-viable antigenic materials which had been used for these type of researches by others. The living antigen selected was Mycobacteria lepraemurium which is known to he a obligatory intracellular parasite to macrophage, It is the generally accepted fact that the macroptages are the most effective final effector cells in host defense upon mycobacterial infections. This unique relationship may pause a certain contradictory interaction between offensive role of the bacteria and defensive roles of the macrophages in the host. In this experiment, bacteria engulfed macrophages were inoculated into the footpad of the syngeneic mice (experimental group). Bacterial growth in various organs inclunding the footpad of the experimental animals was comparatively studied. Delayed type of hypersensitivity was also examined comparatively with that of the animals inoculated with the bacteria only without any kinds of pre-treatment (control group). The results of this experiment may be summarized as follows; 1. The phagocytic index (number of M. lepraemurium phagocytized by 100 macrophages) reached its maximum value after elapse of 6 hours after inoculation of 0.1ml of M. lepraemurium suspension(1.0×10**8/ml) into the macrophage monolayer(1.7×10**6 macrophages/Leighton tube). 2. A significant increase(10**3 fold) in M. lepraemurium population in mouse footpad was observed at the 12th week after inoculation with 10**5 bacteria into the footpad cia macrophages. However, this increase was not obvious when 10**7 bacteria were introduced by the same mechanism. 3. When 10**5 M. lepraemurium phagocytized into macrophages in vitro were inoculated into the footpad, no remarkable differences were observed in the multiplication of M. lepraemurium in the inoculated footpad compared with the control group. The thickness changes of the footpad injected with 10**5 M. lepraemurium within 20th week after inoculation were not significant. However. in the animals inoculated with 10**7 bacteria, the thickening of injected footpad became obvious since 15th week post-inoculation. 4. It seemed that propagation of M. lepraemurium into the various organs after footpad inoculation cia macroptages in experimental group was faster than that in control group. 5. Belayed hypersensitivity against soluble M. lepraemurium antigen at 17 and 20th week after footpad inoculation of M. lepraemurium was not meaningfully different when that of experimental stoup was compared with that of controls. This reaction seemed to be weakened and disappear from 20th week after footpad inoculation. Therefore, it was tentatively concluded that those enhancement and/or suppression of host defense mechanism due to application of non-viable antigen via macrophageg might not be seen in the experiment in which live M. lepraemurium were introduced into animals via macrophages.
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