Immunological mechanism on the generation of alcoholic liver diseases

为探讨酒精性肝病发生的免疫学机制，从乙醛诱导抗原性的角度，检测肝脏巨噬细胞（Kupffer cell，巨噬细胞）的作用，并检测酒精作用后肝细胞表面的1类MHC，将胶原酶灌流分离的Kupffer细胞和肝细胞按比重分离后，与乙醛孵育。将这些细胞与针对乙酰氨基酚修饰的同基因细胞（CTL）的细胞毒性T淋巴细胞混合。由于Kupffer细胞表面存在MHC Ⅰ类分子，故Kupffer细胞仅被裂解。这一结果表明，类MHC在免疫反应中的重要性和缺乏的类MHC在正常肝细胞。然后，从类MHC的诱导的角度来检查乙醇给药。饮酒后12 h用细胞毒T淋巴细胞法检测MHC Ⅰ类分子。MHC Ⅰ类分子水平升高， 关于我们 从14天的时间和乙醇的消耗量来看，乙醇组小鼠肝细胞表面未检测到乙醛抗原表位。然后对两组小鼠进行等热量的含或不含乙醇的配对喂养。小鼠喂养至28天，通过胶原酶灌注法分离肝细胞。来自无乙醇喂养的小鼠的肝细胞在其表面上没有显示1类MHC或乙醛表位。乙醇组小鼠肝细胞MHC Ⅰ类分子水平升高，但未检测到CTL所显示的乙醛加合物表位。接下来，将氰胺通过腹腔注射给小鼠，以提高乙醛的血清水平。对用或不用氰胺处理的小鼠进行配对喂养，并与上述相同地检查肝细胞。经氨腈处理的小鼠肝细胞仅显示1类MHC水平升高，因此，长期摄入乙醇和/或预先进行病毒感染等处理可能是肝细胞上乙醛蛋白加合物形成的必要条件。这种推测将在我们的单位用同样的程序进行检查。少

To investigate immunological mechanism on the generation of alcoholic liver diseases, the role of Kupffer cell (macrophage in the liver) was checked from the point of the induction of antigenicity of acetaldehyde, and class 1 MHC on the surface of hepatocyte was checked after administration of alcohol.Kupffer cell and hepatocyte those were isolated by collagenase perfusion and separated according to the specific gravity were incubated with acetaldehyde. Those cells were mixed with cytotoxic T lymphocyte against acetaldehyde-modified syngeneic cells (CTL). Only Kupffer cells were lysed with the reason of the existence of Class 1 MHC on its surface. This result showed the importance of class 1 MHC in immunological reaction and the lack of class 1 MHC on normal hepatocyte.Then, ethanol administration was checked from the point of induction of class 1 MHC. Class 1 MHC was detected by cytotoxic T lymphocyte method shortly 12 hours after alcohol drinking. The level of class 1 MHC elevated ac … More cording to the time up to 14 days and the comsumed volume of ethanol, but acetaldehyde epitope wasn't detected on the surface of hepatocyte from mice that were fed with ethanol. Then pair feeding with or without ethanol was done to two groups of mice isocalolically. Mice were fed up to 28 days, and hepatocytes were isolated by the method of collagenase infusion. Hepatocytes from mice fed without ethanol showed no class 1 MHC or acetaldehyde epitope on its surface. Hepatocytes from mice fed with ethanol revealed the elevated level of class 1 MHC, but no acetaldehyde adduct epitope which would be showed by CTL. Next, the cyanamide was given to mice intraperitoneally to elevate the serum level of acetaldehyde. Pair feeding was done to mice treated with or without cyanamide, and hepatocytes were checked just as same as above. Hepatocytes from mice treated with cyanamide showed only the elevated level of class 1 MHC.So, long term feeding with ethanol and/or previous treatment with such as viral infection might be necessary to build acetaldehyde protein adduct on hepatocytes. Such speculation will be checked with same procedure in our unit. Less