Auth M K, Keitzer R A, Scholz M, Blaheta R A, Hottenrott E C, Herrmann G, Encke A, Markus B H
Transplantations-Immunologisches Labor, Klinik für Allgemeinchirurgie, Johann Wolfgang Goethe-Universität, Frankfurt am Main, Germany.
Hepatology. 1993 Sep;18(3):546-55.
Biliary epithelial cells are a primary site of damage in liver allograft rejection and in immunologically mediated diseases such as primary biliary cirrhosis. Human leukocyte antigens and adhesion molecules on the biliary epithelium are associated with T-lymphocytic binding, recognition and destruction. To investigate relevant cellular immunological mechanisms under standard conditions, we have established an in vitro model using human gallbladder epithelial cells. Although not directly affected in these aberrations, gallbladder epithelial cells are excellent objects for immunological investigations. More than 10(8) highly purified cells were isolated and cultured longer than 6 wk in confluent monolayers. Cell growth was routinely established on uncoated plastic culture dishes, and serum-free media could be applied for immunological experiments. Cell characterization was performed by means of specific monoclonal antibodies typical for biliary epithelial cells. Cytokeratins 1 through 8, 18 and 19 and human epithelial cell antibody 125 always showed strong positive staining. Antigen patterns were examined before and after treatment with interferon-gamma by use of immunohistochemical staining methods. Untreated human gallbladder epithelial cells expressed human leukocyte class I antigens but few or no class II antigens. After stimulation with interferon-gamma induction of human leukocyte antigen-DR, -DP and -DQ was detected. In addition, intercellular adhesion molecule 1 was induced on these gallbladder epithelial cells. Therefore an immunological competence similar to that of biliary epithelial cells could be demonstrated. In vitro cell cultures of gallbladder epithelial cells offer a promising tool for subsequent investigations concerning intrahepatic biliary epithelial cells and their interactions with cells of the immune system.
胆管上皮细胞是肝移植排斥反应以及免疫介导疾病（如原发性胆汁性肝硬化）中损伤的主要部位。胆管上皮细胞上的人类白细胞抗原和黏附分子与T淋巴细胞的结合、识别及破坏有关。为了在标准条件下研究相关的细胞免疫机制，我们建立了一种使用人胆囊上皮细胞的体外模型。尽管胆囊上皮细胞在这些异常情况中未直接受到影响，但它们是免疫研究的理想对象。分离出超过10⁸个高度纯化的细胞，并在汇合单层中培养超过6周。细胞生长通常在未包被的塑料培养皿上进行，无血清培养基可用于免疫实验。通过胆管上皮细胞特有的特异性单克隆抗体进行细胞鉴定。细胞角蛋白1至8、18和19以及人上皮细胞抗体125始终显示强阳性染色。使用免疫组织化学染色方法在干扰素-γ处理前后检查抗原模式。未处理的人胆囊上皮细胞表达人类白细胞I类抗原，但很少或不表达II类抗原。用干扰素-γ刺激后，检测到人类白细胞抗原-DR、-DP和-DQ的诱导表达。此外，这些胆囊上皮细胞上诱导表达了细胞间黏附分子1。因此，可以证明其具有与胆管上皮细胞相似的免疫能力。胆囊上皮细胞的体外细胞培养为后续关于肝内胆管上皮细胞及其与免疫系统细胞相互作用的研究提供了一个有前景的工具。
