Hessel H, Mittermüller J, Zitzelsberger H, Weier H U, Bauchinger M
GSF-National Research Center for Environment and Health, Institute of Radiobiology.
Histochem Cell Biol. 1996 Nov;106(5):481-5. doi: 10.1007/BF02473310.
Langerhans cells (LC) of the skin represent bone marrow-derived dendritic antigen-presenting cells and are therefore important in pathophysiological processes such as rejection, graft-versus-host disease, and graft-versus-leukemia-reaction after bone marrow transplantation (BMT). For understanding of these diseases, the evaluation of the chimeric status of LC following BMT is of great interest. To analyze the sex chromosome constitution of LC in the skin, we established a modified and refined technique of combined immunophenotyping and fluorescence in situ hybridization (FISH) and investigated frozen sections of skin biopsies from nine patients after allogeneic sex-mismatched BMT and of two healthy donors for control. LC were specifically labeled using a fluorescent CD1 a antibody and hybridized simultaneously with X and Y chromosome-specific DNA probes. The results of this practical application on nine leukemia patients show the appearance of donor-type LC and the persistence of host-type LC at various times (36 up to 1395 days) after sex-mismatched BMT. Complete chimerism of LC could not be detected in any case. The frequency of recipient-specific LC ranged from 7% to 92% and showed no correlation with time postgrafting. We conclude from our results of 1461 analyzed LC that combined immunophenotyping and interphase cytogenetic analysis by FISH is the method of choice for the assessment of chimerism in a particular cell type after sex-mismatched BMT. Its practical application on other tissues affected by BMT-related pathophysiological processes reveals further knowledge of the time-dependent course of chimeric patterns after BMT.
皮肤中的朗格汉斯细胞（LC）是源自骨髓的树突状抗原呈递细胞，因此在诸如骨髓移植（BMT）后的排斥反应、移植物抗宿主病和移植物抗白血病反应等病理生理过程中具有重要作用。为了理解这些疾病，评估BMT后LC的嵌合状态备受关注。为了分析皮肤中LC的性染色体组成，我们建立了一种改良和优化的联合免疫表型分析和荧光原位杂交（FISH）技术，并对9例接受异基因性别不匹配BMT的患者以及2例健康供体的皮肤活检冷冻切片进行了研究作为对照。使用荧光CD1a抗体特异性标记LC，并同时与X和Y染色体特异性DNA探针杂交。对9例白血病患者的实际应用结果显示，在性别不匹配BMT后的不同时间（36天至1395天）出现了供体型LC和宿主型LC的持续存在。在任何情况下均未检测到LC的完全嵌合。受体特异性LC的频率范围为7%至92%，且与移植后时间无关。根据我们对1461个分析的LC的结果，我们得出结论，联合免疫表型分析和FISH的间期细胞遗传学分析是评估性别不匹配BMT后特定细胞类型嵌合状态的首选方法。其在受BMT相关病理生理过程影响的其他组织上的实际应用揭示了BMT后嵌合模式随时间变化过程的更多知识。
