Barrett A W, Beynon A D, Reid D J
Department of Oral Biology, University of Newcastle-upon-Tyne, UK.
Histochem J. 1994 Feb;26(2):134-41. doi: 10.1007/BF00157962.
An immunoperoxidase technique was used to compare wax-embedded tissue with frozen tissue for quantitative immunohistochemistry of oral mucosal Langerhans cells. Initial experiments using anti-CD1a, -HLADR and -S100 antisera showed that phenotype, fixative, antibody dilution and trypsinisation of the tissue section significantly affected Langerhans cell counts. Only the anti-HLADR antibody detected Langerhans cells in both frozen and wax-embedded sections. Some 38% of S100-positive dendritic cells were situated in the stratum basale, and 41-84% of these contained melanin as determined by double-labelling. Sections from 39 volunteers were then reacted with the anti-CD1a and -HLADR antibodies. The morphology of Langerhans cells was more dendritic in frozen sections, and the mean HLADR-positive Langerhans cells count in frozen sections was significantly higher than that in wax-embedded sections from the same individual. The intra-individual ratio of counts between frozen and wax-embedded sections was variable; hence, the apparent loss of HLADR antigenicity as a result of tissue processing was unpredictable. Counts of CD1a-positive Langerhans cells were consistently higher. We conclude that the use of anti-CD1a antibody on frozen tissue is the optimum method for quantitative studies of oral mucosal Langerhans cells, and that such studies performed on wax-embedded tissue may be unreliable.
采用免疫过氧化物酶技术，对口腔黏膜朗格汉斯细胞进行定量免疫组织化学分析，比较石蜡包埋组织与冷冻组织。最初使用抗CD1a、抗HLADR和抗S100抗血清进行的实验表明，组织切片的表型、固定剂、抗体稀释度和胰蛋白酶处理对朗格汉斯细胞计数有显著影响。只有抗HLADR抗体能在冷冻切片和石蜡包埋切片中均检测到朗格汉斯细胞。约38%的S100阳性树突状细胞位于基底层，通过双重标记确定，其中41 - 84%含有黑色素。随后，对39名志愿者的切片用抗CD1a和抗HLADR抗体进行反应。冷冻切片中朗格汉斯细胞的形态更呈树突状，同一人冷冻切片中HLADR阳性朗格汉斯细胞的平均计数显著高于石蜡包埋切片。冷冻切片与石蜡包埋切片之间的个体内计数比例各不相同；因此，由于组织处理导致的HLADR抗原性明显丧失是不可预测的。CD1a阳性朗格汉斯细胞的计数始终较高。我们得出结论，在冷冻组织上使用抗CD1a抗体是口腔黏膜朗格汉斯细胞定量研究的最佳方法，而在石蜡包埋组织上进行此类研究可能不可靠。
