Nap M, Vitali A, Nustad K, Bast R C, O'Brien T J, Nilsson O, Seguin P, Suresh M R, Børmer O P, Saga T, de Bruijn H W, Nozawa S, Kreutz F T, Jette D, Sakahara H, Gadnell M, Endo K, Barlow E H, Warren D, Paus E, Hammarström S, Kenemans P, Hilgers J
Institute for Public Health, Department of Pathology, Leeuwarden, The Netherlands.
Tumour Biol. 1996;17(6):325-31.
We evaluated the immunohistological (IH) characteristics of 22 different antibodies that were submitted for study in the frame of the TD-1 ISOBM Workshop on monoclonal antibodies against CA125. Information on relative affinities and epitope similarities was obtained from a parallel immunochemical study. Antibodies were tested at concentrations of 10 and 1 micrograms/ml on frozen and paraffin sections. Paraffin sections were stained according to the streptavidin-biotin complex protocol, and frozen sections according to a two-step immunoperoxidase technique. Aminoethylcarbazole served as the chromogen. The tissues were from normal proliferative endometrium (formalin-fixed paraffin-embedded) material and clear-cell adenocarcinoma of the ovary (formalin-fixed paraffin-embedded and frozen material). Sections were scored for staining in epithelial cells, basal, apical and diffuse cytoplasmic and in stromal components. Intensity was graded as 1, 2 or 3 for epithelial cells and as -1, -2 or -3 for stroma. The cumulative scores for each antibody expressed the discriminative properties of specific epithelial staining against background. M11 and M11-like antibodies, as well as OC125 and OC125-like antibodies, in general showed good staining results. Although there was a trend for high-affinity antibodies to show higher scores, there was no clear relationship between affinity and staining result. For nine antibodies (ZR45, MA602-1, K102, K94, K90, OV185, K97, K96, OV198), the reactions in paraffin and frozen sections were of similar intensity. Most of these were of low affinity with one exception: antibody ZR45, a rat monoclonal antibody (MAb) which had a high relative affinity. For eight antibodies (M11, K101, MA602-6, ZS33, B27.1, B43.13, K93, OC125), a loss of specific staining was observed in frozen sections. All but two of these antibodies (MA602-6 and OC125) were of high relative affinity. With four antibodies (K91, ZR38, K95, K100), the reverse situation was observed. One (K100) was of low affinity, two (K95 and K91) of high affinity and the fourth (ZR38) was a rat MAb of high affinity. Mainly due to the increased cytoplasmic staining in carcinoma, the reactivity in paraffin sections was less extensive in normal endometrium compared to ovarian carcinoma for the majority of antibodies, irrespective of their affinity or epitope group. The IH characterization of these antibodies may be of help in selecting antibodies with specific properties for further comparative studies. The reactivity of normal endometrium with all useful antibodies makes it a good candidate for standard external IH tissue control.
我们评估了22种不同抗体的免疫组织学（IH）特征，这些抗体是在TD-1 ISOBM抗CA125单克隆抗体研讨会上提交用于研究的。相对亲和力和表位相似性的信息来自一项平行免疫化学研究。抗体分别以10微克/毫升和1微克/毫升的浓度在冰冻切片和石蜡切片上进行检测。石蜡切片按照链霉亲和素-生物素复合物方案染色，冰冻切片按照两步免疫过氧化物酶技术染色。氨基乙基咔唑用作显色剂。组织来自正常增殖期子宫内膜（福尔马林固定石蜡包埋）材料和卵巢透明细胞腺癌（福尔马林固定石蜡包埋及冰冻材料）。对上皮细胞、基底、顶端和弥漫性细胞质以及基质成分的染色进行评分。上皮细胞强度分为1、2或3级，基质分为-1、-2或-3级。每种抗体的累积分数表示特异性上皮染色相对于背景的鉴别特性。M11和M11样抗体以及OC125和OC125样抗体总体上显示出良好的染色结果。尽管高亲和力抗体有显示更高分数的趋势，但亲和力与染色结果之间没有明确的关系。对于9种抗体（ZR45、MA602-1、K102、K94、K90、OV185、K97、K96、OV198），在石蜡切片和冰冻切片中的反应强度相似。其中大多数亲和力较低，只有一个例外：大鼠单克隆抗体（MAb）ZR45具有较高的相对亲和力。对于8种抗体（M11、K101、MA60-6、ZS33、B27.1、B43.13、K93、OC125），在冰冻切片中观察到特异性染色丧失。除两种抗体（MA602-6和OC125）外，其余抗体均具有较高的相对亲和力。对于4种抗体（K91、ZR38、K95、K100），观察到相反的情况。一种（K100）亲和力低，两种（K95和K91）亲和力高，第四种（ZR38）是高亲和力的大鼠MAb。主要由于癌组织中细胞质染色增加，与卵巢癌相比，大多数抗体在正常子宫内膜石蜡切片中的反应性范围较小，无论其亲和力或表位组如何。这些抗体的IH特征可能有助于选择具有特定特性的抗体进行进一步的比较研究。正常子宫内膜与所有有用抗体的反应性使其成为标准外部IH组织对照的良好候选者。
