Thiele J, Hoefer M, Kvasnicka H M, Bertsch H P, Zankovich R, Fischer R
Institute of Pathology University of Cologne, Germany.
Hematol Pathol. 1993;7(4):239-49.
An immunohistochemical and morphometric analysis was performed on trephine biopsy specimens in 60 patients with chronic myeloid leukemia (CML) to quantify erythropoiesis and its proliferation capacity and to assess the stainable marrow iron (hemosiderin). For this purpose, an elaborate double-immunostaining technique was applied. This included a monoclonal antibody (PC10) that is directed against proliferating cell nuclear antigen (PCNA), followed by an antibody against glycophorin C (Ret40f), to identify all nucleated erythroid precursor cells. Additionally, morphometric data were derived from immunostaining of megakaryocytes (CD61) and macrophages (PG-M1), including its hemosiderin-laden subpopulation. Finally the determination of argyrophilic (reticulin) fiber density was carried out. In comparison with a control group (15 patients) without any hematologic disorder, in CML patients morphometric evaluation showed a significant reduction in the number of erythroblasts and normoblasts. This feature was associated with a PCNA-labeling index within the normal range and a decreased stainable marrow iron (number of hemosiderin-storaging macrophages). Several parameters were established to exert a predictive value on survival. A worsening of prognosis was associated with a decrease in the number of erythroid precursors (< 460/mm2), a low hemoglobin level (< 10 g/dl), a high megakaryocyte count (> 50 cells/mm2), an increased density of reticulin fibers (> 30 i x 10(2)/mm2) and splenomegaly (> 15 cm below costal margin). Our findings are in keeping with results obtained from in vitro studies of cell proliferation in CML, which is not significantly altered in comparison with the normal bone marrow. Finally, the present data, although derived from a small group of patients, emphasize the impact of histologic variables to be included in one of the major clinical trials on prognosis in CML.
对60例慢性髓性白血病（CML）患者的环钻活检标本进行了免疫组织化学和形态计量分析，以量化红细胞生成及其增殖能力，并评估可染色的骨髓铁（含铁血黄素）。为此，应用了一种精细的双重免疫染色技术。这包括一种针对增殖细胞核抗原（PCNA）的单克隆抗体（PC10），随后是一种针对血型糖蛋白C（Ret40f）的抗体，以识别所有有核红细胞前体细胞。此外，形态计量数据来自巨核细胞（CD61）和巨噬细胞（PG-M1）的免疫染色，包括其含铁血黄素的亚群。最后进行了嗜银（网状）纤维密度的测定。与无任何血液系统疾病的对照组（15例患者）相比，CML患者的形态计量评估显示早幼红细胞和正成红细胞数量显著减少。这一特征与正常范围内的PCNA标记指数以及可染色骨髓铁（含铁血黄素储存巨噬细胞数量）减少有关。确定了几个对生存具有预测价值的参数。预后恶化与红细胞前体细胞数量减少（<460/mm2）、血红蛋白水平低（<10 g/dl）、巨核细胞计数高（>50个细胞/mm2）、网状纤维密度增加（>30 ix 10(2)/mm2）和脾肿大（肋缘下>15 cm）有关。我们的研究结果与CML细胞增殖的体外研究结果一致，与正常骨髓相比，其增殖没有显著改变。最后，目前的数据虽然来自一小群患者，但强调了组织学变量在CML预后的一项主要临床试验中应被纳入的影响。
