Proto-Siqueira Rodrigo, Figueiredo-Pontes Lorena L, Panepucci Rodrigo A, Garcia Aglair B, Rizzatti Edgar G, Nascimento Fabio M, Ishikawa Hellen C F, Larson Roy E, Falcão Roberto P, Simpson Andrew J, Gout Ivan, Filonenko Valery, Rego Eduardo M, Zago Marco A
Hematology Division and Center for Cell Based Therapy, Medical School of Ribeirão Preto, University of São Paulo, Brazil.
Leuk Res. 2006 Nov;30(11):1333-9. doi: 10.1016/j.leukres.2006.02.031. Epub 2006 Apr 18.
The preferentially expressed antigen in melanoma (PRAME) gene is aberrantly expressed in chronic lymphoproliferative disorders (CLD). We produced and characterized an anti-PRAME monoclonal antibody (MoAb), which was then applied in a quantitative flow cytometric (QFC) method to evaluate PRAME expression in leukemic cells from the peripheral blood (PB) of 47 patients with chronic lymphocytic leukemia and seven with mantle cell lymphoma as well as in the PB mononuclear cells (PBMCs) and B lymphocytes from 15 healthy subjects. Approximately 90% of CLD, but none of the normal samples, presented more than 20% of PRAME+ lymphocytes. Moreover, the intensity of PRAME expression was significantly higher in CLD cells compared to normal B lymphocytes and PBMCs. By immunofluorescence microscopy and by permeabilized flow cytometry we demonstrated that PRAME is a membrane antigen and a cytoplasmic protein aberrantly expressed in malignant CLD. Our results suggest that the analysis of PRAME protein may contribute for the distinction between normal and leukemic cells in CLD, and that PRAME may be a potential target for therapy.
黑色素瘤优先表达抗原(PRAME)基因在慢性淋巴细胞增殖性疾病(CLD)中异常表达。我们制备并鉴定了一种抗PRAME单克隆抗体(MoAb),然后将其应用于定量流式细胞术(QFC)方法,以评估47例慢性淋巴细胞白血病患者和7例套细胞淋巴瘤患者外周血(PB)白血病细胞以及15名健康受试者PB单个核细胞(PBMC)和B淋巴细胞中PRAME的表达。约90%的CLD患者样本(但正常样本均无)呈现超过20%的PRAME+淋巴细胞。此外,与正常B淋巴细胞和PBMC相比,CLD细胞中PRAME的表达强度显著更高。通过免疫荧光显微镜和通透流式细胞术,我们证明PRAME是一种膜抗原和细胞质蛋白,在恶性CLD中异常表达。我们的结果表明,PRAME蛋白分析可能有助于区分CLD中的正常细胞和白血病细胞,并且PRAME可能是一个潜在的治疗靶点。