Nitta M, Kato Y, Strife A, Wachter M, Fried J, Perez A, Jhanwar S, Duigou-Osterndorf R, Chaganti R S, Clarkson B
Blood. 1985 Nov;66(5):1053-61.
Peripheral blood specimens were obtained from 22 patients with Philadelphia chromosome (Ph1) positive chronic myelogenous leukemia (CML) (16 in chronic phase, 2 in an accelerated phase, and 4 in blast crisis). Studies were performed to determine the frequency of the presence of the Ph1 chromosome in cells of lymphoid lineages. Rosetted (E+) lymphocytes (T lymphocytes) from nine patients in chronic phase and one patient in blast crisis were stimulated with T cell growth factor interleukin 2 (IL-2) and/or phytohemagglutinin (PHA). All ten patients had sufficient T lymphocyte metaphases for analysis and of a total of 461 metaphases examined, only one contained the Ph1 chromosome. Nucleated cells of density less than 1.077 g/mL were infected with Epstein-Barr virus (EBV). Following infection, cell lines were established from individual colonies attached to egg albumin-coated Lab-Tek slide chambers (clonal cell lines) or from suspension culture in 96-well tissue culture cluster dishes (nonclonal cell lines). Cell surface and intracellular marker analysis confirmed the B lymphocyte phenotype of all the cell lines examined. B lymphoblastoid cell lines were established from 16 of the 22 patients. All lines from 12 patients were Ph1-negative. From two chronic phase patients, both Ph1-positive and Ph1-negative lines were established. From one patient in an accelerated phase, only Ph1-positive lines were established. From another patient in blast crisis (of myeloblastic phenotype), only Ph1-positive lines were established initially; however, five months later, after the patient had been treated with mitoxantrone, only Ph1-negative lines were derived from this patient. Based on these results, it appears that most B cells and mature T cells in most CML patients are Ph1-negative, but that about 25% of patients have predominantly Ph1-positive B cells or a mixture of Ph1-positive and Ph1-negative B cells that are capable of growing as established cell lines after transformation with EBV.
从22例费城染色体（Ph1）阳性的慢性粒细胞白血病（CML）患者（慢性期16例、加速期2例、急变期4例）获取外周血标本。开展研究以确定淋巴系细胞中Ph1染色体的存在频率。来自9例慢性期患者和1例急变期患者的玫瑰花结形成（E+）淋巴细胞（T淋巴细胞）用T细胞生长因子白细胞介素2（IL-2）和/或植物血凝素（PHA）刺激。所有10例患者都有足够的T淋巴细胞中期分裂相用于分析，在总共检查的461个中期分裂相中，只有1个含有Ph1染色体。密度小于1.077 g/mL的有核细胞用爱泼斯坦-巴尔病毒（EBV）感染。感染后，从附着于卵白蛋白包被的Lab-Tek载玻片培养室的单个集落建立细胞系（克隆细胞系），或从96孔组织培养板中的悬浮培养物建立细胞系（非克隆细胞系）。细胞表面和细胞内标志物分析证实了所有检测细胞系的B淋巴细胞表型。从22例患者中的16例建立了B淋巴母细胞系。12例患者的所有细胞系均为Ph1阴性。从2例慢性期患者中，建立了Ph1阳性和Ph1阴性细胞系。从1例加速期患者中，仅建立了Ph1阳性细胞系。从另1例急变期患者（髓母细胞表型）中，最初仅建立了Ph1阳性细胞系；然而，5个月后，在该患者接受米托蒽醌治疗后，仅从该患者获得了Ph1阴性细胞系。基于这些结果，似乎大多数CML患者的大多数B细胞和成熟T细胞是Ph1阴性的，但约25%的患者主要有Ph1阳性B细胞或Ph1阳性和Ph1阴性B细胞的混合物，这些细胞在用EBV转化后能够作为已建立的细胞系生长。
