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[慢性淋巴细胞白血病患者中白细胞介素2和CpG寡核苷酸DSP30刺激的外周血淋巴细胞的染色体带型分析]

[Chromosome banding analysis of peripheral blood lymphocytes stimulated with IL2 and CpG oligonucleotide DSP30 in patients with chronic lymphocytic leukemia].

作者信息

Stěpanovská K, Vaňková G, Némethová V, Tomášiková L, Smuhařová P, Divíšková E, Vallová V, Kuglík P, Plevová K, Oltová A, Doubek M, Pospíšilová S, Mayer J

机构信息

Interní hematologická a onkologická klinika, Centrum molekulární bio­logie a genové terapie, LF MU a FN Brno.

出版信息

Klin Onkol. 2013;26(4):263-70. doi: 10.14735/amko2013263.

Abstract

BACKGROUND

Chromosomal aberrations play an important role as prognostic factors in chronic lymphocytic leukemia (CLL). These aberrations are mostly detected by fluorescent in situ hybridization (FISH), as chromosomal banding analysis has been scarce due to low proliferative activity of malignant B-lymphocytes in vitro. In 2006, a new method using stimulation with IL-2 and CpG oligonucleotide DSP30 for metaphase generation in CLL was published [1]. The objective of our study was to verify the efficacy of stimulation and to evaluate if the method is suitable for routine diagnostics.

PATIENTS AND METHODS

In total, peripheral blood samples of 369 CLL patients were analyzed in parallel by chromosomal banding analysis and by FISH probes for 13q14, 11q22-23, CEP12 and 17p13.

RESULTS

Out of 369 patients, 307 (83%) were successfully stimulated for metaphase generation. Chromosomal aberrations were detected in 243 (79%) out of 307 patients evaluated by chromosomal banding analysis. Other aberrations that are not included into standard FISH panel were detected in patients karyotypes, e.g. del(6q), del(14q), t(14;18)(q32;q21), t(11;14)(q13;q32) and t(18;22)(q21;q11). One hundred and three (42%) patients showed complex aberrant karyotype not detected by FISH analysis.

CONCLUSION

Stimulation with IL-2 and oligonucleotide DSP30 is an efficient method how to induce proliferation of malignant B-lymphocytes and allows detection of a substantial number of chromosomal aberrations in addition to those detected by standard FISH panel. Using this method in routine diagnostics is helpful particularly in identification of patients with complex aberrant karyotype.

摘要

背景

染色体畸变作为慢性淋巴细胞白血病(CLL)的预后因素发挥着重要作用。这些畸变大多通过荧光原位杂交(FISH)检测,因为由于恶性B淋巴细胞在体外的增殖活性较低,染色体显带分析很少见。2006年,一种使用白细胞介素-2(IL-2)和CpG寡核苷酸DSP30刺激以产生CLL中期相的新方法被发表[1]。我们研究的目的是验证刺激的有效性,并评估该方法是否适用于常规诊断。

患者与方法

总共对369例CLL患者的外周血样本同时进行染色体显带分析以及针对13q14、11q22 - 23、着丝粒12(CEP12)和17p13的FISH探针检测。

结果

在369例患者中,307例(83%)成功被刺激产生中期相。通过染色体显带分析评估的307例患者中有243例(79%)检测到染色体畸变。在患者核型中检测到了标准FISH检测组未包括的其他畸变,例如del(6q)、del(14q)、t(14;18)(q32;q21)、t(11;14)(q13;q32)和t(18;22)(q21;q11)。103例(42%)患者显示出FISH分析未检测到的复杂异常核型。

结论

用IL-2和寡核苷酸DSP30刺激是诱导恶性B淋巴细胞增殖的有效方法,除了标准FISH检测组检测到的那些畸变外,还能检测到大量染色体畸变。在常规诊断中使用该方法特别有助于识别具有复杂异常核型的患者。

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