Phospho-specific flow cytometry identifies aberrant signaling in indolent B-cell lymphoma.
Phospho-specific flow cytometry identifies aberrant signaling in indolent B-cell lymphoma.
复制标题
DOI：
10.1186/1471-2407-12-478
复制
发表时间：
2012-10-16
期刊：
影响因子：
3.8
通讯作者：
Kolstad A
中科院分区：
文献类型：
--
作者：
Blix ES;Irish JM;Husebekk A;Delabie J;Forfang L;Tierens AM;Myklebust JH;Kolstad A
恶性细胞信号通路的知识可以提供预后和诊断信息，除了确定潜在的分子靶点治疗。B细胞受体（BCR）和辅助受体CD 40信号传导是正常B细胞所必需的，越来越多的证据表明通过BCR和CD 40的信号传导在B细胞淋巴瘤的发病机制中起重要作用。本研究的目的是研究小细胞淋巴细胞性淋巴瘤/慢性淋巴细胞性白血病（SLL/CLL）和边缘区淋巴瘤（MZL）患者活检单细胞悬液中淋巴瘤B细胞和浸润性T细胞的基础和诱导信号。通过磷酸特异性流式细胞术检查来自未治疗的SLL/CLL和MZL患者的样品的基础和活化诱导的信号传导。靶向B和T细胞的一组9种刺激条件，包括B细胞受体（BCR）、CD 40配体和白细胞介素的交联以及12种匹配的磷蛋白读数，用于研究信号传导。与健康供体B细胞相比，来自SLL/CLL患者的恶性B细胞具有更高的磷酸化（p）-SFKs、p-PLCγ、p-ERK、p-p38、p-p65（NF-κB）、p-STAT 5和p-STAT 6的基础水平。相反，抗BCR诱导的信号在SLL/CLL和MZL B细胞中高度受损，如通过低p-SFK、p-SYK和p-PLCγ水平所确定的。抗BCR诱导的p-PLCγ受损与IgM和CD 79 b的表面表达降低相关。同样，淋巴瘤B细胞中CD 40 L诱导的p-ERK和p-p38也显著降低，而p-p65（NF-κB）与正常B细胞相同。相比之下，IL-2、IL-7和IL-15在肿瘤浸润性T细胞中诱导的p-STAT 5与正常T细胞没有不同。在SLL/CLL和MZL患者的恶性B细胞中，BCR信号传导和CD 40 L诱导的p-p38被抑制。用于检测不同细胞群中信号蛋白的基础以及活化诱导的磷酸化的单细胞磷酸特异性流式细胞术可用于鉴定异常信号通路。
Knowledge about signaling pathways in malignant cells may provide prognostic and diagnostic information in addition to identify potential molecular targets for therapy. B-cell receptor (BCR) and co-receptor CD40 signaling is essential for normal B cells, and there is increasing evidence that signaling via BCR and CD40 plays an important role in the pathogenesis of B-cell lymphoma. The aim of this study was to investigate basal and induced signaling in lymphoma B cells and infiltrating T cells in single-cell suspensions of biopsies from small cell lymphocytic lymphoma/chronic lymphocytic leukemia (SLL/CLL) and marginal zone lymphoma (MZL) patients. Samples from untreated SLL/CLL and MZL patients were examined for basal and activation induced signaling by phospho-specific flow cytometry. A panel of 9 stimulation conditions targeting B and T cells, including crosslinking of the B cell receptor (BCR), CD40 ligand and interleukins in combination with 12 matching phospho-protein readouts was used to study signaling. Malignant B cells from SLL/CLL patients had higher basal levels of phosphorylated (p)-SFKs, p-PLCγ, p-ERK, p-p38, p-p65 (NF-κB), p-STAT5 and p-STAT6, compared to healthy donor B cells. In contrast, anti-BCR induced signaling was highly impaired in SLL/CLL and MZL B cells as determined by low p-SFK, p-SYK and p-PLCγ levels. Impaired anti-BCR-induced p-PLCγ was associated with reduced surface expression of IgM and CD79b. Similarly, CD40L-induced p-ERK and p-p38 were also significantly reduced in lymphoma B cells, whereas p-p65 (NF-κB) was equal to that of normal B cells. In contrast, IL-2, IL-7 and IL-15 induced p-STAT5 in tumor-infiltrating T cells were not different from normal T cells. BCR signaling and CD40L-induced p-p38 was suppressed in malignant B cells from SLL/CLL and MZL patients. Single-cell phospho-specific flow cytometry for detection of basal as well as activation-induced phosphorylation of signaling proteins in distinct cell populations can be used to identify aberrant signaling pathways.
登录
查看更多内容
影响因子：
20.3
作者：
Hewamana, Saman;Alghazal, Suhair;Pepper, Chris
通讯作者：
Pepper, Chris
影响因子：
6.5
作者：
Kater, AP;Evers, LM;Eldering, E
通讯作者：
Eldering, E
影响因子：
4.4
作者：
Irish, Jonathan M.;Czerwinski, Debra K.;Levy, Ronald
通讯作者：
Levy, Ronald
影响因子：
78.5
作者：
Irish, JM;Kotecha, N;Nolan, GP
通讯作者：
Nolan, GP
影响因子：
11.4
作者：
Decker, T;Schneller, F;Peschel, C
通讯作者：
Peschel, C