Reassessment of CXCR4 chemokine receptor expression in human normal and neoplastic tissues using the novel rabbit monoclonal antibody UMB-2.
Reassessment of CXCR4 chemokine receptor expression in human normal and neoplastic tissues using the novel rabbit monoclonal antibody UMB-2.
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DOI：
10.1371/journal.pone.0004069
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发表时间：
2008
期刊：
影响因子：
3.7
通讯作者：
Schulz, Stefan
中科院分区：
文献类型：
--
作者：
Fischer, Thomas;Nagel, Falko;Jacobs, Stefan;Stumm, Ralf;Schulz, Stefan
CXCR4趋化因子受体调节癌细胞向特定转移部位的迁移和归巢。确定CXCR4受体状态将为疾病预后和可能的治疗干预提供预测性信息。然而，先前使用特性不明确的小鼠单克隆或兔多克隆抗体定位CXCR4的尝试产生了主要的核染色和偶尔的细胞质染色，但没有发现真正的细胞表面受体。在本研究中，我们利用cxcr4缺陷小鼠的转染细胞和组织广泛表征了新型兔单克隆抗cxcr4抗体（克隆mb -2）。转染cxcr4的细胞表面染色证实了mb -2的特异性；激动剂暴露后CXCR4免疫染色易位；cxcr4表达细胞匀浆的Western blot检测到M r 38,000-43,000的宽带迁移；在CXCR4+/+小鼠组织中选择性检测受体，而在CXCR4−/−小鼠组织中没有选择性检测受体；用其免疫肽预吸附mb -2来消除组织免疫染色。在福尔马林固定的石蜡包埋的人肿瘤组织中，mb -2对肿瘤细胞亚群进行了高效的质膜染色，这些肿瘤细胞通常分布在整个肿瘤中。小鼠单克隆抗体12G5和其他常用的市售抗体的比较分析显示，在其他相同的条件下，这些抗体都不能检测到CXCR4。因此，兔单克隆抗体mb -2可能在常规组织病理学检查中评估多种人类肿瘤中CXCR4受体状态方面具有重要价值。
The CXCR4 chemokine receptor regulates migration and homing of cancer cells to specific metastatic sites. Determination of the CXCR4 receptor status will provide predictive information for disease prognosis and possible therapeutic intervention. However, previous attempts to localize CXCR4 using poorly characterized mouse monoclonal or rabbit polyclonal antibodies have produced predominant nuclear and occasional cytoplasmic staining but did not result in the identification of bona fide cell surface receptors. In the present study, we extensively characterized the novel rabbit monoclonal anti-CXCR4 antibody (clone UMB-2) using transfected cells and tissues from CXCR4-deficient mice. Specificity of UMB-2 was demonstrated by cell surface staining of CXCR4-transfected cells; translocation of CXCR4 immunostaining after agonist exposure; detection of a broad band migrating at M r 38,000–43,000 in Western blots of homogenates from CXCR4-expressing cells; selective detection of the receptor in tissues from CXCR4+/+ but not from CXCR4−/− mice; and abolition of tissue immunostaining by preadsorption of UMB-2 with its immunizing peptide. In formalin-fixed, paraffin-embedded human tumor tissues, UMB-2 yielded highly effective plasma membrane staining of a subpopulation of tumor cells, which were often heterogeneously distributed throughout the tumor. A comparative analysis of the mouse monoclonal antibody 12G5 and other frequently used commercially available antibodies revealed that none of these was able to detect CXCR4 under otherwise identical conditions. Thus, the rabbit monoclonal antibody UMB-2 may prove of great value in the assessment of the CXCR4 receptor status in a variety of human tumors during routine histopathological examination.
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影响因子：
11.5
作者：
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通讯作者：
Lai, MaoDe
影响因子：
11.5
作者：
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通讯作者：
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影响因子：
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作者：
Schmid, BC;Rudas, M;Zeillinger, R
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作者：
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DOI：
10.1111/j.1525-1438.2007.00841.x
发表时间：
2007-05-01
影响因子：
4.8
作者：
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通讯作者：
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