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两种在成年大鼠体内内皮细胞中表达受限的CD34变体的免疫靶向与克隆

Immunotargeting and cloning of two CD34 variants exhibiting restricted expression in adult rat endothelia in vivo.

作者信息

Testa Jacqueline E, Chrastina Adrian, Oh Phil, Li Yan, Witkiewicz Halina, Czarny Malgorzata, Buss Tim, Schnitzer Jan E

机构信息

Proteogenomics Research Institute For Systems Medicine, Sidney Kimmel Cancer Center, San Diego, CA 92121, USA.

出版信息

Am J Physiol Lung Cell Mol Physiol. 2009 Aug;297(2):L251-62. doi: 10.1152/ajplung.90565.2008. Epub 2009 May 22.

Abstract

Mapping protein expression of endothelial cells (EC) in vivo is fundamental to understanding cellular function and may yield new tissue-selective targets. We have developed a monoclonal antibody, MAb J120, to a protein expressed primarily in rat lung and heart endothelium. The antigen was identified as CD34, a marker of hematopoietic stem cells and global marker of endothelial cells in human and mouse tissues. PCR-based cloning identified two CD34 variant proteins, full length and truncated, both of which are expressed on luminal endothelial cell plasma membranes (P) isolated from lung. Truncated CD34 predominated in heart P, and neither variant was detected in P from kidney or liver. CD34 in lung was readily accessible to (125)I-J120 inoculated intravenously, and immunohistochemistry showed strong CD34 expression in lung EC. Few microvessels stained in heart and kidney, and no CD34 was detected in vessels of other organs or in lymphatics. We present herein the first complete sequence of a rat CD34 variant and show for the first time that the encoded truncated variant is endogenously expressed on EC in vivo. We also demonstrate that CD34 expression in rat EC, unlike mouse and human, is restricted in its distribution enabling quite specific lung targeting in vivo.

摘要

在体内绘制内皮细胞(EC)的蛋白质表达图谱对于理解细胞功能至关重要,并且可能产生新的组织选择性靶点。我们已经开发出一种针对主要在大鼠肺和心脏内皮中表达的蛋白质的单克隆抗体MAb J120。该抗原被鉴定为CD34,它是造血干细胞的标志物以及人和小鼠组织中内皮细胞的通用标志物。基于PCR的克隆鉴定出两种CD34变体蛋白,全长和截短型,二者均表达于从肺中分离的腔内内皮细胞质膜(P)上。截短型CD34在心脏P中占主导,在肾脏或肝脏的P中均未检测到任何一种变体。静脉注射的(125)I-J120能够轻易地与肺中的CD34结合,免疫组织化学显示肺EC中CD34表达强烈。心脏和肾脏中仅有少量微血管染色,在其他器官的血管或淋巴管中未检测到CD34。我们在此展示大鼠CD34变体的首个完整序列,并首次表明编码的截短变体在体内的EC上内源性表达。我们还证明,与小鼠和人类不同,大鼠EC中的CD34表达在分布上受到限制,从而在体内实现相当特异的肺靶向。

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