Evaluation of somatostatin receptor subtype expression in human neuroendocrine tumors using two sets of new monoclonal antibodies

Regulatory Peptides Pub Date : 2013-11-10 DOI:10.1016/j.regpep.2013.10.007

Chiara Lambertini , Patrizia Barzaghi-Rinaudo , Lisa D'Amato , Stefan Schulz , Paolo Nuciforo , Herbert A. Schmid

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引用次数: 22

Abstract

Introduction

The expression and reliable detection of somatostatin receptor subtypes (SSTR1–5) is a prerequisite for the successful use of somatostatin analogs in neuroendocrine tumors (NETs). Two sets of monoclonal antibodies (mAbs) against human SSTR1, 2A, 3 and 5 have recently been developed by two independent laboratories using rabbit and mouse hybridomas. Our aim was to evaluate the usefulness of both sets of mAbs for detection of SSTRs in NET samples as they are routinely collected in clinical practice.

Methods

Mouse and rabbit mAbs were characterized in SSTR1, 2A, 3 and 5-transfected HEK293 cells and human archival samples of pancreatic tissue and NET. Comparative analysis of mAbs was also conducted by immunostaining of a tissue microarray composed of 75 cores of NET.

Results

Immunohistochemical analysis of HEK293 cells showed that both rabbit and mouse mAbs specifically detect their cognate receptor subtype, with mild cytoplasmic cross-reactivity observed for rabbit mAbs. Both sets of mAbs labeled normal pancreatic islets and showed similar patterns of immunoreactivity in NET controls. Direct comparison of mAb sets using a NET tissue microarray revealed strong correlation between rabbit and mouse mAbs against SSTR1 and 5, and moderate correlation for SSTR3. The rabbit mAb against SSTR2A showed higher affinity for its cognate receptor than the corresponding mouse mAb, resulting in a more reliable detection of this SSTR.

Conclusions

mAbs from both sets are reliable tools for the detection of SSTR1, 3 and 5, whereas the rabbit mAb against SSTR2A is recommended for use in routine clinical testing due to its superior binding affinity.

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利用两组新的单克隆抗体评价生长抑素受体亚型在人神经内分泌肿瘤中的表达

生长抑素受体亚型(SSTR1-5)的表达和可靠检测是在神经内分泌肿瘤(NETs)中成功使用生长抑素类似物的先决条件。最近，两个独立的实验室利用兔和小鼠杂杂瘤开发了两组针对人SSTR1、2A、3和5的单克隆抗体(mab)。我们的目的是评估两组单克隆抗体在NET样本中检测sstr的有效性，因为它们在临床实践中是常规收集的。方法在SSTR1、2A、3和5转染的HEK293细胞以及人胰腺组织和NET档案样本中检测小鼠和兔单抗。通过对由75个NET核组成的组织微阵列进行免疫染色，对单克隆抗体进行比较分析。结果对HEK293细胞的免疫组化分析显示，兔单抗和小鼠单抗均能特异性检测其同源受体亚型，兔单抗具有轻微的细胞质交叉反应性。两组单克隆抗体标记正常胰岛，并在NET对照中显示相似的免疫反应模式。使用NET组织微阵列直接比较单克隆抗体组发现兔和小鼠单克隆抗体对SSTR1和5具有很强的相关性，对SSTR3具有中等相关性。兔抗SSTR2A单抗对其同源受体的亲和力高于相应的小鼠单抗，从而更可靠地检测该SSTR。结论两组单抗都是检测SSTR1、3和5的可靠工具，而针对SSTR2A的兔单抗由于其优越的结合亲和力，推荐用于常规临床检测。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Regulatory Peptides 医学-内分泌学与代谢

自引率

0.00%

发文量

审稿时长

2 months

期刊介绍： Regulatory Peptides provides a medium for the rapid publication of interdisciplinary studies on the physiology and pathology of peptides of the gut, endocrine and nervous systems which regulate cell or tissue function. Articles emphasizing these objectives may be based on either fundamental or clinical observations obtained through the disciplines of morphology, cytochemistry, biochemistry, physiology, pathology, pharmacology or psychology.