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[“经典”巨噬细胞标志物CD68在原代人成纤维细胞中强烈表达]

[The "classical" macrophage marker CD68 is strongly expressed in primary human fibroblasts].

作者信息

Kunz-Schughart L A, Weber A, Rehli M, Gottfried E, Brockhoff G, Krause S W, Andreesen R, Kreutz M

机构信息

Institute of Pathology, University of Regensburg, Germany.

出版信息

Verh Dtsch Ges Pathol. 2003;87:215-23.

Abstract

AIM

Monoclonal antibodies against the human homologue of mouse macrosialin, CD68, are generally commercialized as markers for human monocytes and macrophages. Indeed, CD68 is considered as a selective marker for human myeloid cells, although several previous immunohistochemical studies indicate that some antibody clones also react with other hematopoietic and non-hematopoietic cell types. The aim of our study was to verify these observations and to evaluate the reliability of CD68 as a macrophage marker.

METHODS

We investigated protein and RNA expression of CD68 in various fibroblast types and carcinoma cell lines as compared to monocytes and macrophages using immunohistochemistry, flow cytometry, and specific RT-PCR. Different monoclonal antibody clones against CD68 were applied including KP-1 and EBM11.

RESULTS

As expected, the intensity of immunohistochemical and flow cytometric CD68 staining was dependent on both the antibody clone and the fixation procedure. However, fibroblasts isolated from normal skin, normal breast, breast tumor tissue, and osteoarthritis synovia clearly expressed CD68 protein at levels comparable to macrophages. The specificity of CD68 expression in fibroblasts was verified by RT-PCR which also showed some tumor cell types to express CD68 mRNA.

CONCLUSION

Our findings clearly demonstrate that the expression of CD68 is not restricted to the macrophage lineage. This is highly relevant for experimental and diagnostic purposes, since anti CD68 antibodies cannot be accepted without reservations for the discrimination of myeloid cells and fibroblasts even in paraffin sections after formalin fixation.

摘要

目的

针对小鼠巨唾液酸蛋白的人类同源物CD68的单克隆抗体通常作为人类单核细胞和巨噬细胞的标志物商业化。实际上,CD68被认为是人类髓样细胞的选择性标志物,尽管先前的一些免疫组织化学研究表明,一些抗体克隆也与其他造血和非造血细胞类型发生反应。我们研究的目的是验证这些观察结果,并评估CD68作为巨噬细胞标志物的可靠性。

方法

我们使用免疫组织化学、流式细胞术和特异性逆转录聚合酶链反应(RT-PCR),将单核细胞和巨噬细胞与各种成纤维细胞类型和癌细胞系中的CD68蛋白和RNA表达进行了比较。应用了不同的针对CD68的单克隆抗体克隆,包括KP-1和EBM11。

结果

正如预期的那样,免疫组织化学和流式细胞术CD68染色的强度取决于抗体克隆和固定程序。然而,从正常皮肤、正常乳腺、乳腺肿瘤组织和骨关节炎滑膜中分离出的成纤维细胞明显表达CD68蛋白,其水平与巨噬细胞相当。通过RT-PCR验证了成纤维细胞中CD68表达的特异性,该方法还显示一些肿瘤细胞类型表达CD68 mRNA。

结论

我们的研究结果清楚地表明,CD68的表达不限于巨噬细胞谱系。这对于实验和诊断目的高度相关,因为即使在福尔马林固定后的石蜡切片中,抗CD68抗体也不能毫无保留地用于区分髓样细胞和成纤维细胞。

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