本章的主题由其两个先前版本的标题“健康和疾病中的人类Hsp60的免疫组化研究：从自身免疫到癌症”所定义，最新版本出版于2018年。此后，在自身免疫和癌症领域取得了可观的进展，其中一些进展与对伴侣系统（CS）的理解进展密切相关。伴侣系统是由分子伴侣、共伴侣、伴侣辅因子和伴侣相互作用物和受体组成的生理系统。分子伴侣是伴侣系统的主要成员，本章着重于其中之一，Hsp60。由于细胞外囊泡（EVs）在伴侣系统的功能和癌变中也成为关键因素，我们已经增加了关于它们的详细部分。本章解释了如何评估组织和EVs中的Hsp60，以应用于诊断、预后和患者监测，并最终用于开发使用它们作为治疗靶点和工具的方法。我们描述了免疫组化技术、免疫荧光和双免疫荧光共聚焦显微镜技术，以及从血浆中收集和分离EVs并评估其中Hsp60和相关微小RNA（miRNAs）的方法。所有这些程序在研究和管理各种类型的癌症、炎症和自身免疫疾病中已被证明是可靠和有用的。© 2023. 作者（们），在Springer Science+Business Media, LLC（属于Springer Nature的一部分）的独家许可下。

The subject matter of this chapter is defined by the title of its two previous editions, "Immunohistochemistry of human Hsp60 in health and disease: From autoimmunity to cancer," the latest of which appeared in 2018. Since then, considerable advances have been made in the fields of autoimmunity and cancer and some of them are closely linked to progress in the understanding of the chaperone system (CS). This is a physiological system composed of molecular chaperones, co-chaperones, chaperone cofactors, and chaperone interactors and receptors. The molecular chaperones are the chief members of the CS, and here we focus on one of them, Hsp60. Since extracellular vesicles (EVs) have also emerged as key factors in the functioning of the CS and in carcinogenesis, we have incorporated a detailed section about them. This chapter explains how to assess Hsp60 in tissues and in EVs for application in diagnosis, prognostication, and patient monitoring and, eventually, for developing methods using them as therapeutic targets and tools. We describe immunohistochemical techniques, immunofluorescence and double immunofluorescence-confocal microscopy, and methods for collecting and isolating EVs from blood plasma and for assessing their contents in Hsp60 and related microRNAs (miRNAs). All these procedures have proven to be reliable and useful in the study and management of various types of cancer and inflammatory and autoimmune conditions.© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.