Abstract
Tertiary lymphoid structures (TLS) are de novo lymphoid formations that are induced within tissues during inflammatory episodes. TLS have been reported at various anatomic sites and in many different contexts like cancer, infections, autoimmunity, graft rejection, and idiopathic diseases. These inducible, ectopic, and transient lymphoid structures exhibit the prototypical architecture found within secondary lymphoid organs (SLO) and have been recently appreciated as a major driver of the local adaptive immune reaction. As TLS emerge within tissues, the isolation in situ and the molecular characterization of these structures are challenging to operate. Laser capture microdissection (LCM) is a powerful tool to isolate selective structural components and cells from frozen or paraffin-embedded tissues. We and other groups previously applied LCM to decipher the molecular network within TLS and uncover their intrinsic connection with the local microenvironment. In this chapter, we describe a detailed LCM method for selecting and isolating TLS in situ to perform comprehensive downstream molecular analyses.
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Acknowledgments
We thank all present and past members of the Laboratory “Cancer, Immune control and Escape” (UMRS1138 INSERM, Cordeliers Research Center), pathologists, and clinicians who have participated in these studies, for their help and valuable discussions. We are grateful to lung cancer patients involved in this research performed in our laboratory.
Funding: This work was supported by the Institut National de la Santé et de la Recherche Médicale (INSERM), Pierre and Marie Curie University, Paris-Descartes University, the Labex Immuno-Oncology (LAXE62_9UMRS872 Fridman), CARPEM (Cancer Research for PErsonalized Medicine), Fondation ARC pour la Recherche sur le Cancer. Jeremy Goc was supported by a grant from the Fondation ARC pour la Recherche sur le Cancer.
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Gutierrez-Chavez, C., Knockaert, S., Dieu-Nosjean, MC., Goc, J. (2018). Development of Methods for Selective Gene Expression Profiling in Tertiary Lymphoid Structure Using Laser Capture Microdissection. In: Dieu-Nosjean, MC. (eds) Tertiary Lymphoid Structures. Methods in Molecular Biology, vol 1845. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8709-2_8
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DOI: https://doi.org/10.1007/978-1-4939-8709-2_8
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