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Identification of Tertiary Lymphoid Structure-Associated Follicular Helper T Cells in Human Tumors and Tissues

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Tertiary Lymphoid Structures

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1845))

Abstract

Follicular helper T (Tfh) cells are major components of the humoral immune response due to their pivotal role in germinal center formation and antibody affinity maturation following B-cell isotype switching. This CD4+ T-cell subtype is mainly found in the B-cell zone of secondary lymphoid organs as well as in tertiary lymphoid structures (TLS), which are highly organized structures composed of T and B cells, occasionally found at the invasive margin in the tumor microenvironment.

We describe here how to perform immunofluorescence staining of tumor tissue sections and multicolor flow cytometry on tumor cell suspensions to identify and visualize these TLS-associated Tfh cells within the tumor microenvironment of various human cancers. These assays take advantage of combinations of markers and molecules involved in Tfh differentiation and function.

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Abbreviations

Bcl-6:

B-cell lymphoma 6

CXCL13:

C-X-C motif chemokine 13

CXCR5:

C-X-C chemokine receptor type 5

FBS:

Fetal bovine serum

FDC:

Follicular dendritic cell

FFPE:

Formalin-fixed paraffin-embedded

GC:

Germinal center

HCC:

Hepatocellular carcinoma

HS:

Human serum

ICOS:

Inducible costimulator

IF:

Immunofluorescence

LN:

Lymph node

mDC:

Mature dendritic cell

NSCLC:

Non-small cell lung cancer

PBS:

Phosphate-buffered saline

PC:

Plasma cell

PD-1:

Programmed death-1

RT:

Room temperature

SLO:

Secondary lymphoid organ

Tfh:

Follicular helper T cell

TLS:

Tertiary lymphoid structure

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Funding

Hélène Kaplon was supported by a grant from La Ligue contre le Cancer. The work developed by B. Dubois’s group was supported by the Ligue Contre le Cancer Comité de Savoie, Fondation ARC pour la recherche sur le cancer, and INCa (PRT-K16).

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Author notes

  1. Claire Germain

    Present address: Laboratory “Immune Intervention and Biotherapies”, UPMC UMRS CR7—Inserm U1135—CNRS ERL 8255, Centre d’Immunologie et des Maladies Infectieuses (CIMI), Paris, France

Authors and Affiliations

  1. Laboratory “Targeting of the Tumor and Its Immune Microenvironment”, Centre Léon Bérard, Centre de Recherche en Cancérologie de Lyon, University Claude Bernard Lyon 1, Institut National de la Santé et de la Recherche Médicale (INSERM) U1052, CNRS 5286, Lyon, France

    Coline Couillault & Bertrand Dubois

  2. Cordeliers Research Center, Laboratory “Cancer, Immune Control and Escape”, INSERM, UMRS 1138, Paris, France

    Claire Germain & Hélène Kaplon

  3. Cordeliers Research Center, Paris Descartes University, Sorbonne Paris Cité, UMRS 1138, Paris, France

    Claire Germain & Hélène Kaplon

  4. Cordeliers Research Center, Sorbonne University, UMRS 1138, Paris, France

    Claire Germain & Hélène Kaplon

Authors
  1. Coline Couillault
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  2. Claire Germain
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  3. Bertrand Dubois
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  4. Hélène Kaplon
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Corresponding author

Correspondence to Bertrand Dubois .

Editor information

Editors and Affiliations

  1. Laboratory “Cancer, Immune Control and Escape”, UMRS 1138 INSERM, Cordeliers Research Center, Sorbonne University, UMR 1138, Paris Descartes University, Sorbonne Paris City, UMR 1138, Paris, France

    Marie-Caroline Dieu-Nosjean

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Couillault, C., Germain, C., Dubois, B., Kaplon, H. (2018). Identification of Tertiary Lymphoid Structure-Associated Follicular Helper T Cells in Human Tumors and Tissues. 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_12

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  • DOI: https://doi.org/10.1007/978-1-4939-8709-2_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8708-5

  • Online ISBN: 978-1-4939-8709-2

  • eBook Packages: Springer Protocols

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