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Rabant M, Calvani J, Terada M, et al. Immunofluorescence multiparamétrique in situ : vers l’amélioration du phénotype de l’infiltrat cellulaire au cours du rejet d’allogreffe rénale. In situ multiplex immunofluorescence analysis of the inflammatory burden in kidney allograft rejection: A new tool to characterize the alloimmune response. Nephrol Ther. 2019;15:S43-S52doi: 10.1016/j.nephro.2019.03.008.
Rabant, M., Calvani, J., Terada, M., Lesaffre, C., Duong Van Huyen, J. P., & Bruneval, P. (2019). Immunofluorescence multiparamétrique in situ : vers l’amélioration du phénotype de l’infiltrat cellulaire au cours du rejet d’allogreffe rénale. In situ multiplex immunofluorescence analysis of the inflammatory burden in kidney allograft rejection: A new tool to characterize the alloimmune response. Nephrologie & therapeutique, 15S43-S52. https://doi.org/10.1016/j.nephro.2019.03.008
Rabant, Marion, et al. "Immunofluorescence multiparamétrique in situ : vers l’amélioration du phénotype de l’infiltrat cellulaire au cours du rejet d’allogreffe rénale." In situ multiplex immunofluorescence analysis of the inflammatory burden in kidney allograft rejection: A new tool to characterize the alloimmune response. Nephrologie & therapeutique vol. 15 (2019): S43-S52. doi: https://doi.org/10.1016/j.nephro.2019.03.008
Rabant M, Calvani J, Terada M, Lesaffre C, Duong Van Huyen JP, Bruneval P. Immunofluorescence multiparamétrique in situ : vers l’amélioration du phénotype de l’infiltrat cellulaire au cours du rejet d’allogreffe rénale. In situ multiplex immunofluorescence analysis of the inflammatory burden in kidney allograft rejection: A new tool to characterize the alloimmune response. Nephrol Ther. 2019 Apr;15:S43-S52. doi: 10.1016/j.nephro.2019.03.008. French. PMID: 30981395.
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Nephrol Ther. 2019 Apr;15:S43-S52. doi: 10.1016/j.nephro.2019.03.008.

In situ multiplex immunofluorescence analysis of the inflammatory burden in kidney allograft rejection: A new tool to characterize the alloimmune response.

Nephrologie & therapeutique

[Article in French]
Marion Rabant, Julien Calvani, Megumi Terada, Corinne Lesaffre, Jean-Paul Duong Van Huyen, Patrick Bruneval

Affiliations

  1. Laboratoire d'anatomie et cytologie pathologiques, hôpital Necker-Enfants-Malades, 149, rue de Sèvres, 75015 Paris, France; Inserm, U1151, institut Necker-Enfants-Malades, 149, rue de Sèvres, 75015 Paris, France; Université Paris Descartes, Sorbonne Paris Cité, 12, rue de l'École-de-Médecine, 75006 Paris, France; Paris Translational Research Center for Organ Transplantation, hôpital européen Georges-Pompidou, 20, rue Leblanc, 75015 Paris, France. Electronic address: [email protected].
  2. Laboratoire d'anatomie et cytologie pathologiques, hôpital Necker-Enfants-Malades, 149, rue de Sèvres, 75015 Paris, France; Inserm, UMR-S970, 20, rue Leblanc, 75015 Paris, France.
  3. Inserm, UMR-S970, 20, rue Leblanc, 75015 Paris, France; Laboratoire d'anatomie et cytologie pathologiques, hôpital européen Georges-Pompidou, 20 rue Leblanc, 75015 Paris, France.
  4. Inserm, UMR-S970, 20, rue Leblanc, 75015 Paris, France.
  5. Laboratoire d'anatomie et cytologie pathologiques, hôpital Necker-Enfants-Malades, 149, rue de Sèvres, 75015 Paris, France; Université Paris Descartes, Sorbonne Paris Cité, 12, rue de l'École-de-Médecine, 75006 Paris, France; Paris Translational Research Center for Organ Transplantation, hôpital européen Georges-Pompidou, 20, rue Leblanc, 75015 Paris, France; Inserm, UMR-S970, 20, rue Leblanc, 75015 Paris, France.
  6. Université Paris Descartes, Sorbonne Paris Cité, 12, rue de l'École-de-Médecine, 75006 Paris, France; Inserm, UMR-S970, 20, rue Leblanc, 75015 Paris, France; Laboratoire d'anatomie et cytologie pathologiques, hôpital européen Georges-Pompidou, 20 rue Leblanc, 75015 Paris, France.

PMID: 30981395 DOI: 10.1016/j.nephro.2019.03.008

Abstract

Background: The exact composition and localization of the inflammatory burden during allograft rejection is difficult to analyse on the same biopsy slide. We tested the feasibility of detecting four distinct markers in a same paraffin-embedded tissue section from human kidney allograft rejection by using an innovative process of multiplex immunofluorescence. Methods: Kidney allograft biopsies from 20 antibody-mediated rejection, 20 T cell-mediated rejection and five non rejection were labelled against NKp46, CD163, CD3, and CD34 respectively for NK cells, macrophages, T cells and endothelial cells. Images were scanned and cells were automatically quantified and their extra- or intravascular location determined. Conventional immunohistochemistry against NKp46 with manual quantification and real time quantitative polymerase chain reaction for evaluation of the relative messenger ribonucleic acid (mRNA) expression levels of NK, T cell and macrophage transcripts were simultaneously performed. Results: Multiplex immunofluorescence cell quantification was strongly correlated to manual quantification by immunohistochemistry (r = 0.91, P < 0.001) and to mRNA expression levels (r > 0.46, P < 0.021). T cells and macrophages were the two predominant populations involved in rejection (48.0 ± 4.4% and 49.3 ± 4.4% in antibody-mediated rejection; 51.8 ± 6.0% and 45.3 ± 5.8% in T cell-mediated rejection respectively) despite an important heterogeneity in the composition of the inflammatory burden. NK cells constituted a rare population for both T cell-mediated rejection (2.9 ± 0.6%) and antibody-mediated rejection (2.7 ± 0.7%). The intravascular compartment was mainly composed of T cells, including during antibody-mediated rejection. However, NK cells and macrophages densities were significantly higher in capillaries during antibody-mediated rejection. Conclusion: Multiplex immunofluorescence staining is a reliable technology allowing studying the exact composition and localization of the inflammatory burden during kidney allograft rejection..

Copyright © 2019. Published by Elsevier Masson SAS.

Keywords: Antibody mediated rejection; Cellules NK; Immunofluorescence multiparamétrique; Multiplex immunofluorescence; NK cells; Rejet cellulaire; Rejet humoral; T cell mediated rejection; Transplantation rénale

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