In situ multiplex immunofluorescence analysis of the inflammatory burden in kidney allograft rejection: A new tool to characterize the alloimmune response. Issue 4 (11th December 2019)

• Record Type: Journal Article
• Authors: Calvani, Julien; Terada, Megumi; Lesaffre, Corinne; Eloudzeri, Maëva; Lamarthée, Baptiste; Burger, Carole; Tinel, Claire; Anglicheau, Dany; Vermorel, Agathe; Couzi, Lionel; Loupy, Alexandre; Duong Van Huyen, Jean‐Paul; Bruneval, Patrick; Rabant, Marion
• Is Part Of: American journal of transplantation. Volume 20:Issue 4(2020)
• Page Start: 942
• Page End: 953
• Publication Date: 2019-12-11
• Subjects: immunohistochemistry -- kidney transplantation/nephrology -- natural killer (NK) cells/NK receptors -- pathology/histopathology -- rejection -- translational research/science; Transplantation of organs, tissues, etc -- Periodicals; 617.95
• DOI: 10.1111/ajt.15699
• Languages: English
• ISSNs: 1600-6135

Abstract

Abstract : The exact composition of leukocyte infiltration during kidney allograft rejection is difficult to comprehend and visualize on the same biopsy slide. Using an innovative technology of multiplex immunofluorescence (mIF), we were able to detect simultaneously NK cells, macrophages, and T cells and to determine their intra‐ or extravascular localization using an endothelial marker. Twenty antibody‐mediated rejection (ABMR), 20 T cell–mediated rejection (TCMR), and five normal biopsies were labeled, with automatic leukocyte quantification and localization. This method was compared to a classic NKp46 immunohistochemistry (IHC) with manual quantification and to mRNA quantification. mIF automatic quantification was strongly correlated to IHC ( r = .91, P < .001) and to mRNA expression levels ( r > .46, P < .021). T cells and macrophages were the 2 predominant populations involved in rejection (48.0 ± 4.4% and 49.3 ± 4.4%, respectively, in ABMR; 51.8 ± 6.0% and 45.3 ± 5.8% in TCMR). NK cells constituted a rare population in both ABMR (2.7 ± 0.7%) and TCMR (2.9 ± 0.6%). The intravascular compartment was mainly composed of T cells, including during ABMR, in peritubular and glomerular capillaries. However, NK cell and macrophage densities were significantly higher during ABMR in glomerular and peritubular capillaries. To conclude, this study demonstrates the feasibility and utility of mIF imaging to study and better understand the kidney allograft rejection process. Abstract : In this study, the authors use an innovative technology of multiplexed immunofluorescence on paraffin‐embedded biopsies to detect simultaneously on the same slide NK cells, macrophages, and T cells and to determine their intra‐ or extravascular localization during T cell‐ and antibody‐mediated rejection. See editorial by Bellamy and Prost on page 915 .