Chronic Active Antibody-Mediated Rejection Is Associated With the Upregulation of Interstitial But Not Glomerular Transcripts.

Andriy Trailin,Matej Kocik,Petra Mrazova,Eva Sticova,Ondrej Viklicky,Antonij Slavcev,Petra Hruba,Marek Novotny,Ludek Voska

doi:10.3389/fimmu.2021.729558

Abstract

Molecular assessment of renal allografts has already been suggested in antibody-mediated rejection (ABMR), but little is known about the gene transcript patterns in particular renal compartments. We used laser capture microdissection coupled with quantitative RT-PCR to distinguish the transcript patterns in the glomeruli and tubulointerstitium of kidney allografts in sensitized retransplant recipients at high risk of ABMR. The expressions of 13 genes were quantified in biopsies with acute active ABMR, chronic active ABMR, acute tubular necrosis (ATN), and normal findings. The transcripts were either compartment specific (TGFB1 in the glomeruli and HAVCR1 and IGHG1 in the tubulointerstitium), ABMR specific (GNLY), or follow-up specific (CXCL10 and CX3CR1). The transcriptional profiles of early acute ABMR shared similarities with ATN. The transcripts of CXCL10 and TGFB1 increased in the glomeruli in both acute ABMR and chronic active ABMR. Chronic active ABMR was associated with the upregulation of most genes (SH2D1B, CX3CR1, IGHG1, MS4A1, C5, CD46, and TGFB1) in the tubulointerstitium. In this study, we show distinct gene expression patterns in specific renal compartments reflecting cellular infiltration observed by conventional histology. In comparison with active ABMR, chronic active ABMR is associated with increased transcripts of tubulointerstitial origin.

Highlights

Both acute antibody-mediated rejection (ABMR) and chronic ABMR are the main risk factors for late renal allograft loss [1,2,3]
The gene expression patterns in inflamed kidneys during various kidney diseases or allograft rejection are supposed to correspond to the cellular infiltration observed in conventional histology
Despite current achievements in molecular diagnostics in kidney transplantation, little is known about renal compartment gene expression patterns in transplant pathologies

Summary

INTRODUCTION

Both acute antibody-mediated rejection (ABMR) and chronic ABMR are the main risk factors for late renal allograft loss [1,2,3]. We used LCM to examine the compartmentspecific expression patterns of selected genes in sensitized kidney transplant recipients who suffered from active and chronic active ABMR. A small portion (~2 mm) of all for-cause and 3-month (3M) protocol biopsies, performed from January 2015 until January 2019 in sensitized kidney transplant recipients who had undergone a second, third, or a fourth kidney transplantation, were embedded into Tissue-Tek® O.C.T. Compound (Sakura Finetek USA, Inc., Torrance, CA, USA), snap frozen in liquid nitrogen immediately, and stored at −80°C for future LCM and gene expression analyses (N = 211). Samples with pure diagnosis of active ABMR, chronic active ABMR, ATN and normal findings in 3-month protocol biopsy, which had enough material for LCM and good RNA quality, were used for RT-qPCR form the biobank of biopsies from retransplanted patients. ABMR, antibody-mediated rejection; PRA, panel-reactive antibodies; HLA, human leukocyte antigen; DSA, donor-specific antibodies; MICA, major histocompatibility complex class I chain-related antigen A; ATG, anti-thymocyte globulin; D, deceased; ECD, expanded criteria donor; CIT, cold ischemia time. aData are presented as median (minimum–maximum). bTotal time on dialysis. cAt the time of biopsy

RESULTS

DISCUSSION

ETHICS STATEMENT