Abstract: FR-PO864
Multiplexed Immunofluorescence to Investigate the Immune Response to BK Virus in Kidney Transplant Recipients
Session Information
- Transplantation: Basic
October 26, 2018 | Location: Exhibit Hall, San Diego Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Transplantation
- 1801 Transplantation: Basic
Authors
- Fajardo, Cecile, Stanford University, Stanford, California, United States
- Grimm, Paul C., Stanford University Medical Center, Stanford, California, United States
- Kambham, Neeraja, Stanford University , Stanford, California, United States
Background
BK virus nephropathy is a complication in renal transplant that leads to allograft dysfunction or failure. Acute inflammation, tubular atrophy, and interstitial fibrosis are seen in both BKVN and acute rejection making it challenging to differentiate. Previous studies have characterized the immune response on histology, but the identification and characterization of infiltrating cells was limited due to constraints of traditional immunohistochemistry. Multiplexed immunofluorescence allows a more comprehensive understanding of the spatial relationships of immune cell phenotypes and ongoing signals within the kidney microenvironment.
Methods
Using CO-detection by inDEXing (CODEX), the expression of 50 markers for identifying renal structural elements, immune cell subsets, and immune function were evaluated on a single tissue section of a pediatric kidney transplant recipient.
Results
See images and caption for results.
Conclusion
CODEX is a novel technique in characterizing the immune repertoire in BKVN and visualizing renal histology. Predominant immune infiltrate in BKVN consists of T cells, plasmacytoid and myeloid dendritic cells, and sparse NK cells, which have not been previously described. A majority of CD3 T cells in BKVN appear to be activated and a minority expressing markers of exhaustion and inhibition. Comparing the immune repertoire of BKVN and acute rejection is an important and clinically significant future direction.
A) Renal tubules with cytokeratin, endothelial cells and blood vessels using CD31, and glomerular podocytes using podocin. B) Co-localization of CD3 T cells (green), CD1c myeloid dendritic cells (white), CD16 NK cells (yellow). C) Infiltration of interdigitating plasmacytoid dendritic cells (green) and myeloid dendritic cells (blue) in a cluster of T cells (red).
D) Functional analysis of CD3 T cells (blue) show co-expression of markers of inhibition with CD152/CTLA4 (green), exhaustion with CD279/PD1 (yellow), activation with CD69 (red), D4 shows a merge of all markers.