Graft Immunocomplex Capture Fluorescence Analysis to Detect Donor Specific Antibodies and HLA Antigen Complexes in the Renal Allograft.
1Department of Organ Transplantation and General Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan
2Department of Blood Transfusion and Cell Therapy, Kyoto Prefectural University of Medicine, Kyoto, Japan
Meeting: 2017 American Transplant Congress
Abstract number: A9
Keywords: Antibodies
Session Information
Session Name: Poster Session A: Antibody Mediated Rejection in Kidney Transplant Recipients I
Session Type: Poster Session
Date: Saturday, April 29, 2017
Session Time: 5:30pm-7:30pm
Presentation Time: 5:30pm-7:30pm
Location: Hall D1
Background: Immunocomplex Capture Fluorescence Analysis (ICFA) is an attractive method to detect donor specific anti-HLA antibodies (DSA) and HLA antigen complexes. Currently, antibody-mediated rejection (AMR) due to DSA is usually diagnosed by microvasculitis, C4d deposition and serological DSA detection in renal transplantation. Conversely, there is a discrepancy between these findings frequently. Thereupon, graft ICFA technique may contribute to establish the diagnosis of AMR.
Methods:We enrolled 40 consecutive renal transplant recipients indicated for graft biopsies due to various reasons. Samples were obtained by a percutaneous needle biopsy. Then, the specimen was dissolved in PBS by the lysis buffer. Subsequently, HLA antigens were captured by anti HLA beads. Then, DSA-HLA complexes were detected by PE-conjugated anti human IgG antibodies, where DSA had already reacted with the allograft in vivo, analyzed by a Luminex system. 
Results: A ratio (sample/blank beads MFI) was calculated: ≥ 1.0 was determined as positive. The mean negative result was 0.5 ± 0.2. DSA-HLA complexes in the graft were successfully detected from slight positive 1.03, to 79.27 in a chronic active AMR patient. In, preformed DSA positive recipients, g-DSA deposition can be confirmed (1.15 ± 0.04) at 1 hour after re-perfusion. Subsequently, g-DSA shifted to (2.20 ±0.98) at 3 weeks following transplantation along with a decline in s-DSA MFI: 1,718 to 506.5. Next, positive graft ICFA had predicted the early phase of AMR (MFI ratio: 1.38) even in patients with no serum DSA. Finally, appropriate therapies for AMR deleted DSA deposition (0.3 to 0.7) from allografts.
Conclusions: This novel application would detect early phase or incomplete pathological cases of AMR, which could lead to a correct diagnosis and initiation of appropriate therapies. Moreover, graft ICFA might address a variety of long standing questions in terms of DSA.
CITATION INFORMATION: Nakamura T, Ushigome H, Watabe K, Imanishi Y, Harada S, Nobori S, Masuda K, Yoshimura N. Graft Immunocomplex Capture Fluorescence Analysis to Detect Donor Specific Antibodies and HLA Antigen Complexes in the Renal Allograft. Am J Transplant. 2017;17 (suppl 3).
To cite this abstract in AMA style:
Nakamura T, Ushigome H, Watabe K, Imanishi Y, Harada S, Nobori S, Masuda K, Yoshimura N. Graft Immunocomplex Capture Fluorescence Analysis to Detect Donor Specific Antibodies and HLA Antigen Complexes in the Renal Allograft. [abstract]. Am J Transplant. 2017; 17 (suppl 3). https://atcmeetingabstracts.com/abstract/graft-immunocomplex-capture-fluorescence-analysis-to-detect-donor-specific-antibodies-and-hla-antigen-complexes-in-the-renal-allograft/. Accessed May 18, 2025.« Back to 2017 American Transplant Congress