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Graft Immunocomplex Capture Fluorescence Analysis to Detect Donor-Specific Antibodies and HLA Antigen Complexes in the Allograft
- Source :
- Immunological investigations. 46(3)
- Publication Year :
- 2017
-
Abstract
- 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 C4d deposition and serological DSA detection. Conversely, there is a discrepancy between these findings frequently. Thereupon, our graft ICFA technique may contribute to establish the diagnosis of AMR.Graft 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.A ratio (sample MFI/blank beads MFI) was calculated: ≥ 1.0 was determined as positive. We found that DSA-HLA complexes in the graft were successfully detected from only slight positive 1.03 to 79.27 in a chronic active AMR patient by graft ICFA. 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 (MFI ratio from 0.3 to 0.7) from allografts.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.AMR: Antibody-mediated rejection; DSA: Donor-specific antibodies; ICFA: Immunocomplex capture fluorescence analysis.
- Subjects :
- Adult
Graft Rejection
Male
Pathology
medicine.medical_specialty
Immunology
Fluorescent Antibody Technique
Human leukocyte antigen
Antigen-Antibody Complex
030230 surgery
Kidney
Serology
03 medical and health sciences
0302 clinical medicine
In vivo
HLA Antigens
Isoantibodies
Lysis buffer
medicine
Humans
Aged
biology
Percutaneous needle biopsy
Chemistry
Donor specific antibodies
Antibody-Dependent Cell Cytotoxicity
General Medicine
Middle Aged
Allografts
Fluorescence
Molecular biology
Kidney Transplantation
body regions
biology.protein
Female
Antibody
030215 immunology
Subjects
Details
- ISSN :
- 15324311
- Volume :
- 46
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Immunological investigations
- Accession number :
- edsair.doi.dedup.....2d7e5177ddf7290862f3732a563b63af