Flow Cytometric AHG-CDC for HLA Crossmatch: A Pilot Study.
- Author:
Dong Il WON
1
Author Information
1. Department of Laboratory Medicine, Kyungpook National University Hospital, Daegu, Korea. wondi@knu.ac.kr
- Publication Type:Original Article
- Keywords:
HLA crossmatch;
AHG-CDC;
Flow cytometry
- MeSH:
Antibodies;
B-Lymphocytes;
Centers for Disease Control and Prevention (U.S.);
Complement System Proteins;
Dactinomycin;
Flow Cytometry;
Organ Transplantation;
Pilot Projects;
T-Lymphocytes;
Transplants
- From:Korean Journal of Blood Transfusion
2010;21(2):122-131
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: For HLA crossmatch in organ transplantation, complement-dependent cytotoxicity (CDC) is a very useful methodology to detect donor-specific HLA antibodies and their complement fixing ability. In this preliminary pilot study, we investigated whether dead cells induced in anti-human globulin (AHG)-augmented CDC (AHG-CDC) reaction could be measured by flow cytometry ('FC AHG-CDC'). METHODS: This FC AHG-CDC measured the percentage of dead cells (% dead cells) after 7-aminoactinomycin D staining using 3 color flow cytometry. A total 45 flow cytometry crossmatch (FCXM) cases of 12 positives and 33 negatives were further tested using this FC AHG-CDC. RESULTS: The % dead cells of FC AHG-CDC was significantly correlated with the mean fluorescent intensity ratio of FCXM (T cells: r=0.613, P=7.45x10(-6); B cells: r=0.404, P=0.006). The positivity rate of FC AHG-CDC among FCXM positive cases was relatively high: 80% (8/10) for T cells and 75% (9/12) for B cells. The negativity rate of FC AHG-CDC among FCXM negative cases was 100% (35/35) for T cells and 91% (30/33) for B cells. CONCLUSION: In this pilot study, FC AHG-CDC could yield quantitative values, % dead cells, which was proportional to the level of complement-fixing cytotoxic antibodies against T and B cells, respectively, even without physical separation of cells or serial dilution of serum.
