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Updated: Nov 4, 2025
Personalized Peptide Arrays for Detection of HLA Alloantibodies in Organ Transplantation
Published on: September 6, 2017
Improved detection of donor-specific HLA-class II antibody in kidney transplant recipients by modified immunocomplex
Shuichi Shimabukuro1, Kenta Iwasaki2, Shintaro Kawai3
1Department of Urology, Okinawa Chubu Hospital, 281, Miyazato, Uruma, Okinawa, Japan; Department of Renal Transplant Surgery, Aichi Medical University School of Medicine, Nagakute, Japan.
Insights
Modified Immunocomplex Capture Fluorescence Analysis (ICFA) using peripheral blood mononuclear cells (PBMCs) enhances donor-specific HLA antibody (DSA) detection for kidney transplant recipients. This optimized ICFA-PBMC method improves accuracy for HLA-class II antibody detection, aiding in pre-transplant and post-transplant assessments.
Area of Science:
- Transplantation immunology
- Immunodiagnostics
- HLA antibody detection
Background:
- Donor-specific HLA antibodies (DSA) are critical in kidney transplant rejection.
- Flow cytometry crossmatch (FCXM) is sensitive but requires viable cells.
- Immunocomplex capture fluorescence analysis (ICFA) detects DSA without viable cells but has lower sensitivity, especially for HLA-class II.
Purpose of the Study:
- To investigate and improve the accuracy of ICFA for detecting HLA-class II DSA.
- To enhance the sensitivity of ICFA for HLA-class II antibody detection.
- To evaluate the performance of an optimized ICFA method in kidney transplant patients.
Main Methods:
- Modified ICFA using purified peripheral blood mononuclear cells (PBMCs) instead of whole blood.
- Compared ICFA-PBMC with FCXM-T/B in 118 pre-transplant and 13 post-transplant patients.
- Determined optimal cutoff level using receiver operating characteristic curve analysis.
Main Results:
- A significant positive correlation was found between ICFA-PBMC values and HLA-class II DSA mean fluorescence intensity (MFI).
- The optimized ICFA-PBMC method demonstrated significantly higher average DSA MFI in positive cases compared to negative cases (12,217 vs 3885).
- An optimized cutoff level of 1.4 was established for ICFA-PBMC.
Conclusions:
- Modified ICFA using PBMCs (ICFA-PBMC) significantly improves sensitivity for HLA-class II DSA detection.
- ICFA-PBMC with an optimized cutoff provides valid information for suspected DSA, complementing existing methods.
- This enhanced ICFA method offers a valuable tool for assessing HLA-class II antibodies in transplantation.
Abstract:
Immunocomplex capture fluorescence analysis (ICFA) which basic principle is same as Luminex crossmatch (LXM), could detect donor-specific HLA antibody (DSA). The advantages of ICFA are (i) detection of DSA and (ii) no requirement of viable cells over the flow cytometry crossmatch (FCXM). However, FCXM has been widely used because of its higher sensitivity than ICFA, in particular HLA-class II antibody detection. In this study the accuracy of DSA detection against HLA-class II was investigated by modifying the original method of ICFA. Increment of the sensitivity was found when purified peripheral blood mononuclear cells (PBMCs) were used instead of whole blood. An ICFA-PBMC in addition to FCXM-T/B was conducted for 118 patients before kidney transplantation and 13 patients with de novo DSA against HLA-class II after transplantation. Significantly positive correlation was observed between the values of ICFA-PBMC and DSA mean fluorescence intensity (MFI) targeting class II (p < 0.0001). When the cutoff level of 1.4 was determined by receiver operating characteristic curve analysis, the average DSA MFI was found to be significantly higher in the ICFA-PBMC (class II) positive group comparing to that in the negative group (12,217 vs 3885, p = 0.0027). ICFA-PBMC and optimized cutoff level could provide valid information in cases of suspected DSA.
