Evaluation of Flow Cytometric Crossmatch Results in Comparison with Donor-specific Antibodies Detected by Luminex-PRA Tests in Organ Transplantation Patients

Seon Young Kim; Bok Youn Han; Jungwon Hyun; Shin Young Joo; Eun Young Song; Myoung Hee Park

doi:10.4285/jkstn.2012.26.2.92

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Kim, Han, Hyun, Joo, Song, and Park: Evaluation of Flow Cytometric Crossmatch Results in Comparison with Donor-specific Antibodies Detected by Luminex-PRA Tests in Organ Transplantation Patients

Original Article

The Journal of the Korean Society for Transplantation

The Journal of the Korean Society for Transplantation 2012; 26(2): 92-100.

Published online: 29 June 2012

DOI: https://doi.org/10.4285/jkstn.2012.26.2.92

Evaluation of Flow Cytometric Crossmatch Results in Comparison with Donor-specific Antibodies Detected by Luminex-PRA Tests in Organ Transplantation Patients

Seon Young Kim, M.D.¹, Bok Youn Han, M.T.¹, Jungwon Hyun, M.D.², Shin Young Joo, M.D.³, Eun Young Song, M.D.¹, Myoung Hee Park, M.D.¹

¹Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.

²Department of Laboratory Medicine, Hallym University Sacred Heart Hospital, Anyang, Korea.

³Department of Laboratory Medicine, Seoul Paik Hospital, Inje University College of Medicine, Seoul, Korea.

Corresponding author (parkmhee@snu.ac.kr)

Received 31 October 2011 Accepted 2 May 2012

Abstract

Background

Two of the most sensitive methods for detecting donor-specific HLA antibodies (DSAs) are solid phase panel reactive antibody (PRA) assay using Luminex platform (Luminex-PRA), and a cell-based flow cytometric crossmatch (FCXM) test. We evaluated FCXM results in relation to DSAs detected by the Luminex-PRA method in solid organ transplantation candidates or post-transplant follow-up patients.

Methods

A total of 171 donor-recipient pairs were evaluated by Luminex-PRA (LIFECODES Class I and Class II ID kits; Gen-Probe, USA) and FCXM (T- and B-cells) tests. DSA levels were analyzed using a sum of median fluorescence intensity (MFI) values, and FCXM results were analyzed using MFI ratios.

Results

Class I and II DSAs were detected in 11.7% (20/171) and 11.1% (19/171) of tested sera, respectively. T-FCXM was negative in 97.4% (147/151) of Class I DSA negative sera, and B-FCXM was negative in 99.3% (137/138) of Class I and II DSA negative sera. T-FCXM was positive in 91.7% (11/12) of sera with moderate to strong Class I DSAs and B-FCXM was positive in 88.9% (16/18) of sera with moderate to strong Class II and/or Class I DSAs in the evaluation of sensitivities of FCXM in relation to DSA. There were significant correlations between FCXM ratios and DSA levels for both T-FCXM (P=0.008) and B-FCXM (P<0.001).

Conclusions

The FCXM results correlated well with the DSAs detected by the Luminex-PRA method. The specificities of T- and B-FCXM in relation to DSAs were high (>97%) and the sensitivities of T- and B-FCXM were satisfactory (>88%) in detecting moderate to strong DSAs.

Keywords: HLA antigens, Panel reactive antibody, Histocompatibility testing, Flow cytometry, Crossmatching

Figures and Tables

Fig. 1

Relationship between DSA intensity (MFI sum) and FCXM strength (MFI ratio). (A) T-FCXM results in patients with Class I DSA (n=20), and (B) B-FCXM results in patients with Class I or Class II DSA (n=33). Abbreviations: DSA, donor-specific antibody; FCXM, flow cytometric crossmatch; MFI, median fluorescence intensity.

Fig. 2

Relationship between DSA intensity and flow cytometric crossmatch (FCXM) results. Sensitivities of FCXM tests detecting DSA-positive samples of different intensities are shown. Abbreviations: DSA, donor-specific antibody; MFI, median fluorescence intensity.

Fig. 3

Receiver operating characteristics (ROC) curve analysis of DSA intensity (MFI sum) for FCXM positivity. MFI sum values of Class I DSAs were used for T-FCXM (A), and those of Class I and Class II were used for B-FCXM (B). Abbreviations: AUC, area under the curve; DSA, donor-specific antibody; FCXM, flow cytometric crossmatch; MFI, median fluorescence intensity.

Table 1

Results of Luminex-PRA and flow cytometric crossmatch (FCXM) tests

Abbreviations: PRA, panel reactive antibody; DSA, donor-specific antibody.

^aLuminex-PRA results in 156 samples from 141 patients, and DSA and FCXM results in 171 donor-recipient pairs; ^bResults of Luminex-PRA screening test for HLA Class I and Class II antibodies; ^cDSA detected by Luminex-PRA test.

Table 2

Comparison between the results of Luminex-PRA and flow cytometric crossmatch (FCXM) tests

Abbreviations: PRA, panel reactive antibody; DSA, donor-specific antibody.

Table 3

Flow cytometric crossmatch (FCXM) results according to DSA intensity

Abbreviations: DSA, donor-specific antibody; MFI, median fluorescence intensity; n, number.

^aSum of MFI of Class I DSA was calculated for T-FCXM, and sum of MFI of Class I and ClassII DSA was calculated for B-FCXM; ^bMedian value with ranges in parentheses; ^cP values by Kruskal-Wallis ANOVA test.

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