Analysis of Positive Flow Cytometric Crossmatch in Organ Transplantation

Yun Soo Lee; Dong Il Won

doi:10.3343/lmo.2011.1.1.7

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Lee and Won: Analysis of Positive Flow Cytometric Crossmatch in Organ Transplantation

Original Article

Diagnostic Immunology

Laboratory Medicine Online

Laboratory Medicine Online 2011; 1(1): 43-50.

Published online: 1 January 2011

DOI: https://doi.org/10.3343/lmo.2011.1.1.7

Analysis of Positive Flow Cytometric Crossmatch in Organ Transplantation

Yun Soo Lee, Dong Il Won

Department of Clinical Pathology, Kyungpook National University School of Medicine, Daegu, Korea.

Corresponding author: Yun Soo Lee. Department of Clinical Pathology, Kyungpook National University School of Medicine, 200 Dongdeong-ro, Jung-gu, Daegu 700-721, Korea. Tel: +82-53-420-5278, Fax: +82-53-426-3367, wormie@lycos.co.kr

Received 4 June 2010 Revised 4 October 2010 Accepted 5 October 2010

(open-access):

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

Pretransplant HLA crossmatch is one of the most important parts in solid organ transplantation. Flow cytometic crossmatch (FCXM) is more sensitive than anti-human globulin enhanced complement dependent lymphocytotoxicity (AHG-CDC) in detecting anti-HLA antibodies. We compared the results of the two methods and analyzed the FCXM-positive cases in various aspects.

Methods

Sera from 212 patients were tested for the detection of anti-HLA antibodies by FCXM and 188 of them were also tested by AHG-CDC assay. The results were analyzed in relation to their histories of pregnancy, transfusion or organ transplantation and also according to the donor patient relationships. We compared the FCXM results obtained before and after desensitization therapy (using plasmapheresis and anti-CD20 antibody) in 5 sensitized patients.

Results

Concordance of the results between the two methods was 88.8% (167/188). FCXM results correlated with history of pregnancy, but not with that of transfusion. When the patients were divided into 4 groups according to donor-patient relationships, the T cell FCXM mean fluorescence intensity (MFI) ratio (sample/control) was significantly higher in the husband-to-multiparous wife group compared to the other 3 groups (children-to-mother, unrelated donor-to-multipara, and the rests). After desensitization therapy, MFI ratios of T cell FCXM decreased and those of B cell FCXM increased, probably due to rituximab effect, in all 5 patients.

Conclusions

FCXM using a MFI ratio, has a higher sensitivity than AHG-CDC in detection of donor specific antibodies. Also it can be useful in monitoring antibody levels during desensitization therapy.

Keywords: Flow cytometric crossmatch, Anti-HLA antibodies, Organ transplantation

Figures and Tables

Fig. 1

Desensitization therapy protocols. (A) In patients 1 and 2 of Table 7. (B) In patients 3, 4 and 5 of Table 7.

Abbreviations: PP, plasmapheresis; KT, kidney transplantation.

Fig. 2

Distribution of the mean fluorescence intensity (MFI) ratios in each relationship of donor and patient. (A) T cell MFI ratios. (B) B cell MFI ratios. The mean is indicated.

^*the ANOVA with a subsequent LSD post hoc test.

Table 1

Relationships of donor and patient

Table 2

Comparison of results between T cell/B cell FCXM and AHG-CDC

^*positive B cell FCXM; ^†positive T cell FCXM.

Abbreviations: FCXM, flow cytometric crossmatch; AHG-CDC, anti-human immunoglobulin- complement dependent (lympho)cytotoxicity.

Table 3

FCXM results in female patients in relation to pregnancy history: transfusion history (-)/transfusion history (+)/all patients

^*excluded 19 cases with pregnancy information missing; ^†Chi-square test.

Abbreviations: See Table 2.

Table 4

FCXM results in female patients in relation to transfusion history: pregnancy history (-)/pregnancy history (+)/all patients

^*excluded 19 cases with pregnancy information missing; ^†Chi-square test.

Abbreviations: See Table 2.

Table 5

FCXM results in male patients in relation to transfusion history

^*Chi-square test.

Abbreviations: See Table 2.

Table 6

Characteristics of patients who had a history of organ transplantation

Abbreviations: See Table 2.

Table 7

Changes in MFI ratio in patients who received desensitization therapy

^*protocol (A) of Fig. 1. in patient 1 and 2, (B) in 3, 4 and 5; ^†day before transplantation.

Abbreviations: MFI, mean fluorescence intensity; See Table 2.

Notes

This article is available from http://www.labmedonline.org

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