EDTA Treatment for Overcoming the Prozone Effect and for Predicting C1q Binding in HLA Antibody Testing
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The Luminex-based single antigen bead (SAB) assay is widely used to detect HLA antibody in transplant recipients. However, one limitation of the SAB assay is the prozone effect, which occurs mostly as a result of complement interference. We investigated the efficacy of EDTA treatment for overcoming the prozone effect and predicting C1q binding of HLA antibody. We subjected 27 non-treated (naïve) and EDTA-treated serum samples from highly sensitized patients to IgG-SAB assays, and we confirmed the prozone effect in 53% and 31% of class I and class II antibody tests, respectively, after EDTA treatment. When we conducted additional assays after dithiothreitol treatment and serum dilution, EDTA was the most efficacious in eliminating the prozone effect. Reducing the prozone effect by EDTA treatment strengthened the correlation between IgG mean fluorescence intensity (MFI) and C1q MFI values (ρ=0.825) as compared with the naïve sera (ρ=0.068). Although C1q positivity was dependent on the concentration of HLA antibody in EDTA-treated sera, the correlations varied individually. Overall, our results confirmed the efficacy of EDTA treatment for overcoming the prozone effect. EDTA treatment showed a positive effect on the correlation between IgG MFI and C1q MFI values.
Abstract
The Luminex-based single antigen bead (SAB) assay is widely used to detect HLA antibody in transplant recipients. However, one limitation of the SAB assay is the prozone effect, which occurs mostly as a result of complement interference. We investigated the efficacy of EDTA treatment for overcoming the prozone effect and predicting C1q binding of HLA antibody. We subjected 27 non-treated (naïve) and EDTA-treated serum samples from highly sensitized patients to IgG-SAB assays, and we confirmed the prozone effect in 53% and 31% of class I and class II antibody tests, respectively, after EDTA treatment. When we conducted additional assays after dithiothreitol treatment and serum dilution, EDTA was the most efficacious in eliminating the prozone effect. Reducing the prozone effect by EDTA treatment strengthened the correlation between IgG mean fluorescence intensity (MFI) and C1q MFI values (ρ=0.825) as compared with the naïve sera (ρ=0.068). Although C1q positivity was dependent on the concentration of HLA antibody in EDTA-treated sera, the correlations varied individually. Overall, our results confirmed the efficacy of EDTA treatment for overcoming the prozone effect. EDTA treatment showed a positive effect on the correlation between IgG MFI and C1q MFI values.
참고문헌 (20)
- Tambur AR, "Sensitization in transplantation : assessment of risk(STAR)2017 working group meeting report", Am J Transplant, 18, pp.1604~1614 (2018)
- 오은지, "Interlaboratory Comparison of the Results of Lifecodes LSA Class I and Class II Single Antigen Kits for Human Leukocyte Antigen Antibody Detection", Annals of Laboratory Medicine, 35(3), pp.321~328 (2015), 대한진단검사의학회
- Gebel HM, "The evolution and clinical impact of human leukocyte antigen technology", Curr Opin Nephrol Hypertens, 19, pp.598~602 (2010)
- 장지영, "Application of Calculated Panel Reactive Antibody Using HLA Frequencies in Koreans", Annals of Laboratory Medicine, 32(1), pp.66~72 (2012), 대한진단검사의학회
- 민지원, "Clinical Impact of Pre-transplant Antibodies Against Angiotensin II Type I Receptor and Major Histocompatibility Complex Class I-Related Chain A in Kidney Transplant Patients", Annals of Laboratory Medicine, 38(5), pp.450~457 (2018), 대한진단검사의학회[DOI]
- Mujtaba MA, "The strength of donor-specific antibody is a more reliable predictor of antibody-mediated rejection than flow cytometry crossmatch analysis in desensitized kidney recipients", Clin Transplant, 25, pp.E96~E102 (2011)
- Zachary A, "Quantifying HLA-specific antibodies in patients undergoing desensitization", Curr Opin Organ Transplant, 16, pp.410~415 (2011)
- Lee H, "Clinical impact of complement(C1q, C3d)binding de novo donor-specific HLA antibody in kidney transplant recipients", PLoS One, 13, pp.e0207434~ (2018)
- Tambur AR, "HLA diagnostics : evaluating DSA strength by titration", Transplantation, 102, pp.S23~S30 (2018)[DOI]
- Tambur AR, "Assessing antibody strength : comparison of MFI, C1q, and titer information", Am J Transplant, 15, pp.2421~2430 (2015)
- Schnaidt M, "HLA antibody specification using single-antigen beads—a technical solution for the prozone effect", Transplantation, 92, pp.510~515 (2011)
- Zhang X, "Comprehensive assessment for serum treatment for single antigen test for detection of HLA antibodies", Hum Immunol, 78, pp.699~703 (2017)
- Ellis TM, "Interpretation of HLA single antigen bead assays", Transplant Rev(Orlando), 27, pp.108~111 (2013)
- Tambur AR, "Can solid phase assays be better utilized to measure efficacy of antibody removal therapies", Hum Immunol, 77, pp.624~630 (2016)
- Kosmoliaptsis V, "Detection of immunoglobulin G human leukocyte antigen-specific alloantibodies in renal transplant patients using single-antigen-beads is compromised by the presence of immunoglobulin M human leukocyte antigen-specific alloantibodies", Transplantation, 87, pp.813~820 (2009)
- Schwaiger E, "Complement component C3 activation : the leading cause of the prozone phenomenon affecting HLA antibody detection on single-antigen beads", Transplantation, 97, pp.1279~1285 (2014)
- Liu C, "Quantitative evaluation of the impact of ethylenediaminetetraacetic acid pretreatment on single-antigen bead assay", Transplant Direct, 3, pp.e194~ (2017)
- Guidicelli G, "Prevalence, distribution and amplitude of the complement interference phenomenon in single antigen flow beads assays", HLA, 91, pp.507~513 (2018)
- Hahn AB, "ASHI laboratory manual", pp.4.1~ (2000), Society for Histocompatibility and Immunogenetics
- Taylor CJ, "Technical limitations of the C1q Single-Antigen Bead Assay to detect complement binding HLA-specific antibodies", Transplantation, 101, pp.1206~1214 (2017)