Evaluation of lymphocyte crossmatch (CDCXM and Flow XM) test results before kidney transplantation

Öz

Purpose: Detection of anti-HLA antibodies is crucial in pre-transplant immunological assessment to evaluate donor-recipient compatibility and prevent hyperacute rejection. This study aimed to evaluate lymphocyte crossmatch (XM) test results performed before kidney transplantation.
Materials and methods: Lymphocyte crossmatch tests conducted at the Tissue Typing Laboratory of Pamukkale University Hospital between February 2020 and February 2024 were retrospectively analyzed. CDCXM (Complement-Dependent Cytotoxicity Crossmatch) and flow cytometric crossmatch (FCXM) tests were performed. For CDCXM, T and B lymphocytes were separated and tested individually, with serial dilution and DTT treatment applied. FCXM used anti-CD3 for T lymphocytes, anti-CD19 and anti-IgG for B lymphocytes, analyzed on a 10-color, 3-laser flow cytometer (Navios EX-Beckman Coulter, Inc. USA).
Results: A total of 225 CDCXM test results, 105 of which were alive and 120 of which were cadaver, were included in the study. CDCXM T and FCXM T were negative in living donors, while CDCXM B and FCXM B positivity were 4.8% and 2.6%, respectively. CDCXM T and B positivity in cadaveric donors were 3.3% and 7.5%, respectively, and FCXM T and B positivity were 3.3% and 8.8%, respectively.
Conclusion: Lymphocyte crossmatch remains a reliable and classical method for pre-transplant immunological evaluation. It effectively detects donor-specific anti-HLA antibodies and contributes to the prevention of hyperacute rejection in kidney transplantation.

Anahtar Kelimeler

• Kidney transplantation
• lymphocyte crossmatch
• CDC crossmatch
• flowcytometric crossmatch
• anti-HLA antibodies

Böbrek nakli öncesi yapılan lenfosit crossmatch (CDCXM ve Flow XM) test sonuçlarının değerlendirilmesi

Öz

Amaç: Anti-HLA antikorlarının tespiti, donör-alıcı uyumunu değerlendirmek ve hiperakut reddi önlemek için nakil öncesi immünolojik değerlendirmede kritik öneme sahiptir. Bu çalışmada, böbrek nakli öncesinde yapılan lenfosit çapraz uyum (XM) testi sonuçlarının değerlendirilmesi amaçlanmıştır.
Gereç ve yöntem: Şubat 2020 ile Şubat 2024 tarihleri arasında Pamukkale Üniversitesi Hastanesi Doku Tipleme Laboratuvarı'nda gerçekleştirilen lenfosit çapraz uyum testleri retrospektif olarak analiz edilmiştir. CDCXM (Komplement Bağımlı Sitotoksisite Çapraz Uyum) ve akım sitometrik çapraz uyum (FCXM) testleri gerçekleştirilmiştir. CDCXM için T ve B lenfositleri ayrı ayrı ayrı test edilmiş, seri dilüsyon ve DTT uygulanmıştır. FCXM'de T lenfositleri için anti-CD3, B lenfositleri için anti-CD19 ve anti-IgG kullanılmış ve 10 renkli, 3 lazerli akım sitometresinde (Navios EX-Beckman Coulter, Inc. USA) analiz edilmiştir.
Bulgular: Çalışmaya 105’i canlı ve 120’si kadavrada olmak üzere toplam Toplam 225 CDCXM test sonucu dahil edilmiştir. Canlı donörlerde CDCXM T ve FCXM T negatif bulunurken, CDCXM B ve FCXM B pozitifliği sırasıyla %4,8 ve %2,6 idi. Kadavra donörlerde CDCXM T ve B pozitifliği sırasıyla %3,3 ve %7,5, FCXM T ve B pozitifliği ise sırasıyla %3,3 ve %8,8 idi.
Sonuç: Lenfosit çapraz eşleştirme, nakil öncesi immünolojik değerlendirme için güvenilir ve klasik bir yöntem olmaya devam etmektedir. Vericiye özgü anti-HLA antikorlarını etkili bir şekilde tespit eder ve böbrek naklinde hiperakut reddin önlenmesine katkıda bulunur.

Anahtar Kelimeler

• Böbrek nakli
• lenfosit çapraz eşleştirme
• CDC çapraz eşleştirme
• flowsitometrik çapraz eşleştirme
• anti-HLA antikorları

Kaynakça

1. Naik RH, Shawar SH. Acute Renal Transplantation Rejection. 2023 Feb 9. In: StatPearls [Internet]. Treasure Island (FL): StatPearls.
2. Garcia GG, Harden P, Chapman J; World Kidney Day Steering Committee 2012. The global role of kidney transplantation. Kidney Blood Press Res. 2012;35(5):299-304. doi:10.1159/000337044
3. Saito PK, Yamakawa RH, Pereira LC, da Silva WV Jr, Borelli SD. Complement-dependent cytotoxicity (CDC) to detect Anti-HLA antibodies: old but gold. J Clin Lab Anal. 2014;28(4):275-280. doi:10.1002/jcla.21678
4. Jayant K, Reccia I, Julie BM, Sharma A, Halawa A. Role of crossmatch testing when Luminex-SAB is negative in renal transplantation. Pol Przegl Chir. 2018;90(1):41-46. doi:10.5604/01.3001.0011.5959
5. Gebel HM, Bray RA, Nickerson P. Pre-transplant assessment of donor-reactive, HLA-specific antibodies in renal transplantation: contraindication vs. risk. Am J Transplant. 2003;3(12):1488-1500. doi:10.1046/j.1600-6135.2003.00273.x
6. Terasaki PI, McClelland JD. Microdroplet Assay of Human Serum Cytotoxins. Nature. 1964;204:998-1000. doi:10.1038/204998b0
7. Mahowald GK. The CDC crossmatch in the era of flow cytometric cross-match and single antigen beads. J Bras Nefrol. 2021;43(3):299-300. doi:10.1590/2175-8239-JBN-2021-0110
8. Focosi D. Advances in Pretransplant Donor-Specific Antibody Testing in Solid Organ Transplantation: From Bench to Bedside. Int Rev Immunol. 2016;35(4):351-368. doi:10.3109/08830185.2016.1154051

9. Patel R, Terasaki PI. Significance of the positive crossmatch test in kidney transplantation. N Engl J Med. 1969;280(14):735-739. doi:10.1056/NEJM196904032801401
10. Cecka JM. The UNOS Scientific Renal Transplant Registry--ten years of kidney transplants. Clin Transpl. 1997:1-14.
11. Le Bas Bernardet S, Hourmant M, Valentin N, et al. Identification of the antibodies involved in B-cell crossmatch positivity in renal transplantation. Transplantation. 2003;75(4):477-482. doi:10.1097/01.TP.0000047311.77702.59
12. Rocha Y, Jaramillo A, Neumann J, Hacke K, Palou E, Torres J. Crossmatch assays in transplantation: Physical or virtual?: A review. Medicine (Baltimore). 2023;102(50):e36527. doi:10.1097/MD.0000000000036527
13. Ozdemir FN, Sezer S, Akcay A, et al. Panel reactive antibody positivity and associated HLA antibodies in Turkish renal transplant candidates. Transpl Immunol. 2004;12(2):185-188. doi:10.1016/j.trim.2003.11.003
14. Tait BD, Süsal C, Gebel HM, Nickerson PW, Zachary AA, Claas FHJ, et al. Consensus guidelines on the testing and clinical management issues associated with HLA and non-HLA antibodies in transplantation. Transplantation. 2013;95(1):19-47. doi:10.1097/TP.0b013e31827a19cc
15. Kashyap S, Tiwari AK, Chauhan R, Mehra S, Bhardwaj G, Rani N, et al. Pre-transplant compatibility testing: transition from complement-dependent cytotoxicity crossmatch to flow cytometry. Transpl Immunol. 2025;93:102281.
16. Tafulo S, Osório E, Mendes C, Liwski R. Complement-dependent cytotoxicity crossmatch in solid organ transplantation: The gold standard or golden history? Hum Immunol. 2024;85:110734. doi:10.1016/j.humimm.2023.110734
17. Cossarizza A, Chang HD, Radbruch A, et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition). Eur J Immunol. 2021;51(12):2708-3145. doi:10.1002/eji.202170126
18. Işıtmangil G, Kara M, Özel L, et al. Böbrek nakli öncesi lenfosit cross-match test sonuçlarının retrospektif değerlendirilmesi: mikrolenfositotoksisite yöntemi. Haydarpaşa Numune Eğitim ve Araştırma Hastanesi Tıp Dergisi. 2008;48(3):139-141.
19. Shyti E, Idrizi A, Sulcebe G. Histocompatibility testing for organ transplantation purposes in Albania: a single center experience. Balkan Med J. 2014;31(2):121-125. doi:10.5152/balkanmedj.2014.13045
20. Güngör TH. Kadavra vericili böbrek nakli için çağırılan hastalara yapılan farklı cross-match testlerinin karşılaştırılması. Yüksek Lisans Tezi. İzmir Katip Çelebi Üniversitesi, Sağlık Bilimleri Enstitüsü, Tıbbi Biyoloji ve Genetik Anabilim Dalı, 2017.
21. Garovoy MR, Rheinschmidt MA, Bigos M, Perkins H, Colombe B, Feduska NJ. Flow cytometry analysis: a high-sensitivity cross-match technique facilitating transplantation. Transplant Proc. 1983;15(3):1939-1944.
22. Worthington JE, Martin S, Al-Husseini DM, Dyer PA, Johnson RW. Posttransplantation production of donor HLA-specific antibodies as a predictor of renal transplant outcome. Transplantation. 2003;75(7):1034-1040. doi:10.1097/01.TP.0000055833.65192.3B
23. Lefaucheur C, Loupy A, Hill GS, et al. Preexisting donor-specific HLA antibodies predict outcome in kidney transplantation. J Am Soc Nephrol. 2010;21(8):1398-1406. doi:10.1681/ASN.2009101065
24. Loupy A, Lefaucheur C. Antibody-Mediated Rejection of Solid-Organ Allografts. N Engl J Med. 2018;379(12):1150-1160. doi:10.1056/NEJMra1802677
25. Abbas K, Mubarak M, Musharraf W, Hafeez AR, Aziz T, Zafar MN. Impact of low-level pretransplant donor-specific antibodies detected by the Luminex platform on acute rejection and long-term graft survival. World J Transplant. 2025;15(3):104308. doi:10.5500/wjt.v15.i3.104308
26. Ameur RF, Berkani LM, Belaid B, Habchi K, Saidani M, Djidjik R. Luminex Crossmatch in kidney transplantation. Scand J Immunol. 2023;98(1):e13279. doi:10.1111/sji.13279.
27. Tait BD. Detection of HLA Antibodies in Organ Transplant Recipients - Triumphs and Challenges of the Solid Phase Bead Assay. Front Immunol. 2016;7:570. doi:10.3389/fimmu.2016.00570
28. Tambur AR, Wiebe C. HLA antibody strength and its impact on organ transplant outcomes. Transplantation. 2022;106(6):1124-1135.
29. Altindal M, Guldan M, Ozbek L, et al. Desensitization in HLA-incompatible kidney transplant recipients: current strategies and emerging perspectives. Clin Kidney J. 2025;18(8):sfaf219. doi:10.1093/ckj/sfaf219