Die durchflusszytometrische Verträglichkeitsprobe – sensitivere Diagnostik zur Prädiktion von Transplantatabstoßungen

Zusammenfassung

Bei der Lebendnierenspende hat man die Chance, das immunologische Risiko vor Transplantation sehr detailliert zu testen. Neben der Bestimmung der humanen Leukozytenantigene (HLA) ist die Suche nach HLA-Antikörpern und die Verträglichkeitsprobe (Crossmatch) zwingend erforderlich, wobei in Deutschland derzeit als Standard die komplementvermittelte Zytotoxizität gemessen wird (CDC-Crossmatch). Es geht darum, herauszufinden, ob der Empfänger präformierte spenderspezifische Antikörper hat, die eine Transplantatabstoßung auslösen können. Im Ausland, beispielsweise in Großbritannien, ist dagegen das durchflusszytometrische Crossmatch inzwischen als Standardverfahren etabliert. Dieses wurde bereits 1983 beschrieben und ist sensitiver als das CDC-Crossmatch. Es gibt inzwischen umfangreiche Studien, die zeigen, dass ein positives durchflusszytometrisches Crossmatch ein erhöhtes Risiko für antikörpervermittelte Abstoßungen anzeigt. Dies konnte in unserer Kohorte bestätigt werden, in der ein positives durchflusszytometrisches Crossmatch für die T-Zellen ein 10,1-fach und für die B-Zellen ein 3,2-fach erhöhtes Risiko anzeigt.

Abstract

Prior to living kidney donation the immunological risk can be assessed very detailed. Apart from testing for human leukocyte antigens (HLA) the detection of HLA antibodies and the crossmatch are mandatory. In Germany, the standard method is still based on complement dependent cytotoxicity (CDC crossmatch). It is used to detect preformed donor-specific antibodies that could lead to transplant rejection. Abroad, for example in Great Britain, the flow cytometric crossmatch is currently established as a standard assay. This crossmatch was already described in 1983 and it is more sensitive than the CDC crossmatch. There are comprehensive data showing that a positive flow cytometric crossmatch indicates an increased risk of antibody mediated rejection. We could confirm these data in our own cohort; a positive T cell flow crossmatch indicated a 10.1-fold and a positive B cell crossmatch a 3.2-fold increased risk.

Schlüsselwörter

durchflusszytometrisches Crossmatch - Crossmatch zur Detektion von komplementvermittelter Zytotoxizität - antikörpervermittelte Abstoßung - Lebendnierenspende

Key words

flow cytometric crossmatch - CDC crossmatch - antibody mediated rejection - living kidney donation

• Literatur

• 1 Kissmeyer-Nielsen F, Olsen S, Petersen VP. et al. Hyperacute rejection of kidney allografts, associated with pre-existing humoral antibodies against donor cells. Lancet 1966; 2: 662-665
• 2 Morales-Buenrostro LE, Terasaki PI, Marino-Vazquez LA. et al. “Natural” human leukocyte antigen antibodies found in nonalloimmunized healthy males. Transplantation 2008; 86: 1111-1115
• 3 Zorn E, See SB. Polyreactive natural antibodies in transplantation. Curr Opin Organ Transplant 2017; 22: 8-13
• 4 Schnaidt M, Weinstock C, Jurisic M. et al. HLA antibody specification using single-antigen beads – a technical solution for the prozone effect. Transplantation 2011; 92: 510-515
• 5 Terasaki PI, McClelland JD. Microdroplet assay of human serum cytotoxins. Nature 1964; 204: 998-1000
• 6 Patel R, Terasaki PI. Significance of the positive crossmatch test in kidney transplantation. N Engl J Med 1969; 280: 735-739
• 7 Zachary AA, Klingman L, Thorne N. et al. Variations of the lymphocytotoxicity test. An evaluation of sensitivity and specificity. Transplantation 1995; 60: 498-503
• 8 Chapman JR, Taylor CJ, Ting A. et al. Immunoglobulin class and specificity of antibodies causing positive T cell crossmatches. Relationship to renal transplant outcome. Transplantation 1986; 42: 608-613
• 9 Nelken D, Cohen I, Furcaig I. A method to increase the sensitivity of the lymphocyte microcytotoxicity test. Transplantation 1970; 10: 346-347
• 10 Garovoy MR, Rheinschmidt MA, Bigos M. et al. Flow cytometry analysis: a high technology crossmatch technique facilitating transplantation. Transplant Proc 1983; 15: 1939-1944
• 11 Gloor JM, DeGoey S, Ploeger N. et al. Persistence of low levels of alloantibody after desensitization in crossmatch-positive living-donor kidney transplantation. Transplantation 2004; 78: 221-227
• 12 Rebibou JM, Bittencourt MC, Saint-Hillier Y. et al. T-cell flow-cytometry crossmatch and long-term renal graft survival. Clin Transplant 2004; 18: 558-563
• 13 Karpinski M, Rush D, Jeffery J. et al. Flow cytometric crossmatching in primary renal transplant recipients with a negative anti-human globulin enhanced cytotoxicity crossmatch. J Am Soc Nephrol 2001; 12: 2807-2814
• 14 Chapman JR, Deierhoi MH, Carter NP. et al. Analysis of flow cytometry and cytotoxicity crossmatches in renal transplantation. Transplant Proc 1985; 17: 2480-2481
• 15 Lindemann M, Nyadu B, Heinemann FM. et al. High negative predictive value of an amplified flow cytometry crossmatch before living donor kidney transplantation. Hum Immunol 2010; 71: 771-776
• 16 Heo WB, Kim CD, Won DI. HLA Class II-specific antibodies can react with T cells in flow cytometry crossmatch: a case report. Transplant Proc 2007; 39: 3485-3487
• 17 Cho YW, Cecka JM. Crossmatch tests – an analysis of UNOS data from 1991–2000. Clin Transpl 2001; 237-246
• 18 Tait BD, Susal C, Gebel HM. et al. Consensus guidelines on the testing and clinical management issues associated with HLA and non-HLA antibodies in transplantation. Transplantation 2013; 95: 19-47
• 19 Couzi L, Araujo C, Guidicelli G. et al. Interpretation of positive flow cytometric crossmatch in the era of the single-antigen bead assay. Transplantation 2011; 91: 527-535
• 20 Scornik JC, Clapp W, Patton PR. et al. Outcome of kidney transplants in patients known to be flow cytometry crossmatch positive. Transplantation 2001; 71: 1098-1102
• 21 Ishida H, Tanabe K, Furusawa M. et al. Evaluation of flow cytometric panel reactive antibody in renal transplant recipients – examination of 238 cases of renal transplantation. Transplant International 2005; 18: 163-168
• 22 Kerman RH, Susskind B, Buyse I. et al. Flow cytometry-detected IgG is not a contraindication to renal transplantation: IgM may be beneficial to outcome. Transplantation 1999; 68: 1855-1858
• 23 Wen R, Wu V, Dmitrienko S. et al. Biomarkers in transplantation: prospective, blinded measurement of predictive value for the flow cytometry crossmatch after negative antiglobulin crossmatch in kidney transplantation. Kidney Int 2006; 70: 1474-1481
• 24 Matinlauri IH, Kyllonen LE, Eklund BH. et al. Weak humoral posttransplant alloresponse after a well-HLA-matched cadaveric kidney transplantation. Transplantation 2004; 78: 198-204
• 25 Panigrahi A, Deka R, Bhowmik D. et al. Functional assessment of immune markers of graft rejection: a comprehensive study in live-related donor renal transplantation. Clin Transplant 2006; 20: 85-90
• 26 Takakura K, Kiuchi T, Kasahara M. et al. Clinical implications of flow cytometry crossmatch with T or B cells in living donor liver transplantation. Clin Transplant 2001; 15: 309-316
• 27 Limaye S, OʼKelly P, Harmon G. et al. Improved graft survival in highly sensitized patients undergoing renal transplantation after the introduction of a clinically validated flow cytometry crossmatch. Transplantation 2009; 87: 1052-1056
• 28 Lindemann M, Lenz V, Nyadu B. et al. Effect of AB0 incompatibility on T-cell flow cytometry cross-match results prior to living donor kidney transplantation. Cytometry B Clin Cytom 2016;
• 29 Lobo PI, Spencer CE, Stevenson WC. et al. The use of pronase-digested human leukocytes to improve specificity of the flow cytometric crossmatch. Transpl Int 1995; 8: 472-480
• 30 Forster O, Boltz-Nitulescu G. Digestion and resynthesis of receptors binding IgG-sensitized erythrocytes on rat macrophages. Immunology 1982; 47: 107-114
• 31 Bearden CM, Agarwal A, Book BK. et al. Pronase treatment facilitates alloantibody flow cytometric and cytotoxic crossmatching in the presence of rituximab. Hum Immunol 2004; 65: 803-809
• 32 Hetrick SJ, Schillinger KP, Zachary AA. et al. Impact of pronase on flow cytometric crossmatch outcome. Hum Immunol 2011; 72: 330-336
• 33 Brody JI, Beizer LH. Alteration of blood group antigens in leukemic lymphocytes. J Clin Invest 1965; 44: 1582-1589