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Thoracic Organ Transplantation: Laboratory Methods

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Transplantation Immunology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1034))

Abstract

Although great progress has been achieved in thoracic organ transplantation through the development of effective immunosuppression, there is still significant risk of rejection during the early post-transplant period, creating a need for routine monitoring for both acute antibody and cellular mediated rejection. The currently available multiplexed, microbead assays utilizing solubilized HLA antigens afford the capability of sensitive detection and identification of HLA and non-HLA specific antibodies. These assays are being used to assess the relative strength of donor specific antibodies; to permit performance of virtual crossmatches which can reduce the waiting time to transplantation; to monitor antibody levels during desensitization; and for heart transplants to monitor antibodies post-transplant. For cell mediated immune responses, the recent development of gene expression profiling has allowed noninvasive monitoring of heart transplant recipients yielding predictive values for acute cellular rejection. T cell immune monitoring in heart and lung transplant recipients has allowed individual tailoring of immunosuppression, particularly to minimize risk of infection. While the current antibody and cellular laboratory techniques have enhanced the ability to manage thoracic organ transplant recipients, future developments from improved understanding of microchimerism and graft tolerance may allow more refined allograft monitoring techniques.

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References

  1. Billingham RE, Brent L, Medawar PB (1953) Actively acquired tolerance of foreign cells. Nature 172(4379):603–606

    Article  PubMed  CAS  Google Scholar 

  2. Kobashigawa J, Mehra M, West L, Kerman R, George J, Rose M et al (2009) Report from a consensus conference on the sensitized patient awaiting heart transplantation. J Heart Lung Transplant 28(3):213–225. doi:S1053-2498(08)01785-3 [pii] 10.1016/j.healun.2008.12.017

    Article  PubMed  Google Scholar 

  3. Oyer PE, Stinson EB, Jamieson SW (1983) Cyclosporine in cardiac transplantation: a 21/2 year follow-up. Transplant Proc 15(Suppl 1):2546

    Google Scholar 

  4. Armitage JM, Kormos RL, Griffith BP, Hardesty RL, Fricker FJ, Stuart RS et al (1991) A clinical trial of FK 506 as primary and rescue immunosuppression in cardiac transplantation. Transplant Proc 23(1 Pt 2):1149–1152

    PubMed  CAS  Google Scholar 

  5. Cheung A, Menkis AH (1998) Cyclosporine heart transplantation. Transplant Proc 30(5):1881–1884

    Article  PubMed  CAS  Google Scholar 

  6. Stehlik J, Edwards LB, Kucheryavaya AY, Benden C, Christie JD, Dobbels F et al (2011) The Registry of the International Society for Heart and Lung Transplantation: twenty-­eighth Adult Heart Transplant Report—2011. J Heart Lung Transplant 30(10):1078–1094. doi:S1053-2498(11)01086-2 [pii] 10.1016/j.healun.2011.08.003

    Article  PubMed  Google Scholar 

  7. Michaels PJ, Espejo ML, Kobashigawa J, Alejos JC, Burch C, Takemoto S et al (2003) Humoral rejection in cardiac transplantation: risk factors, hemodynamic consequences and relationship to transplant coronary artery disease. J Heart Lung Transplant 22(1):58–69

    Article  PubMed  Google Scholar 

  8. Itescu S, Tung TC, Burke EM, Weinberg AD, Mancini D, Michler RE et al (1998) An immunological algorithm to predict risk of high-­grade rejection in cardiac transplant recipients. Lancet 352(9124):263–270

    Article  PubMed  CAS  Google Scholar 

  9. Nwakanma LU, Williams JA, Weiss ES, Russell SD, Baumgartner WA, Conte JV (2007) Influence of pretransplant panel-reactive antibody on outcomes in 8,160 heart transplant recipients in recent era. Ann Thorac Surg 84(5):1556–1562. doi:S0003-­4975(07)01217-9 [pii] 10.1016/j.athoracsur.2007.05.095; discussion 1562–1553

    Article  PubMed  Google Scholar 

  10. Tinckam KJ, Chandraker A (2006) Mechanisms and role of HLA and non-HLA alloantibodies. Clin J Am Soc Nephrol 1(3):404–414. doi:CJN.00270106 [pii] 10.2215/CJN.00270106

    Article  PubMed  CAS  Google Scholar 

  11. Schlaf G, Pollok-Kopp B, Manzke T, Schurat O, Altermann W (2010) Novel solid ­phase-­based ELISA assays contribute to an improved detection of anti-HLA antibodies and to an increased reliability of pre- and post-transplant crossmatching. NDT Plus 3(6):527–538

    Article  CAS  Google Scholar 

  12. Altermann WW, Seliger B, Sel S, Wendt D, Schlaf G (2006) Comparison of the established standard complement-dependent cytotoxicity and flow cytometric crossmatch assays with a novel ELISA-based HLA crossmatch procedure. Histol Histopathol 21(10):1115–1124

    PubMed  CAS  Google Scholar 

  13. Smith JD, Hamour IM, Banner NR, Rose ML (2007) C4d fixing, luminex binding antibodies—a new tool for prediction of graft failure after heart transplantation. Am J Transplant 7(12):2809–2815. doi:AJT1991 [pii] 10.1111/j.1600-6143.2007.01991.x

    Article  PubMed  CAS  Google Scholar 

  14. Won DI, Jung HD, Jung OJ, Huh S, Suh JS (2007) Flow cytometry PRA using lymphocyte pools from random donors. Cytometry B Clin Cytom 72(4):256–264. doi:10.1002/cyto.b.20175

    PubMed  Google Scholar 

  15. El-Awar N, Lee J, Terasaki PI (2005) HLA antibody identification with single antigen beads compared to conventional methods. Hum Immunol 66(9):989–997. doi:S0198-8859(05)00138-2 [pii] 10.1016/j.humimm.2005.07.005

    Article  PubMed  CAS  Google Scholar 

  16. Reinsmoen NL, Lai CH, Vo A, Cao K, Ong G, Naim M et al (2008) Acceptable donor-­specific antibody levels allowing for successful deceased and living donor kidney transplantation after desensitization therapy. Transplantation 86(6):820–825. doi:10.1097/TP.0b013e3181856f98 00007890-200809270-­00012 [pii]

    Article  PubMed  CAS  Google Scholar 

  17. Stehlik J, Islam N, Hurst D, Kfoury AG, Movsesian MA, Fuller A et al (2009) Utility of virtual crossmatch in sensitized patients awaiting heart transplantation. J Heart Lung Transplant 28(11):1129–1134

    Article  PubMed  Google Scholar 

  18. Yanagida R, Czer LS, Reinsmoen NL, Cao K, Rafiei M, De Robertis MA et al (2011) Impact of virtual cross match on waiting times for heart transplantation. Ann Thorac Surg 92(6):2104–2110. doiS0003-4975(11)01910-2 [pii] 10.1016/j.athoracsur.2011.07.082; discussion 2111

    Article  PubMed  Google Scholar 

  19. Zangwill S, Ellis T, Stendahl G, Zahn A, Berger S, Tweddell J (2007) Practical application of the virtual crossmatch. Pediatr Transplant 11(6):650–654. doi:PTR746 [pii] 10.1111/j.1399-3046.2007.00746.

    Article  PubMed  Google Scholar 

  20. Vaidya S, Hilson B, Sheldon S, Cano P, Fernandez-Vina M (2007) DP reactive antibody in a zero mismatch renal transplant pair. Hum Immunol 68(12):947–949. doi:S0198-8859(07)00465-X [pii] 10.1016/j.humimm.2007.10.013

    Article  PubMed  CAS  Google Scholar 

  21. Kobashigawa JA, Patel JK, Kittleson MM, Kawano MA, Kiyosaki KK, Davis SN et al (2010) The long-term outcome of treated sensitized patients who undergo heart transplantation. Clin Transplant 25(1):E61–E67. doi:10.1111/j.1399-0012.2010.01334.x

    Article  PubMed  Google Scholar 

  22. Patel J, Everly M, Chang D, Kittleson M, Reed E, Kobashigawa J (2011) Reduction of alloantibodies via proteosome inhibition in cardiac transplantation. J Heart Lung Transplant 30(12):1320–1326. doi:S1053-2498(11)01110-7 [pii] 10.1016/j.healun.2011.08.009

    Article  PubMed  Google Scholar 

  23. Kauke T, Kaczmarek I, Dick A, Schmoeckel M, Deutsch MA, Beiras-Fernandez A et al (2009) Anti-MICA antibodies are related to adverse outcome in heart transplant recipients. J Heart Lung Transplant 28(4):305–311. doi:S1053-2498(09)00024-2 [pii] 10.1016/j.healun.2009.01.003

    Article  PubMed  Google Scholar 

  24. Angaswamy N, Saini D, Ramachandran S, Nath DS, Phelan D, Hachem R et al (2010) Development of antibodies to human leukocyte antigen precedes development of antibodies to major histocompatibility class I-related chain A and are significantly associated with development of chronic rejection after human lung transplantation. Hum Immunol 71(6):560–565. doi:S0198-8859(10)00064-9 [pii] 10.1016/j.humimm.2010.02.021

    Article  PubMed  CAS  Google Scholar 

  25. Reinsmoen NL, Lai C-H, Mirocha J, Cao K, Ong GD, Naim M et al (2013) Increased negative impact of anti-angiotensin type-1 receptor antibodies together with de novo donor HLA specific antibodies on graft outcome in heart transplant recipients. J Heart Lung Transplant 32(4S):S113

    Google Scholar 

  26. Hiemann NE, Meyer R, Wellnhofer E, Schoenemann C, Heidecke H, Lachmann N, Hetzer R et al. (2012) Dragun Non-HLA antibodies targeting vascular receptors enhance alloimmune response and microvasculopathy after heart transplantation. Transplantation. Nov 15;94(9):919–24

    Google Scholar 

  27. Faulk WP, Rose M, Meroni PL, Del Papa N, Torry RJ, Labarrere CA et al (1999) Antibodies to endothelial cells identify myocardial damage and predict development of coronary artery disease in patients with transplanted hearts. Hum Immunol 60(9):826–832

    Article  PubMed  CAS  Google Scholar 

  28. Jurcevic S, Ainsworth ME, Pomerance A, Smith JD, Robinson DR, Dunn MJ et al (2001) Antivimentin antibodies are an independent predictor of transplant-associated coronary artery disease after cardiac transplantation. Transplantation 71(7):886–892

    Article  PubMed  CAS  Google Scholar 

  29. Goers TA, Ramachandran S, Aloush A, Trulock E, Patterson GA, Mohanakumar T (2008) De novo production of K-alpha1 tubulin-­specific antibodies: role in chronic lung allograft rejection. J Immunol 180(7):4487–4494. doi:180/7/4487 [pii]

    PubMed  CAS  Google Scholar 

  30. Iwata T, Philipovskiy A, Fisher AJ, Presson RG Jr, Chiyo M, Lee J et al (2008) Anti-type V collagen humoral immunity in lung transplant primary graft dysfunction. J Immunol 181(8):5738–5747. doi:181/8/5738 [pii]

    PubMed  CAS  Google Scholar 

  31. Zhang Q, Cecka JM, Gjertson DW, Ge P, Rose ML, Patel JK, Ardehali A et al (2011) HLA and MICA: targets of antibody-mediated rejection in heart transplantation. Transplantation 91(10):1153–1158. doi:10.1097/TP.0b013e3182157d60 00007890-201105270-00015 [pii]

    Article  PubMed  CAS  Google Scholar 

  32. Mahesh B, Leong HS, McCormack A, Sarathchandra P, Holder A, Rose ML (2007) Autoantibodies to vimentin cause accelerated rejection of cardiac allografts. Am J Pathol 170(4):1415–1427. doi:S0002-9440(10)61355-7 [pii] 10.2353/ajpath.2007.060728

    Article  PubMed  CAS  Google Scholar 

  33. Chin C, Chen G, Sequeria F, Berry G, Siehr S, Bernstein D et al (2010) Clinical usefulness of a novel C1q assay to detect immunoglobulin G antibodies capable of fixing complement in sensitized pediatric heart transplant patients. J Heart Lung Transplant 30(2):158–163. doi:S1053-2498(10)00508-5 [pii] 10.1016/j.healun.2010.08.020

    Article  PubMed  Google Scholar 

  34. Chen G, Sequeira F, Tyan DB (2011) Novel C1q assay reveals a clinically relevant subset of human leukocyte antigen antibodies independent of immunoglobulin G strength on single antigen beads. Hum Immunol 72(10):849–858. doi:S0198-8859(11)00171-6 [pii] 10.1016/j.humimm.2011.07.001

    Article  PubMed  CAS  Google Scholar 

  35. Kobashigawa J, Crespo-Leiro MG, Ensminger SM, Reichenspurner H, Angelini A, Berry G et al (2011) Report from a consensus conference on antibody-mediated rejection in heart transplantation. J Heart Lung Transplant 30(3):252–269. doi:S1053-2498(10)00731-­X [pii] 10.1016/j.healun.2010.11.003

    Article  PubMed  Google Scholar 

  36. Mehra MR, Parameshwar J (2010) Gene expression profiling and cardiac allograft rejection monitoring: is IMAGE just a mirage? J Heart Lung Transplant 29(6):599–602

    Article  PubMed  Google Scholar 

  37. McWilliams TJ, Williams TJ, Whitford HM, Snell GI (2008) Surveillance bronchoscopy in lung transplant recipients: risk versus benefit. J Heart Lung Transplant 27(11):1203–1209. doi:S1053-2498(08)00581-0 [pii] 10.1016/j.healun.2008.08.004

    Article  PubMed  Google Scholar 

  38. Deng MC, Eisen HJ, Mehra MR, Billingham M, Marboe CC, Berry G et al (2006) Noninvasive discrimination of rejection in cardiac allograft recipients using gene expression profiling. Am J Transplant 6(1):150–160

    Article  PubMed  CAS  Google Scholar 

  39. Pham MX, Teuteberg JJ, Kfoury AG, Starling RC, Deng MC, Cappola TP et al (2010) Gene-expression profiling for rejection surveillance after cardiac transplantation. N Engl J Med 362(20):1890–1900

    Article  PubMed  CAS  Google Scholar 

  40. Kowalski RJ, Post DR, Mannon RB, Sebastian A, Wright HI, Sigle G et al (2006) Assessing relative risks of infection and rejection: a meta-­analysis using an immune function assay. Transplantation 82(5):663–668. doi:10.1097/01.tp.0000234837.02126.70 00007890-200609150-00011 [pii]

    Article  PubMed  Google Scholar 

  41. Kobashigawa JA, Kiyosaki KK, Patel JK, Kittleson MM, Kubak BM, Davis SN et al (2010) Benefit of immune monitoring in heart transplant patients using ATP production in activated lymphocytes. J Heart Lung Transplant 29(5):504–508. doi:S1053-2498(09)01536-8 [pii] 10.1016/j.healun.2009.12.01

    Article  PubMed  Google Scholar 

  42. Rossano JW, Denfield SW, Kim JJ, Price JF, Jefferies JL, Decker JA et al (2009) Assessment of the Cylex ImmuKnow cell function assay in pediatric heart transplant patients. J Heart Lung Transplant 28(1):26–31. doi:S1053-2498(08)00734-1 [pii] 10.1016/j.healun.2008.10.001

    Article  PubMed  Google Scholar 

  43. Bhorade SM, Janata K, Vigneswaran WT, Alex CG, Garrity ER (2008) Cylex ImmuKnow assay levels are lower in lung transplant recipients with infection. J Heart Lung Transplant 27(9):990–994. doi:S1053-2498(08)00477-4 [pii] 10.1016/j.healun.2008.06.005

    Article  PubMed  Google Scholar 

  44. Husain S, Raza K, Pilewski JM, Zaldonis D, Crespo M, Toyoda Y et al (2009) Experience with immune monitoring in lung transplant recipients: correlation of low immune function with infection. Transplantation 87(12):1852–1857. doi:10.1097/TP.0b013e3181a75ad2 00007890-200906270-00015 [pii]

    Article  PubMed  CAS  Google Scholar 

  45. Gautam A, Morrissey PE, Brem AS, Fischer SA, Gohh RY, Yango AF et al (2006) Use of an immune function assay to monitor immunosuppression for treatment of post-transplant lymphoproliferative disorder. Pediatr Transplant 10(5):613–616. doi:PTR510 [pii] 10.1111/j.1399-3046.2006.00510.x

    Article  PubMed  Google Scholar 

  46. Masters RG, Davies RA, Veinot JP, Hendry PJ, Smith SJ, de Bold AJ (1999) Discoordinate modulation of natriuretic peptides during acute cardiac allograft rejection in humans. Circulation 100(3):287–291

    Article  PubMed  CAS  Google Scholar 

  47. O’Neill JO, McRae AT 3rd, Troughton RW, Ng K, Taylor DO, Yamani MH et al (2005) Brain natriuretic peptide levels do not correlate with acute cellular rejection in De Novo orthotopic heart transplant recipients. J Heart Lung Transplant 24(4):416–420. doi:S105324980400035X [pii] 10.1016/j.healun.2003.12.006

    Article  PubMed  Google Scholar 

  48. Kittleson MM, Skojec DV, Wittstein IS, Champion HC, Judge DP, Barouch LA et al (2009) The change in B-type natriuretic peptide levels over time predicts significant rejection in cardiac transplant recipients. J Heart Lung Transplant 28(7):704–709. doi:S1053-2498(09)00241-1 [pii] 10.1016/j.healun.2009.04.019

    Article  PubMed  Google Scholar 

  49. Claudius I, Lan YT, Chang RK, Wetzel GT, Alejos J (2003) Usefulness of B-type natriuretic peptide as a noninvasive screening tool for cardiac allograft pathology in pediatric heart transplant recipients. Am J Cardiol 92(11):1368–1370. doi:S0002914903012049 [pii]

    Article  PubMed  CAS  Google Scholar 

  50. Rossano JW, Denfield SW, Kim JJ, Price JF, Jefferies JL, Decker JA et al (2008) B-type natriuretic peptide is a sensitive screening test for acute rejection in pediatric heart transplant patients. J Heart Lung Transplant 27(6):649–654. doi:S1053-2498(08)00248-9 [pii] 10.1016/j.healun.2008.03.008

    Article  PubMed  Google Scholar 

  51. Mehra MR, Uber PA, Walther D, Vesely M, Wohlgemuth JG, Prentice J et al (2006) Gene expression profiles and B-type natriuretic peptide elevation in heart transplantation: more than a hemodynamic marker. Circulation 114(1 Suppl):I21–I26. doi:114/1_suppl/I-21 [pii] 10.1161/CIRCULATIONAHA.105.000513

    PubMed  Google Scholar 

  52. Martinez-Dolz L, Almenar L, Reganon E, Vila V, Sanchez-Soriano R, Martinez-Sales V et al (2009) What is the best biomarker for diagnosis of rejection in heart transplantation? Clin Transplant 23(5):672–680. doi:CTR1074 [pii] 10.1111/j.1399-0012.2009.01074.x

    Article  PubMed  CAS  Google Scholar 

  53. Balduini A, Campana C, Ceresa M, Arbustini E, Bosoni T, Serio A et al (2003) Utility of biochemical markers in the follow-up of heart transplant recipients. Transplant Proc 35(8):3075–3078. doi:S0041134503011394 [pii]

    Article  PubMed  CAS  Google Scholar 

  54. Labarrere CA, Lee JB, Nelson DR, Al-Hassani M, Miller SJ, Pitts DE (2002) C-reactive protein, arterial endothelial activation, and development of transplant coronary artery disease: a prospective study. Lancet 360(9344):1462–1467. doi:S0140-6736(02)11473-5 [pii] 10.1016/S0140-6736(02)11473-5

    Article  PubMed  CAS  Google Scholar 

  55. Raichlin ER, McConnell JP, Lerman A, Kremers WK, Edwards BS, Kushwaha SS et al (2007) Systemic inflammation and metabolic syndrome in cardiac allograft vasculopathy. J Heart Lung Transplant 26(8):826–833. doi:S1053-2498(07)00400-7 [pii] 10.1016/j.healun.2007.05.008

    Article  PubMed  Google Scholar 

  56. Hognestad A, Endresen K, Wergeland R, Stokke O, Geiran O, Holm T et al (2003) Plasma C-reactive protein as a marker of cardiac allograft vasculopathy in heart transplant recipients. J Am Coll Cardiol 42(3):477–482. doi:S0735109703006454 [pii]

    Article  PubMed  CAS  Google Scholar 

  57. Sahota A, Gao S, Hayes J, Jindal RM (2000) Microchimerism and rejection: a meta-­analysis. Clin Transplant 14(4 Pt 1):345–350

    Article  PubMed  CAS  Google Scholar 

  58. Quaini F, Urbanek K, Beltrami AP, Finato N, Beltrami CA, Nadal-Ginard B et al (2002) Chimerism of the transplanted heart. N Engl J Med 346(1):5–15. doi:10.1056/NEJMoa012081346/1/5 [pii]

    Article  PubMed  Google Scholar 

  59. Pujal JM, Gallardo D (2008) PCR-based methodology for molecular microchimerism detection and quantification. Exp Biol Med (Maywood) 233(9):1161–1170. doi:0802-RM-­35 [pii] 10.3181/0802-RM-35

    Article  CAS  Google Scholar 

  60. Schmidt-Lucke C, Aicher A, Romagnani P, Gareis B, Romagnani S, Zeiher AM et al (2007) Specific recruitment of CD4 + CD25++ regulatory T cells into the allograft in heart transplant recipients. Am J Physiol Heart Circ Physiol 292(5):H2425–H2431. doi:01197.2006 [pii] 10.1152/ajpheart.01197.2006

    Article  PubMed  CAS  Google Scholar 

  61. Chen M, Mohtize M, Mattei MF, Villemot JP, Kohler C, Faure GC et al (2010) Reduced levels of both circulating CD4+ CD25+ CD127(low/neg) and CD4+ CD8(neg) invariant natural killer regulatory T cells in stable heart transplant recipients. Clin Exp Immunol 163(1):104–112. doi:10.1111/j.1365-2249.2010.04281.x

    Article  PubMed  Google Scholar 

  62. http://optn.transplant.hrsa.gov/resources/allocationcalculators.asp?index=78 Accessed on 1st May 2013

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Cedars-Sinai Heart Institute, Los Angeles, CA, USA

    Jignesh K. Patel & Jon A. Kobashigawa

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  1. Jignesh K. Patel
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  1. , Immunogenetics Laboratory, Johns Hopkins University School of Medic, E. Monument Street 2041, Baltimore, 21205, Maryland, USA

    Andrea A. Zachary

  2. , Immunogenetics Laboratory, Johns Hopkins University School of Medic, E. Monument Street 2041, Baltimore, 21205, Maryland, USA

    Mary S. Leffell

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Patel, J.K., Kobashigawa, J.A. (2013). Thoracic Organ Transplantation: Laboratory Methods. In: Zachary, A., Leffell, M. (eds) Transplantation Immunology. Methods in Molecular Biology, vol 1034. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-493-7_7

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