Mass cytometry reveals a distinct immunoprofile of operational tolerance in pediatric liver transplantation
Audrey H. Lau
Division of Pediatric Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, Lucile Packard Children's Hospital, Stanford, CA, USA
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Search for more papers by this authorMatthew J. Vitalone
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Search for more papers by this authorKelly Haas
Division of Pediatric Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, Lucile Packard Children's Hospital, Stanford, CA, USA
Search for more papers by this authorTodd Shawler
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Search for more papers by this authorCarlos O. Esquivel
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Search for more papers by this authorWilliam E. Berquist
Division of Pediatric Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, Lucile Packard Children's Hospital, Stanford, CA, USA
Search for more papers by this authorOlivia M. Martinez
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Stanford Immunology, Stanford University School of Medicine, Stanford, CA, USA
Search for more papers by this authorRicardo O. Castillo
Division of Pediatric Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, Lucile Packard Children's Hospital, Stanford, CA, USA
Search for more papers by this authorCorresponding Author
Sheri M. Krams
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Stanford Immunology, Stanford University School of Medicine, Stanford, CA, USA
Correspondence
Sheri M. Krams, Transplant Immunobiology Lab, Stanford University School of Medicine, Stanford, CA, USA.
Email: [email protected]
Search for more papers by this authorAudrey H. Lau
Division of Pediatric Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, Lucile Packard Children's Hospital, Stanford, CA, USA
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Search for more papers by this authorMatthew J. Vitalone
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Search for more papers by this authorKelly Haas
Division of Pediatric Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, Lucile Packard Children's Hospital, Stanford, CA, USA
Search for more papers by this authorTodd Shawler
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Search for more papers by this authorCarlos O. Esquivel
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Search for more papers by this authorWilliam E. Berquist
Division of Pediatric Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, Lucile Packard Children's Hospital, Stanford, CA, USA
Search for more papers by this authorOlivia M. Martinez
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Stanford Immunology, Stanford University School of Medicine, Stanford, CA, USA
Search for more papers by this authorRicardo O. Castillo
Division of Pediatric Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, Lucile Packard Children's Hospital, Stanford, CA, USA
Search for more papers by this authorCorresponding Author
Sheri M. Krams
Division of Abdominal Transplantation, Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA
Stanford Immunology, Stanford University School of Medicine, Stanford, CA, USA
Correspondence
Sheri M. Krams, Transplant Immunobiology Lab, Stanford University School of Medicine, Stanford, CA, USA.
Email: [email protected]
Search for more papers by this authorAbstract
Long-term IS in transplant patients has significant morbidity, poorer quality of life, and substantial economic costs. TOL, defined as graft acceptance without functional impairment in the absence of IS, has been achieved in some pediatric LT recipients. Using mass cytometry, peripheral blood immunotyping was performed to characterize differences between tolerant patients and patients who are stable on single-agent IS. Single-cell mass cytometry was performed using blood samples from a single-center pediatric LT population of operationally tolerant patients to comprehensively characterize the immune cell populations in the tolerant state compared with patients on chronic low-dose IS. Specific T-cell populations of interest were confirmed by flow cytometry. This high-dimensional phenotypic analysis revealed distinct immunoprofiles between transplant populations as well as a CD4+ TOT (CD4+CD5+CD25+CD38−/loCD45RA) that correlates with tolerance in pediatric LT recipients. In TOL patients, the TOT was significantly increased as compared to patients stable on low levels of IS. This TOT cell was confirmed by flow cytometry and is distinct from classic Treg cells. These results demonstrate the power of mass cytometry to discover significant immune cell signatures that have diagnostic potential.
Supporting Information
| Filename | Description |
|---|---|
| petr12795-sup-0001-FigS1.tifTIFF image, 5.8 MB | |
| petr12795-sup-0002-FigS2.tiffTIFF image, 321.9 KB | |
| petr12795-sup-0003-FigS3.tiffTIFF image, 567.9 KB | |
| petr12795-sup-0004-FigS4.tiffTIFF image, 273.7 KB | |
| petr12795-sup-0005-TableS1.docxWord document, 21.1 KB | |
| petr12795-sup-0006-TableS2.docxWord document, 24.8 KB | |
| petr12795-sup-0007-TableS3.docxWord document, 29.2 KB | |
| petr12795-sup-0008-Legends.docxWord document, 19.9 KB |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
References
- 1Asfar S, Metrakos P, Fryer J, et al. An analysis of late deaths after liver transplantation. Transplantation. 1996; 61: 1377–1381.
- 2Gelson W, Hoare M, Dawwas MF, Vowler S, Gibbs P, Alexander G. The pattern of late mortality in liver transplant recipients in the United Kingdom. Transplantation. 2011; 91: 1240–1244.
- 3Schonder KS, Mazariegos GV, Weber RJ. Adverse effects of immunosuppression in pediatric solid organ transplantation. Pediatr Drugs. 2010; 12: 35–49.
- 4Rasmussen A, Davies HF, Jamieson NV, Evans DB, Calne RY. Combined transplantation of liver and kidney from the same donor protects the kidney from rejection and improves kidney graft survival. Transplantation. 1995; 59: 919–921.
- 5Katznelson S, Cecka JM. The liver neither protects the kidney from rejection nor improves kidney graft survival after combined liver and kidney transplantation from the same donor. Transplantation. 1996; 61: 1403–1405.
- 6Saidman SL, Duquesnoy RJ, Demetris AJ, et al. Combined liver-kidney transplantation and the effect of preformed lymphocytotoxic antibodies. Transpl Immunol. 1994; 2: 61–67.
- 7Benitez C, Londono MC, Miquel R, et al. Prospective multicenter clinical trial of immunosuppressive drug withdrawal in stable adult liver transplant recipients. Hepatology. 2013; 58: 1824–1835.
- 8Feng S, Ekong UD, Lobritto SJ, et al. Complete immunosuppression withdrawal and subsequent allograft function among pediatric recipients of parental living donor liver transplants. JAMA. 2012; 307: 283–293.
- 9Mazariegos GV, Reyes J, Marino IR, et al. Weaning of immunosuppression in liver transplant recipients. Transplantation. 1997; 63: 243–249.
- 10Braza F, Dugast E, Panov I, et al. Central role of CD45RA- Foxp3hi memory regulatory T cells in clinical kidney transplantation tolerance. J Am Soc Nephrol. 2015; 26: 1795–1805.
- 11Demetris AJ, Lunz JG III, Randhawa P, Wu T, Nalesnik M, Thomson AW. Monitoring of human liver and kidney allograft tolerance: a tissue/histopathology perspective. Transpl Int. 2009; 22: 120–141.
- 12Germani G, Rodriguez-Castro K, Russo FP, et al. Markers of acute rejection and graft acceptance in liver transplantation. World J Gastroenterol. 2015; 21: 1061–1068.
- 13Li L, Wozniak LJ, Rodder S, et al. A common peripheral blood gene set for diagnosis of operational tolerance in pediatric and adult liver transplantation. Am J Transplant. 2012; 12: 1218–1228.
- 14Li Y, Koshiba T, Yoshizawa A, et al. Analyses of peripheral blood mononuclear cells in operational tolerance after pediatric living donor liver transplantation. Am J Transplant. 2004; 4: 2118–2125.
- 15Martinez-Llordella M, Lozano JJ, Puig-Pey I, et al. Using transcriptional profiling to develop a diagnostic test of operational tolerance in liver transplant recipients. J Clin Invest. 2008; 118: 2845–2857.
- 16Martinez-Llordella M, Puig-Pey I, Orlando G, et al. Multiparameter immune profiling of operational tolerance in liver transplantation. Am J Transplant. 2007; 7: 309–319.
- 17Sagoo P, Perucha E, Sawitzki B, et al. Development of a cross-platform biomarker signature to detect renal transplant tolerance in humans. J Clin Invest. 2010; 120: 1848–1861.
- 18Schulz-Juergensen S, Marischen L, Wesch D, et al. Markers of operational immune tolerance after pediatric liver transplantation in patients under immunosuppression. Pediatr Transplant. 2013; 17: 348–354.
- 19Tokita D, Mazariegos GV, Zahorchak AF, et al. High PD-L1/CD86 ratio on plasmacytoid dendritic cells correlates with elevated T-regulatory cells in liver transplant tolerance. Transplantation. 2008; 85: 369–377.
- 20Stenard F, Nguyen C, Cox K, et al. Decreases in circulating CD4+CD25hiFOXP3+ cells and increases in intragraft FOXP3+ cells accompany allograft rejection in pediatric liver allograft recipients. Pediatr Transplant. 2009; 13: 70–80.
- 21Bendall SC, Simonds EF, Qiu P, et al. Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum. Science. 2011; 332: 687–696.
- 22Bruggner RV, Bodenmiller B, Dill DL, Tibshirani RJ, Nolan GP. Automated identification of stratifying signatures in cellular subpopulations. Proc Natl Acad Sci USA. 2014; 111: E2770–E2777.
- 23 R Core Team (2014). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL http://www.R-project.org/.
- 24Tibshirani R, Hastie T, Narasimhan B, Chu G. Diagnosis of multiple cancer types by shrunken centroids of gene expression. Proc Natl Acad Sci USA. 2002; 99: 6567–6572.
- 25Tibshirani R, Society RS. Regression and shrinkage via the Lasso. J R Stat Soc Series B. 1996: 58: 267–288.
- 26Becker RA, Chambers JM, Wilks AR. The New S Language. USA: Wadsworth and Brooks/Cole Advanced Books & Software; 1988.
- 27Mardia KV, Kent JT, Bibby JM. Multivariate analysis. Analysis. 1979; 97: 1–4.
- 28Venables WN, Ripley BD. Modern Applied Statistics with S. Vol 53. 4th edn. New York: Springer-Verlag; 2002, 489p.
- 29Lau AH, Abe M, Thomson AW. Ethanol affects the generation, cosignaling molecule expression, and function of plasmacytoid and myeloid dendritic cell subsets in vitro and in vivo. J Leukoc Biol. 2006; 79: 941–953.
- 30Lau AH, Thomson AW, Colvin BL. Chronic ethanol exposure affects in vivo migration of hepatic dendritic cells to secondary lymphoid tissue. Hum Immunol. 2007; 68: 577–585.
- 31Pham B, Piard-Ruster K, Silva R, et al. Changes in natural killer cell subsets in pediatric liver transplant recipients. Pediatr Transplant. 2012; 16: 176–182.
- 32Lerut J, Sanchez-Fueyo A. An appraisal of tolerance in liver transplantation. Am J Transplant. 2006; 6: 1774–1780.
- 33Puig-Pey I, Bohne F, Benítez C, et al. Characterization of γδ T cell subsets in organ transplantation. Transpl Int. 2010; 23: 1045–1055.
- 34Pankratz S, Bittner S, Herrmann AM, et al. Human CD4+HLA-G+ regulatory T cells are potent suppressors of graft-versus-host disease in vivo. FASEB J. 2014; 28: 3435–3445.
- 35Gagliani N, Magnani CF, Huber S, et al. Coexpression of CD49b and LAG-3 identifies human and mouse T regulatory type 1 cells. Nat Med. 2013; 19: 739–746.
- 36Henderson Jacob G, Opejin A, Jones A, Gross C, Hawiger D. CD5 Instructs extrathymic regulatory T cell development in response to self and tolerizing antigens. Immunity. 2015; 42: 471–483.
- 37Yoshitomi M, Koshiba T, Haga H, et al. Requirement of protocol biopsy before and after complete cessation of immunosuppression after liver transplantation. Transplantation. 2009; 87: 606–614.
Citing Literature
