1 / 62

PRA = 36% (21/58)

1. 8. 1. 1. 1. 1. 1. 1. 1. 1. 1. 8. 8. 1. 1. 1. 1. 1. 8. 1. 1. 1. 1. 1. 1. 8. 1. 8. 1. 1. 1. 1. 1. 1. 8. 1. 1. 8. 1. 1. 1. 1. 1. 1. 8. 8. 1. 1. 1. 8. 1. 1. 1. 1. 1. 1. 1. 1. 1. 8. 8. 1. 1. 1. 1. 8. 1. 1. 1. 1. 1. 1. 1. 1. 1.

ghoyle
Download Presentation

PRA = 36% (21/58)

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. 1 8 1 1 1 1 1 1 1 1 1 8 8 1 1 1 1 1 8 1 1 1 1 1 1 8 1 8 1 1 1 1 1 1 8 1 1 8 1 1 1 1 1 1 8 8 1 1 1 8 1 1 1 1 1 1 1 1 1 8 8 1 1 1 1 8 1 1 1 1 1 1 1 1 1 1 1 1 8 8 1 1 1 8 1 8 1 1 1 1 1 8 1 1 1 1 1 8 1 1 8 8 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 PRA = 36% (21/58) Anti-A11 and B44

  2. 1 8 1 1 1 1 1 1 1 1 1 8 8 1 1 1 1 1 8 1 1 1 1 1 1 8 1 8 1 1 1 1 1 1 8 1 1 8 1 1 1 1 1 1 8 8 1 1 1 8 1 1 1 1 1 1 1 1 1 8 8 1 1 1 1 8 1 1 1 1 1 1 1 1 1 1 1 1 8 8 1 1 1 8 1 8 1 1 1 1 1 8 1 1 1 1 1 8 1 1 8 8 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 PRA = 36% (21/58) Anti-A11 and B44

  3. 8 8 1 8 1 1 8 1 8 1 1 8 8 8 1 8 1 1 8 1 1 8 1 1 8 8 1 8 8 1 1 8 8 1 8 1 1 8 1 1 1 1 1 1 8 8 8 1 8 8 1 1 8 8 8 1 1 1 1 8 8 1 8 8 1 8 1 1 1 8 1 1 1 8 1 8 1 1 8 8 8 8 1 8 8 8 1 1 1 1 1 8 8 1 8 8 1 8 1 1 8 8 1 1 8 8 8 1 1 8 1 1 8 1 1 1 8 1 8 1 PRA = 95% (55/58) Specificity?

  4. Flow Cytometry Crossmatch B cell T cell

  5. Flow Cytometry Crossmatch FITC-a-IgG B cell T cell

  6. Flow Cytometry Crossmatch FITC-a-IgG FITC-a-IgG B cell T cell

  7. Flow Cytometry Crossmatch Anti-CD3 Anti-CD19 FITC-a-IgG FITC-a-IgG B cell T cell

  8. Flow Cytometry Crossmatch FITC-a-IgG FITC-a-IgG B cell Anti-CD3 T cell Anti-CD19 Detect fluorescent labels by flow cytometry

  9. Flow Crossmatch Implemented in Halifax, June 2010

  10. Flow Cytometry Crossmatch T cell X-match B cell X-match Negative Gating strategy Weak positive Strong positive FITC-a-IgG FITC-a-IgG

  11. Karpinski et al. JASN 2001 • Retrospective flow cytometry crossmatch study • 249 patients transplanted (June 1992 and June 2000) with negative CDC-AHG crossmatch

  12. Karpinski et al. JASN 2001

  13. Strategies used to avoid/minimize transplant rejection • HLA typing and matching of recipient/donor pairs • Detection of donor specific HLA antibodies. • Lymphocyte crossmatch • Complement dependent cytotoxicity (CDC) crossmatch. • Flow cytometrycrossmatch (newer technique, much more sensitive) • Virtual crossmatch • Identification of HLA antibodies in recipient serum by solid phase assay • HLA typing of the donor (and recipient) • Correlation of recipient HLA antibodies and donor/recipient typing

  14. HLA antibody identification by Luminex (solid phase) Assay HLA antigen coated microspheres Tells the instrument which bead is being examined 2 lasers Tells the instrument how much antibody is bound to the bead

  15. HLA antibody detection by Luminex assay 1 2 3 4 5 6 7 8 9 10

  16. HLA antibody detection by Luminex assay 1 2 3 4 5 6 7 8 9 10 A1 A2 A3 A11 A23 A24 A25 A26 A29 A30

  17. HLA antibody detection by Luminex assay 8 9 7 A26 A29 A25 4 10 10 6 8 A11 9 1 5 A30 A30 A24 A26 1 A29 A1 A23 2 A1 5 3 6 A2 7 A23 A3 A24 3 4 2 A25 A3 A11 A2

  18. HLA antibody detection by Luminex assay 8 9 7 A26 A29 A25 4 10 10 6 8 A11 9 1 5 A30 A30 A24 A26 1 A29 A1 A23 2 A1 5 3 6 A2 7 A23 A3 A24 3 4 2 A25 A3 A11 A2

  19. HLA antibody detection by Luminex assay 8 9 7 A26 A29 A25 4 10 10 6 8 A11 9 1 5 A30 A30 A24 A26 1 A29 A1 A23 2 A1 5 3 6 A2 7 A23 A3 A24 3 4 2 A25 A3 A11 A2

  20. HLA antibody detection by Luminex assay 8 9 7 A26 A29 A25 4 10 10 6 8 A11 9 1 5 A30 A30 A24 A26 1 A29 A1 A23 2 A1 5 3 6 A2 7 A23 A3 A24 3 4 2 A25 A3 A11 A2

  21. HLA antibody detection by Luminex assay PE-a-IgG 8 9 7 A26 A29 A25 4 10 10 6 8 A11 9 1 5 A30 A30 A24 A26 1 A29 A1 A23 2 A1 5 3 6 A2 7 A23 A3 A24 3 4 2 A25 A3 A11 A2

  22. HLA antibody detection by Luminex assay 8 9 7 A26 A29 A25 4 10 10 6 8 A11 9 1 5 A30 A30 A24 A26 1 A29 A1 A23 2 A1 5 3 6 A2 7 A23 A3 A24 3 4 2 A25 A3 A11 A2

  23. Patient Case

  24. HLA Class I antibody analysis Patient A3,31 B7,60 DR1,14 (52) DQB5,6

  25. HLA Class I antibody analysis Patient A3,31 B7,60 DR1,14 (52) DQB5,6 Donor A1, B8 DR7,17 (53,52) DQB2

  26. HLA Class I antibody analysis Patient A3,31 B7,60 DR1,14 (52) DQB5,6 Donor A1, B8 DR7,17 (53,52) DQB2 Unacceptable antigens: A1, A36, B8

  27. HLA Class II antibody analysis Patient A3,31 B7,60 DR1,14 (52) DQB5,6 Donor A1, B8 DR7,17 (53,52) DQB2

  28. HLA Class II antibody analysis Patient A3,31 B7,60 DR1,14 (52) DQB5,6 Donor A1, B8 DR7,17 (53,52) DQB2 Unacceptable antigens: DR7, DR53, DQ2

  29. What is the clinical relevance of donor specific HLA antibodies detected pre-transplant by solid phase assay?

  30. Amico et al. Transplantation 2009 Significant increase in biopsy proven AMR in patients with pre-transplant DSA

  31. Lefaucheur et al. JASN 2010 Significant decrease in graft survival in patients with pre-transplant DSA Class I and Class II DSA confer similar risk.

  32. What about PRA?(probability of a positive crossmatch)

  33. Calculated PRA • calculated PRA (cPRA) is based on the unacceptable HLA antigens listed for a patient • cPRA is determined using an established algorithm (Zachary et al) and HLA frequencies derived from the HLA phenotypes of more than 12,000 donors recently entered into the US OPTN registry

  34. CPRA Calculator http://optn.transplant.hrsa.gov/ Resources, professional resources, choose cPRA calculator from options

  35. Tambur et al. AJT 2009 Correlation between virtual and Flow crossmatch FP 3.1% Non-HLA abs False pos FCXM FN 14% Some allele specific non-DSA Some weak DSA

  36. Tambur et al. AJT 2009 • Virtual crossmatch is a good tool to predict HLA compatibility. • Caveats: • Antibodies against all donor HLA antigens have to be investigated. • Strength of the antibody has to be considered. • Non-HLA antibodies.

  37. A Virtual Crossmatch Protocol Significantly Increases Access of Highly Sensitized Patients to Deceased Donor Kidney Transplantation. Bingaman et al. Transplantation 2008 FP = 3% 12% Cost effective Decreased TAT Increases access to transplantation of highly sensitized patients

  38. Negative virtual crossmatch predicts negative flow crossmatch Crossmatches performed since implementation of flow crossmatch (June 2010 – September 2011). 157 FP rate = 2.5% 4

  39. Virtual CrossmatchHalifax Lab experience

  40. Renal Transplant Patient Workup • HLA typing, SSO. • Sera collected monthly and after sensitizing event. • Antibody identification by Luminex every 3 months. • Unacceptable antigens and HLA typing are entered into MOTP database. • Donor HLA typing performed and entered into MOTP database. • Smartmatch excludes potential recipients with unacceptable mismatches. • Top 5 potential recipients are selected for crossmatch. • Top 2 recipients with negative crossmatch proceed to Tx • Day of transplant serum and sera collected at 3 weeks and 3 months post transplant are also tested.

  41. Virtual Crossmatch HLA typing VXM HLA antibodies identified

  42. Virtual Crossmatch HLA typing VXM HLA antibodies identified

  43. Virtual Crossmatch HLA typing VXM HLA antibodies identified

  44. Virtual Crossmatch HLA typing VXM HLA antibodies identified

  45. Highly Sensitized Patient Case

  46. 8 8 1 8 1 1 8 1 8 1 1 8 8 8 1 8 1 1 8 1 1 8 1 1 8 8 1 8 8 1 1 8 8 1 8 1 1 8 1 1 1 1 1 1 8 8 8 1 8 8 1 1 8 8 8 1 1 1 1 8 8 1 8 8 1 8 1 1 1 8 1 1 1 8 1 8 1 1 8 8 8 8 1 8 8 8 1 1 1 1 1 8 8 1 8 8 1 8 1 1 8 8 1 1 8 8 8 1 1 8 1 1 8 1 1 1 8 1 8 1 PRA = 95% (55/58) Specificity?

  47. Highly sensitized patient, Case 1 Class I specificity A1 A23 A24 A25 A32 B13 B27 B37 B38 B41 B44 B45 B47 B48 B49 B50 B51 B52 B53 B57 B58 B59 B60 B61 B63 B7 B76 B77 B8 B81 B82 cPRA = 96% • Patient typing A*11,33 B*35,35 Cw*04,04 DRB1*04,13 DR52, 53 DQ*03(7),03(8) • Donor typing A*11,03 B*35,62 Cw*04,10 DRB1*04,11 DR52, 53 DQ*03(7),03(8)

  48. Virtual crossmatch in transplantation from live donors

  49. Case 1 Potential recipient Mother Potential donor Son • Recipient HLA typing • A3,3 B7,7 Cw7,7 DR4,15 DQ6,7 • Donor HLA typing • A1,3 B7,8 Cw7,7 DR4,17 DQ2,7

More Related