1. EFI Meeting Ulm 2009 Teaching session: Blood Sweat and Tears Derek O’Neill and Ilias Doxiadis

2. Program BST Crossmatching and Screening M. Howell / D. O’Neill: Examples from the thoracic transplantation I. Doxiadis: Stem cell transplantation: to do or not? C. Navarrete: Transfusion related graft injury (TRALI) NN: Other than HLA targets for antibodies in transplantation I. Doxiadis: The conundrum of single antigen bead testing M. Arnold: IgG1 and complement activation CDC is an old dated method: let’s stop it –Pro: Derek O’Neill –Con: Ilias Doxiadis

3. Stem cell transplantation some publications Mickelson EM: HLA matching in hematopoietic cell transplantation. Hum Immunol. 2000 61:92-100 Ottinger et al: Positive serum crossmatch as predictor for graft failure in HLA-mismatched allogeneic blood stem cell transplantation. Transplantation. 2002 73(8):1280-5 Narimatsu H: Successful engraftment in crossmatch- positive HLA-mismatched peripheral blood stem cell transplantation after depletion of antidonor cytotoxic HLA antibodies with rituximab and donor platelet infusion. Bone Marrow Transplant. 2005 36:555-6. Conclusion:Higher incidence of graft failure, but not a contraindication

4. Evolution of HLA antibody screening commercialhome made SPA = solid phase assay Evolution of the crossmatch CDC FCM SPA gen SPA sin CDC  FCM  SPA gen (  ) SPA sin

6. Immunoglobulin isotype switch IgM IgG 3 IgA 2 IgG 1 IgE IgD IgA 1 IgG 2 IgG 4 One way street sIgM+IgD+ Spriewald, Arnold C’activation

7. CDC Complement Dependent Cytotoxicity Terasaki PI and McClealand JD: Microdroplet assay of human serum cytotoxins. Nature, 1964: 204:998-1000. NEGPOS

8. Conundrum? co ⋅ nun ⋅ drum Spelled Pronunciation [kuh-nuhn-druhm] 1. a riddle, the answer to which involves a pun or play on words, as What is black and white and read all over? A newspaper. 2. anything that puzzles. Origin: 1590–1600; pseudo-L word of obscure orig.

9. CDC: complement dependent cytotoxicity; ELI: ELISA; LUM: Luminex 75% consensus rule: If 75% of the participants report a specificity than this is regarded as correct; 95% consensus rule: if ≥5% of the participants report a specificity than this is regarded as incorrect Conclusion: the use of two methods lead to better results Example of an External Proficiency Testing exercise

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11. Cardiothoracic Organs Antibody screening & Crossmatching Questions –To crossmatch or not ? CDC/Flow crossmatch ? – timelines involved and organisation Virtual crossmatch? –Antibody screening to support these What methods – what sensitivity? Some examples from two centres

12. Renal/Pancreatic Antibody screening & Crossmatching EFI - requirements Methods – practicalities and cost Minimum or maximum approach? How sensitive is too sensitive ? Crossmatching - cdc or flow or cdc/flow?