4/22/2015. Platelet antibody screen and crossmatch. HLA antibody testing methods. Matched platelet timeline. Lymphocytotoxicity. Luminex technology

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Esmond Richardson
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$Managing platelet refractory patients: case studies Presented by Paul F. Lindholm, M.D.$

1 Managing platelet refractory patients: case studies Presented by Paul F. Lindholm, M.D. Platelet transfusion Thrombocytopenia/transfusion goals Prophylactic >10K/mL to prevent serious bleeding (prophylactic) Therapeutic >40-50K/mL to treat bleeding or before invasive procedure >75-100K/mL for CNS bleeding/procedure 2 Patient with multiple myeloma Received allogeneic stem cell transplant and later developed sepsis and engraftment failure. Platelet transfusion goal: > 20,000/mL for bleeding Corrected count increments were less than 5,000 M 2 /ml following 2 platelet transfusions; 9 poor at hours. Platelet antibody screen positive 10/13 cells Compatible units obtained with testing: 4/6, 8/12, 6/6, 6/20 HLA antibody screen non-reactive HLA and crossmatched platelets provided. 1

Platelet antibody screen and crossmatch Immucor, Inc.

Matched platelet timeline Provide ABO compatible

Perform platelet crossmatch Obtain HLA class I A&B

2 Platelet antibody screen and crossmatch Immucor, Inc. HLA antibody testing methods Lymphocytotoxicity Luminex technology Medscape.com N Hellman,Renal Fellow Network, Matched platelet timeline Provide ABO compatible platelets Obtain HLA & platelet antibody screen Perform platelet crossmatch Obtain HLA class I A&B typing Request HLA matched platelets Provide XM platelet Collect HLA matched product Provide product 2

3 Platelet count (x10 3 /ml) 4/22/2015 SDP CCI = 18,100 CCI = 5,600 CCI = 8,400 Platelet units: XMC: 6/6 4/ Platelet refractoriness Immune refractoriness Majority HLA Class I A and B Platelet-specific antibodies ABO compatibility Patients may have both immune and nonimmune factors Non-immune refractoriness Splenomegaly (sequestration) Sepsis Fever Bleeding DIC Medications Venoocclusive disease Graft-versus-host disease Platelet storage age 8 Factor Clinically important factors affecting transfusion outcomes 1 hour platelet increment/ml 18 to 24 hour platelet increment/ml Days to next transfusion Overall response 24,900 12, Clinically Important change 5,000 2, Improved platelet response Splenectomy + 24, ,400 - ABO compatible +4,600 +6,300 - Decreased platelet response Lymphocytotoxic antibody* -9,300-4, Female 2 pregnancies* or male -8,900-5, Palpable spleen -3,500-4, Heparin* - -3, Bleeding* -1,700-3, Fever* -1,600-2, Amphotericin B -2,700-2, DIC * Factor also affected platelet refractory rate. SS Slichter et al. Blood 105(10): ,

4 Platelet count (x10 3 /ml) Platelet count 4/22/2015 Platelet increment outcomes Non-refractory Non-immune refractory Immune refractory Patient with end-stage liver disease Patient with cirrhosis, splenomegaly, ascites and hydrothorax developed pancytopenia and refractoriness to platelet transfusions Platelet transfusion goal: > 50,000/mL for bleeding and procedures Corrected count increments were less than 5,000 M 2 /ml following 7 consecutive platelet transfusions Platelet antibody screen positive 10/13 cells 1 platelet unit compatible of 6 tested HLA antibody screen 100% PRA HLA type performed and matched platelets provided CCI = 12,000 CCI = 6,200 CCI = 3,500 Platelet units: XMC: /29 6/46 4/28 7/36 4

5 Summary Our first patient with platelet specific antibodies had one excellent response to crossmatched platelets followed by intermittent modest responses with matched platelets. Our second patient had had highly reactive HLA and platelet antibody screens and obtained initial excellent responses that decreased over time, partly compensated by large platelet doses. Northwestern platelet refractoriness study Retrospective review, 2 years (2012, 2014) Platelet refractoriness studies had both anti-hla and platelet antibody screen/crossmatch Review platelet count increments for all XM or HLA-matched platelet transfusions. Prophylactic response, to 10K/mL Therapeutic, to 40K/mL Best transfusion responses determined for XM/HLA in each case 14 Platelet refractoriness test results Platelet antibody screen HLA Antibody Negative Positive Negative 35 (41%) 15 (18%) Positive PRA > 20% 85 samples in 83 patients No special units needed 12 (14%) HLA-matched units Crossmatch-compatible units 23 (27%) or HLAmatched units 15 5

Percent in Category Corrected Count Increment (CCI) Percent in Category 4/22/2015 Best transfusion responses by unit type 45 10 40 35 30 7 7 8 HLA-matched 8 25 20 4 15 10 5 2 1 0 Therapeutic

6 Percent in Category Corrected Count Increment (CCI) Percent in Category 4/22/2015 Best transfusion responses by unit type HLA-matched Therapeutic Prophylactic Not Therapeutic Not Prophylactic ( 40K/ml) ( 10K/ml) Pre >10 < 40K/ml (not 10K/ml) Patients receiving both HLA and cross-matched platelets Transfusion responses: ABO effect 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% ABO compatible ABO incompatible ABO compatible ABO incompatible Non-T/P Prophylactic Therapeutic

7 Meta-analyses of HLA and crossmatched platelet transfusions HLA: 30 studies selected, I randomized control trial HLA-matched platelets led to better 1-hour post-transfusion platelet count increments than crossmatched or unmatched K Pavenski and ICTMG Collaborators. Transfusion 2013 platelets: 20 studies selected Cross-matched platelets improved platelet count increments versus random or pooled platelets RR Vasallo and ICTMG Collaborators. Transfusion 2014 Effect of matched platelets on bleeding and mortality outcomes not available Studies heterogeneous prospective trials needed Conclusions Almost 60 percent (50/85) of platelet-refractory studies were positive by solid-phase or HLA antibody screen. platelets were readily available and useful for patients with platelet antibodies. HLA-matched platelets tended to yield higher platelet increments. Prophylactic or therapeutic responses were obtained with 61% of ABO compatible matched units versus 36% not receiving ABO compatible. 7

Managing platelet refractory patients: case studies. Presented by Paul F. Lindholm, M.D.

$Managing platelet refractory patients: case studies. Presented by Paul F. Lindholm, M.D.$ Managing platelet refractory patients: case studies Presented by Paul F. Lindholm, M.D. Platelet transfusion Thrombocytopenia/transfusion goals Prophylactic >10K/mL to prevent serious bleeding (prophylactic)