2-or

2-OR CASE STUDY: WHEN THE VIRTUAL AND ACTUAL CROSSMATCH ARE DISCORDANT. Jessica L. Jackstadt 1 , Donald M. Constantino 1 , Maryanne Mackey 1 , Laurel Gokee 1 , Jennifer Nielsen 1 , Kari Roberts 1 , Robert Jordan 1 , Peter Jindra 1 , David J. Conti 2 , Amy B. Hahn 1 . 1 Transplantation Immunology Laboratory, Albany Medical College, Albany, NY, USA; 2 Section of Transplantation, Division of General Surgery, Dept. of Surgery, Albany Medical College, Albany, NY, USA. Aim: A 58/F/C kidney candidate with negative auto-B XM and CPRA = 35% appeared on a UNOS matchrun. The donor typed as A2,A31;B35;Cw4, lacking the B8, B64, B65, B39, B18,and B78 unacceptable antigens pre- viously identified by both single antigen ELISA and luminex assays. However, the T-AHG CDC XM was positive with 5 months of current sera. The T-3wash and B-37 CDC XMs were negative. Retrospective T and B flow XMs were strong positive. Methods: Frozen T cell panel with AHG showed 26% PRA with no specificity. Cells bearing B8, B64, B65, B39, and/or B18 were mostly negative. By considering ‘‘2’’ rxns, a PRA of 60% with possible weak anti-A3, B7, B58, and A31 was seen. The HLA-A31 was a mismatched donor antigen leading us to conclude that this was a weak alloantibody not detected by solid phase testing. The patient’s spouse typed as HLA- A3,A32;B14,B-,Bw6, accounting for her anti-A3 and B14 alloantibodies. Results: Further examination showed positive XMs with 5 prior deceased donors. Three of these donors had HLA-B7, two of whom also had A3. All of these donors were T XM positive with and without AHG, and one was also B-37 CDC XM positive. The two other donors were positive only by T-AHG and did not have any of the unacceptable antigens seen by solid phase or frozen cell tray. They did both have the B38 antigen, which might be crossreacting with the patient’s anti-B39 antibody. A sixth donor was negative for all CDC crossmatches and had none of the above antigens. Based on Schnaidt et al (Transplantation 2011;92:510) we retested the serum on the luminex single antigen panel following heat inactivation, addition of EDTA, dilu- tion, and removal of IgM to see if a prozone effect or steric blocking was occurring. No additional antibody specificities were uncovered, and none appeared stronger. A C1q assay was negative except for a very weak anti-B44. Conclusions: Thus, using solid phase single-antigen testing for prediction of crossmatch can miss antibod- ies that can cause positive CDC and flow crossmatches. 2 Abstracts / Human Immunology 73 (2012) 1–47

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Aim: A 58/F/C kidney candidate with negative auto-B XM and CPRA = 35% appeared on a UNOS matchrun.The donor typed as A2,A31;B35;Cw4, lacking the B8, B64, B65, B39, B18,and B78 unacceptable antigens pre-viously identified by both single antigen ELISA and luminex assays. However, the T-AHG CDC XM was positivewith 5 months of current sera. The T-3wash and B-37 CDC XMs were negative. Retrospective T and B flow XMswere strong positive.

Methods: Frozen T cell panel with AHG showed 26% PRA with no specificity. Cells bearing B8, B64, B65,B39, and/or B18 were mostly negative. By considering ‘‘2’’ rxns, a PRA of 60% with possible weak anti-A3,B7, B58, and A31 was seen. The HLA-A31 was a mismatched donor antigen leading us to conclude that thiswas a weak alloantibody not detected by solid phase testing. The patient’s spouse typed as HLA-A3,A32;B14,B-,Bw6, accounting for her anti-A3 and B14 alloantibodies.

Results: Further examination showed positive XMs with 5 prior deceased donors. Three of these donorshad HLA-B7, two of whom also had A3. All of these donors were T XM positive with and without AHG, andone was also B-37 CDC XM positive. The two other donors were positive only by T-AHG and did not haveany of the unacceptable antigens seen by solid phase or frozen cell tray. They did both have the B38 antigen,which might be crossreacting with the patient’s anti-B39 antibody. A sixth donor was negative for all CDCcrossmatches and had none of the above antigens. Based on Schnaidt et al (Transplantation 2011;92:510)we retested the serum on the luminex single antigen panel following heat inactivation, addition of EDTA, dilu-tion, and removal of IgM to see if a prozone effect or steric blocking was occurring. No additional antibodyspecificities were uncovered, and none appeared stronger. A C1q assay was negative except for a very weakanti-B44.

Conclusions: Thus, using solid phase single-antigen testing for prediction of crossmatch can miss antibod-ies that can cause positive CDC and flow crossmatches.

2 Abstracts / Human Immunology 73 (2012) 1–47

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