title: Virtual crossmatch in kidney transplantation.
abstract: BACKGROUND
  The Luminex Single-Antigen Beads (LSA) assay allows an accurate detection and characterization of preexisting donor-specific antibodies (DSA) in kidney transplant candidates. But the ability of LSA to detect quite low levels of antibodies makes it hard to correctly predict crossmatch results in donor selection. In this study we retrospectively analyzed the accuracy of our virtual crossmatch (v-XM) protocol, which was used for selection of potential kidney transplant recipients, in predicting the results of actual crossmatch (a-XM) in cadaver-donor renal transplantation. We also investigated correlation between negative a-XM results and strength/specificity of preformed DSA.
  
  
  METHODS
  The correlation between negative v-XMs and a-XMs performed in 2007-2012 at the Regional Transplant Center of the Lazio Region, Italy, was analyzed. In carrying out v-XM, the donor HLA molecules against which patients showed LSA-detected DSA with normalized mean fluorescence intensity (MFI)≥5,000 were considered to be "unacceptable DSA," and LSA-DSA showing MFI<5,000 were defined as "acceptable DSA." All cadaver donors had been typed for HLA-A, -B, -DR, and -DQB molecules by sequence-specific primer methods. On the basis of a negative v-XM, we performed 507 a-XMs between serum samples from 256 renal transplant candidates and T/B lymphocytes from 302 cadaver donors with the use of both complement-dependent cytotoxicity (CDC) and flow cytometry (FC) methods.
  
  
  RESULTS
  The v-XM negative results showed good correlation with both CDC and FC a-XMs (97% and 90%, respectively). The sensitivity of v-XM was 100%; this high value was related to the lack of false-negative DSA results. The limited specificity with both techniques (CDC-XM, 74%; FC-XM, 79%) was due to the presence of "acceptable" and/or anti-DQA/DPB DSA in some patient sera used to perform the a-XMs. During the study period, 171 (67%) of the 256 sensitized patients received a kidney transplant: 30% of these had "acceptable DSA" and/or anti-DQA/DPB DSA. No antibody-mediated rejection due to preformed HLA-DSA was observed.
  
  
  CONCLUSIONS
  Our v-XM protocol showed high sensitivity in predicting donor-recipient immunologic compatibility. The results of this study also demonstrated the importance of evaluating DSA strength for implementing v-XM results in the selection of kidney transplant recipients. Moreover, the finding of anti-DQA/DPB DSA, especially in serum samples that gave positive results with the use of both CDC and FC a-XMs, highlights the importance of defining all of the donor HLA molecules to perform an accurate v-XM.
authors.affiliation: Regional Transplant Center, Lazio Region-National Council of Research, IFT UOS di Roma S Camillo, Rome, Italy. Electronic address: antonina.piazza@cnr.it.
authors.forename: A
authors.initials: A
authors.lastname: Piazza
authors.affiliation: Regional Transplant Center, Lazio Region-National Council of Research, IFT UOS di Roma S Camillo, Rome, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Ozzella
authors.affiliation: Regional Transplant Center, Lazio Region-National Council of Research, IFT UOS di Roma S Camillo, Rome, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Poggi
authors.affiliation: Regional Transplant Center, Lazio Region-National Council of Research, IFT UOS di Roma S Camillo, Rome, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Caputo
authors.affiliation: Regional Transplant Center, Lazio Region-National Council of Research, IFT UOS di Roma S Camillo, Rome, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Manfreda
authors.affiliation: Regional Transplant Center, Lazio Region-National Council of Research, IFT UOS di Roma S Camillo, Rome, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Adorno
mesh_terms: D000906:Antibodies / Q000032:analysis; D046148:Donor Selection; D005260:Female; D005434:Flow Cytometry; D006680:HLA Antigens / Q000276:immunology; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D007558:Italy; D016030:Kidney Transplantation; D008297:Male; D008875:Middle Aged; D012189:Retrospective Studies
chemical_list: D000906:Antibodies; D006680:HLA Antigens