Improved specificity and sensitivity when using pronase-digested lymphocytes to perform flow-cytometric crossmatch prior to renal transplantation

Peter I. Lobo, Ross B. Isaacs, Clint E. Spencer, Timothy L. Pruett, Hillary A. Sanfey, Robert G. Sawyer, Christopher McCullough

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Several laboratories have resorted to flow-cytometric cross-match (FCXM) in an effort to prevent hyperacute and accelerated renal allograft rejections. The currently employed FCXM has problems with both false-positive and -negative reactions, largely as a result of irrelevant IgG binding to Fc IgG receptors. In 1980, we circumvented this problem by digesting Fc IgG receptors with pronase, and demonstrated that, with immunofluorescence microscopy (IF), detection of IgG anti-HLA antibodies was highly sensitive and specific. In 1995, we introduced the pronase technique to FCXM and showed that this enzyme did not decrease HLA expression. We present herein a prospective study at our institution to determine whether FCXM using pronase-digested (PD) lymphocytes is as sensitive and more specific than FCXM with undigested (UD) lymphocytes when compared with the highly sensitive and specific IF assay. In analyzing the 186 donor-specific prerenal-transplant crossmatches, we found that PD FCXM was as sensitive and specific as IF and was able to detect weak IgG anti-HLA antibodies that bound to B cells. Fourteen of these patients would have been denied transplants il one were to have relied on UD FCXM. The data clearly indicate that PD FCXM can reliably be used to detect weak IgG anti-HLA antibodies before renal transplantation.

Original languageEnglish (US)
Pages (from-to)563-569
Number of pages7
JournalTransplant International
Volume15
Issue number11
DOIs
StatePublished - 2002

Keywords

  • Anti-HLA antibodies
  • Flow-cytometric crossmatch
  • Pronase
  • Renal allografts Introduction
  • Transplant crossmatch

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