Abrogating biologics interference in flow cytometric crossmatching

Hum Immunol. 2021 Aug;82(8):574-580. doi: 10.1016/j.humimm.2021.01.016. Epub 2021 Apr 30.

Abstract

The flow cytometric crossmatch is currently the gold standard for evaluating donor and recipient histocompatibility. The assay however does have limitations and is sensitive to false positive reactions resulting from the presence of non-HLA antibodies or therapy related immune biologics. Such false positive reactions can lead to the inappropriate decline of an acceptable donor organ or unnecessary therapeutic intervention. Here we describe the successful validation of anti-idiotype blocking antibodies in prevention of false positive flow crossmatch results caused by biologic therapy. Blocking antibodies specific for the Fab portion of Rituximab and/or Alemtuzumab were incubated with biologic containing patient serum prior to use in flow cytometric crossmatching. Biologic blocking successfully negated false positive crossmatch results with Rituximab (B cell ave. % change = -97%) or Alemtuzumab (T cell ave. % change = -99%, B cell ave. % change = -95%) infused sera respectively. Simultaneous blocking of these biologics was also successful. A complex case is presented to demonstrate the application of this procedure.

Keywords: Alemtuzumab; Biologic; Flow cytometric crossmatch; Neutralization; Rituximab.

MeSH terms

  • Antibodies, Blocking / blood
  • Antibodies, Blocking / immunology
  • Antibodies, Neutralizing / blood
  • Antibodies, Neutralizing / immunology
  • Biological Products
  • Dose-Response Relationship, Immunologic
  • Flow Cytometry / methods*
  • Flow Cytometry / standards
  • HLA Antigens / genetics*
  • HLA Antigens / immunology*
  • Histocompatibility / immunology*
  • Histocompatibility Testing / methods*
  • Histocompatibility Testing / standards
  • Humans
  • Isoantibodies / blood
  • Isoantibodies / immunology
  • Neutralization Tests
  • Tissue Donors
  • Transplant Recipients

Substances

  • Antibodies, Blocking
  • Antibodies, Neutralizing
  • Biological Products
  • HLA Antigens
  • Isoantibodies