A more efficient preparation system for HLA-eliminated platelets

Vox Sang. 2020 Feb;115(2):159-166. doi: 10.1111/vox.12859. Epub 2019 Nov 25.

Abstract

Background and objectives: Although HLA-eliminated platelets can facilitate transfusions to patients possessing HLA antibodies, no such products are currently available commercially perhaps because the platelet collection rate is not yet economically viable. We have improved this process' efficiency by employing a hollow-fibre system at the last step of the production process after an acid and a reaction buffer have been washed out conventionally by centrifugation.

Materials and methods: HLA-eliminated platelets were prepared via four distinct steps: chilled on ice, treated with an acid solution, diluted and finally washed using the hollow-fibre system. The efficiency of this platelet recovery process was determined. The resulting products' platelet characteristics, including a capacity for HLA expression, were evaluated in vitro and compared in detail to their corresponding originals.

Results: The average efficiency of platelet recovery was 91%. Although the expression levels of CD62P, a molecular marker for platelet activation, were approximately threefold higher on new platelets than on the original platelets, their HLA expression levels were lower. The phagocytosis assay, with monoclonal antibodies and cognate HLA antibody-containing sera, suggested that HLA-ABC molecules on the cell surface were sufficiently removed. The platelet functions, including the agonist-induced aggregability and adherence/aggregability of the collagen-coated plates under certain conditions, were conserved and not significantly different from the original ones.

Conclusion: We propose a novel preparation system for producing HLA-eliminated platelets without centrifugation, which ensures a highly efficient, and therefore, much more economical method of platelet recovery that also retains their key functionality.

Keywords: HLA-eliminated platelets; hollow fibre; platelet.

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Blood Platelets / cytology*
  • Blood Platelets / immunology
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Cell Separation / standards
  • Centrifugation / adverse effects
  • HLA Antigens / immunology
  • Humans
  • P-Selectin / genetics
  • P-Selectin / metabolism
  • Platelet Activation

Substances

  • Antibodies, Monoclonal
  • HLA Antigens
  • P-Selectin
  • SELP protein, human