• Shock · Jun 2021

    Toward Monocyte HLA-DR Bedside Monitoring: A Proof of Concept Study.

    • Pénélope Bourgoin, Ramazan Taspinar, Morgane Gossez, Fabienne Venet, Benjamin Delwarde, Thomas Rimmelé, Pierre-Emmanuel Morange, Fabrice Malergue, and Guillaume Monneret.
    • Department of Research and Development, Beckman Coulter-Immunotech, Marseille, France.
    • Shock. 2021 Jun 1; 55 (6): 782-789.

    ObjectivesDecreased expression of human leukocyte antigen-DR on monocytes (mHLA-DR) is recognized as the most appropriate marker for the monitoring of immune alterations in septic patients and critically ill subjects. Its measurement has been established for years by flow cytometry, but remains under-used due to pre-analytical constraints. The objectives of the present work were to develop a rapid and robust one-step protocol.MethodsA novel, simplified protocol has been developed to measure mHLA-DR in whole blood using flow cytometry. It is a one-step procedure that includes red cell lysis, antibodies, and fixative reagents. It has been compared to the standardized routine protocol in two consecutive cohorts of septic shock patients (n = 37). Finally, the protocol was applied to a few subjects in point-of-care settings, by collecting capillary blood from fingerpricks.ResultsStrong correlation was observed between the one-step method and routine protocol in 24 patients. After testing several stabilizing agents, the procedure was further optimized by adding a low-dose formaldehyde to the stain and lyse solution. This improved method was tested in a second cohort of 13 patients, and again strongly correlated to the routine protocol. Finally, the fingerprick and venous puncture samples were shown to provide similar results.ConclusionsThe present work demonstrates the feasibility of a bedside protocol for flow cytometry measurement of mHLA-DR in critically ill subjects. This helps overcome pre-analytical constraints previously identified, which have limited wider use of this biomarker in intensive care units. In addition, preliminary results from fingerprick samples are promising.Copyright © 2020 by the Shock Society.

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