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pISSN 2950-9114 eISSN 2950-9122
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Original Article

J Lab Med Qual Assur 2016; 38(3): 159-163

Published online September 30, 2016

https://doi.org/10.15263/jlmqa.2016.38.3.159

Copyright © Korean Association of External Quality Assessment Service.

Effects of Pronase Treatment on Flow Cytometric Crossmatching

Nuri Lee, Ji Won In, Eun Youn Roh, Sue Shin, and Eun Young Song

Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea

Correspondence to:Eun Young Song
Department of Laboratory Medicine, Seoul National University Hospital, Seoul National University College of Medicine, 101 Daehak-ro, Jongno-gu, Seoul 03080, Korea
Tel: +82-2-2072-0197 Fax: +82-2-3672-3337 E-mail: eysong1@snu.ac.kr

Received: May 6, 2016; Revised: June 30, 2016; Accepted: July 5, 2016

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background: Flow cytometric crossmatching (FCXM) is widely used in hospitals performing solid organ transplantation. Pronase treatment of lymphocytes can increase the sensitivity and specificity of B-cell FCXM. However, it can also affect human leukocyte antigen (HLA) expression and results of FCXM. We treated lymphocytes with various concentrations of pronase and analysed the effect of the treatment on the FCXM results.
Methods: The peripheral blood mononuclear cells isolated from 10 renal transplant donors were treated with three different concentrations of pronase (0.5, 1.0, and 2.0 mg/mL). The effects of pronase on median fluorescence intensity (MFI) values of AB serum (Fcγ receptor), HLA class I and II, and on the MFI ratio of HLA class I and II were analysed.
Results: In B-cell FCXM, the MFI values of AB serum (Fcγ receptor) and HLA class I were significantly decreased by the pronase treatment. The MFI ratio of HLA class II was significantly increased upon treatment with 0.5, 1.0, and 2.0 mg/mL pronase (P<0.05, P<0.01, and P<0.01, respectively). In T-cell FCXM, the MFI ratio of HLA class I was significantly decreased by the pronase treatment (all P<0.01).
Conclusions: When performing FCXM, it is recommended that B-lymphocytes should be treated with 1.0 or 2.0 mg/mL pronase. In the case of T-lymphocytes, pronase treatment should be adopted with caution.

Keywords: B-lymphocytes, Pronase, Flow cytometry, crossmatching

References

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