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Article Dans Une Revue HLA: Immune Response Genetics Année : 2017

Pronase treatment improves flow cytometry crossmatching results

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Résumé

Flow cytometry crossmatching (FC-XM) is the most sensitive cell-based method for detecting donor-specific antibodies in clinical organ transplantation. Unfortunately, background FC-XM reactivity is elevated in assays with B lymphocytes-partly because of nonspecific immunoglobulin binding by Fc receptors and B-cell surface immunoglobulins. To reduce the background reactivity in a B-cell FC-XM assay, we treated lymphocytes with pronase (1 mg/mL for 30 minutes). This treatment drastically reduced the presence of kappa light chains and Fc receptors (CD32b), while the concomitant decrease in CD19, CD20 and major histocompatibility complex (MHC) I and II expression on B-cells was acceptable. Higher pronase concentrations (>2 mg/mL) started to significantly affect CD19, CD20, MHC-I and -II expression on B-cells. In subsequent prospective experiments (on 42 donor cells tested with 102 sera), we found that pronase treatment was associated with a relative increase of the sensitivity and specificity in our B-cell FC-XM assay.
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Dates et versions

hal-03574251 , version 1 (15-02-2022)

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M. -J. Apithy, J. Desoutter, A. Gicquel, E. Guiheneuf, P. -F. Westeel, et al.. Pronase treatment improves flow cytometry crossmatching results. HLA: Immune Response Genetics, 2017, 90 (3), pp.157-164. ⟨10.1111/tan.13073⟩. ⟨hal-03574251⟩
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