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- Title
Pronase treatment improves flow cytometry crossmatching results.
- Authors
Apithy, M.‐J.; Desoutter, J.; Gicquel, A.; Guiheneuf, E.; Westeel, P.‐F.; Lesage, A.; Piot, V.; Choukroun, G.; Guillaume, N.
- Abstract
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.
- Subjects
FLOW cytometry; TRANSPLANTATION of organs, tissues, etc.; PRONASE; LYMPHOCYTES; IMMUNOGLOBULINS; B cells; MAJOR histocompatibility complex
- Publication
HLA: Immune Response Genetics, 2017, Vol 90, Issue 3, p157
- ISSN
2059-2302
- Publication type
Article
- DOI
10.1111/tan.13073