Pronase treatment facilitates alloantibody flow cytometric and cytotoxic crossmatching in the presence of rituximab

Bearden, Christopher M.; Ágarwal, Avinash Kumar; Book, Benita K.; Sidner, Richard A.; Gebel, Howard M.; Bray, Robert A.; Pescovitz, Mark D.

doi:10.1016/j.humimm.2004.06.001

Cited by 55 publications

(34 citation statements)

References 11 publications

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“…Two techniques have been developed for testing serum containing chimeric anti-CD20 antibody. One involves enzymatic removal of the CD20 molecule [88], and the other involves removing the anti-CD20 antibody with an immunoglobulin specific for the binding region of the antibody [89]. In either case, tests must include a means of determining whether the treatment was effective.…”

Section: Special Circumstancesmentioning

confidence: 99%

Detecting and monitoring human leukocyte antigen–specific antibodies

Leffell

2008

Human Immunology

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Section: Special Circumstancesmentioning

confidence: 99%

Detecting and monitoring human leukocyte antigen–specific antibodies

Leffell

2008

Human Immunology

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“…31 It has also been shown that rituximab interferes with FC-XM, although the false-positive cros smatches disappear after pronase treatment of B cells. 32 Several studies have shown that the sensitivity and specificity of B-cell crossmatches increase when nonspecific IgG binding to lymphocytes is reduced by pronase pretreatment. It has also been shown that rituximab's CD20 target (which is structurally homologous to Fc receptors) is removed by pronase treatment.…”

Section: Yes (T-and B-cells) No Nomentioning

confidence: 99%

Untitled

Desoutter

Apithy²,

Guillaume³

2017

Exp Clin Transplant

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Preformed donor-specific antibodies against human leukocyte antigen can induce antibody-mediated rejection after organ transplant. Hence, future transplant recipients undergo pretransplant screening for preformed antibodies (ie, virtual crossmatch). Subsequently, prospective (analytic) crossmatching is performed using conventional, complementdependent cytotoxicity assays and/or flow cytometrybased methods. The present article reviews factors that must be considered when unexpected, positive, prospective crossmatches are observed. First, the prozone effect caused by the interference of complement or immunoglobulin M must be abrogated by treating the serum with moderate heat, dilution, hypotonic dialysis, EDTA, or dithiothreitol. Second, the physician must check for the presence of potentially interfering autoantibodies (in a context of autoimmune disease or human immunodeficiency virus infection) or therapeutic antibodies (such as rituximab and antithymocyte globulin). In conclusion, knowledge of each assay's technical characteristics will enable the physician to reliably interpret any discrepancies. The reasons for an unexpected, positive, prospective crossmatch must be elucidated before transplant to ensure efficient organ allocation and optimize patient outcomes.

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“…In the presence of Rituxan, B-cell crossmatches will always appear positive, as Rituxan present in recipient serum will bind to donor cells expressing CD20. Recent studies have demonstrated that, rather than removing the antibody, CD20 can be removed from the B-cell surface via proteolytic cleavage with pronase [58]. Thus, under the appropriate conditions (and with the appropriate controls), positive B-cell crossmatches can be interpreted even in the presence of Rituxan.…”

Section: Confounding Issues: Post-transplantation Conundrums and Chalmentioning

confidence: 99%

“…Cells are treated with pronase, a proteolytic enzyme that will digest Fc receptors without affecting the expression of class I or class II antigens [44]. Pronase also digests CD20 (a target of Rituxan) from the surface of B cells [58]. desensitized before undergoing transplantation will not all have good long-term prognoses, with up to 40% of these patients experiencing antibody-mediated rejection, transplant glomerulopathy, and/or graft loss [32][33][34].…”

Section: Introductionmentioning

confidence: 99%