The flow cytometric crossmatch in perspective

Talbot, David

doi:10.1016/0966-3274(93)90042-7

Cited by 17 publications

(9 citation statements)

References 112 publications

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“…However, in our hands, an increase of ten channels for non-T cells was felt to be significant in detection of IgG anti-HLA antibodies. Such a criterion maintained sensitivity of UD FCXM and paradoxically decreased the false-positive rate (see Table 1) when compared with other studies that have amplified several-fold the MC value for the negative control AB sera [7,8,9,14,16,17,181. Based on data in Fig.…”

Section: Discussionmentioning

confidence: 65%

Improved specificity and sensitivity when using pronase-digested lymphocytes to perform flow-cytometric crossmatch prior to renal transplantation

Lobo¹,

Isaacs²,

Spencer³

et al. 2002

Transplant International

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Several laboratories have resorted to flow-cytometric crossmatch (FCXM) in an effort to prevent hyperacute and accelerated renal allograft rejections. The currently employed FCXM has problems with both false-positive and -negative reactions, largely as a result of irrelevant IgG binding to Fc IgG receptors. In 1980, we circumvented this problem by digesting Fc IgG receptors with pronase, and demonstrated that, with immunofluorescence microscopy (IF), detection of IgG anti-HLA antibodies was highly sensitive and specific. In 1995, we introduced the pronase technique to FCXM and showed that this enzyme did not decrease HLA expression. We present herein a prospective study at our institution to determine whether FCXM using pronase-digested (PD) lymphocytes is as sensitive and more specific than FCXM with undigested (UD) lymphocytes when compared with the highly sensitive and specific I F assay. In analyzing the 186 donor-specific prerenal-transplant crossmatches, we found that PD FCXM was as sensitive and specific as I F and was able to detect weak IgG anti-HLA antibodies that bound to B cells. Fourteen of these patients would have been denied transplants if one were to have relied on UD FCXM. The data clearly indicate that PD FCXM can reliably be used to detect weak IgG anti-HLA antibodies before renal transplantation.

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Section: Discussionmentioning

confidence: 65%

Improved specificity and sensitivity when using pronase-digested lymphocytes to perform flow-cytometric crossmatch prior to renal transplantation

Lobo¹,

Isaacs²,

Spencer³

et al. 2002

Transplant International

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“…Several technical and conceptual concerns have limited the general clinical utility of B cell FCXM. Defining a positive B cell crossmatch has not been standardized because of various factors, including the lack of uniform correlation with graft survival in most B cell positive patients (reviewed in [8]), the potential for the presence of autoreactive antibodies, and the lack of exact definition, in most studies, of the nature of antibodies binding to B cell, and also the lack of extensive correlation between FCXM results and cytotoxicity assays. Several modifications in technique including the use of a highly specific monoclonal anti‐human IgG antibody markedly reduced background problems, thereby allowing reproducible analysis of B cell FCXM.…”

Section: Discussionmentioning

confidence: 99%

The use of positive B cell flow cytometry crossmatch in predicting rejection among renal transplant recipients

Kotb

Russell

Hathaway

et al. 1999

Clinical Transplantation

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We performed retrospective flow cytometry crossmatch (FCXM) on 106 renal graft recipients who were transplanted based on current T cell negative serologic crossmatch. T and B cell FCXMs were performed on current and historical peak reactive post-transplant sera using 1024-channel flow cytometer and the shift in median channel fluorescence (SMCF) over the negative control was calculated. Cut-off values for a positive T and B crossmatch, > 40 and > 80 SMCF, respectively, were determined based on previous retrospective analysis of the data in the context of clinical outcome in our center, and were 1.5 times the standard deviation (SD) above the mean median channel fluorescence (MCF) of normal sera controls. The 1-yr graft survival was 95% for the total group of patients studied, and 87% for the recipients who had a positive T cell FCXM. To focus on the influence of a positive B cell FCXM on the incidence of rejection, primary transplant recipients who had a negative T cell FCXM (n = 81) were studied. Fifteen of 30 (50%) recipients with a positive B cell FCXM experienced at least one rejection episode within the first year. By contrast, only 15 of 51 (29.4%) of patients with a negative B cell FCXM experienced rejection (p = 0.05). The mean B cell SMCF in the group of patients who had no rejections was 45 +/- 59, while that of the group of patients who experienced at least one rejection was 97 +/- 97 (p = 0.012). By comparison, the rejection rate among the retransplant patients was 44.4%, and the mean B cell SMCF in the group with rejection was 94 +/- 75 while it was 5 +/- 7 among retransplant patients who did not have rejection (p = 0.031). Eighty-six percent of sensitized (panel reactivity antibodies (PRA) > 10%) patients who had a B positive/T negative FCXM experienced rejection, compared to 33% (n = 6 out of 16) of the B negative/T negative sensitized patients (p = 0.03). Furthermore, 62% (n = 13 out of 21) of donor-recipient mismatched patients with a B positive/T negative FCXM experienced rejection, compared to 38% (n = 13 out of 35) of patients with T negative/B negative FCXM who were similarly mismatched (p = 0.064). These data demonstrate the value of a positive B cell FCXM for predicting post-transplant rejections particularly when evaluated in the context of prior sensitization and/or DR mismatching. Our results suggest that B cell FCXM may have significant clinical implications, justifying its use in post-transplant management of recipients who have other risk factors of rejection.

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“…These were detected when donor cells were incubated with serum from the recipient. From this discovery Paul Terasaki developed the lymphocytoxic cross-match using small volumes of serum and a low number of cells (Talbot 1993). This assay identified both IgG and IgM antibodies directed against recipient cells.…”

Section: The Introduction Of Crossmatchingmentioning

confidence: 99%

Immunomonitoring of renal transplant recipients in the early posttransplant period by analysis of cytokine gene expression in peripheral blood mononuclear cells

Gibbs¹,

Sadek²,

Cameron³

et al. 2001

Transplantation Proceedings

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The flow cytometric crossmatch in perspective

Cited by 17 publications

References 112 publications

Improved specificity and sensitivity when using pronase-digested lymphocytes to perform flow-cytometric crossmatch prior to renal transplantation

Improved specificity and sensitivity when using pronase-digested lymphocytes to perform flow-cytometric crossmatch prior to renal transplantation

The use of positive B cell flow cytometry crossmatch in predicting rejection among renal transplant recipients

Immunomonitoring of renal transplant recipients in the early posttransplant period by analysis of cytokine gene expression in peripheral blood mononuclear cells

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