Validity of Flow Cytometry for Cross-Match Evaluation in Clinical Renal Transplantation

Stefoni, Sergio; Nanni-Costa, A.; Buscaroli, Andrea; Borgnino, Luigi Carlo; Iannelli, S.; Raimondi, Concettina; Scolari, Maria; Feliciangeli, G.; Bonomini, V.

doi:10.1159/000186274

Cited by 14 publications

(11 citation statements)

References 9 publications

(11 reference statements)

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“…Until now, this latter category of complement fixing HLA antibodies has gone unrecognized. This may simply be the result of enhanced fluorescence detection by instrumentation rather than by visual inspection similar to that that described by Stefoni et al (18). Alternatively, since all cells were pronase treated we cannot discount the possibility that pronase removed anti-complementary surface proteins such as decay accelerating factor thereby enhancing the cytolytic activity of complement.…”

Section: The Cfcxm Assaymentioning

confidence: 91%

Cytotoxicity and antibody binding by flow cytometry: A single assay to simultaneously assess two parameters

Saw¹,

Bray²,

Gebel³

2008

Cytometry Part B Clinical

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Background: Cell death, as determined by complement-dependent cytotoxicity or antibody binding, as assessed by flow cytometry, are the two main methods histocompatibility laboratories use to identify HLA antibodies. Conventional cytotoxicity assays, even anti-human globulin (AHG) enhanced, are relatively insensitive, subjective and require complement fixation. In contrast, antibody binding assays (e.g.: flow cytometry crossmatch, FCXM) are sensitive, objective, and complement independent. When sera from potential transplant recipients test positive by both methods, most centers would not proceed to transplant unless patients undergo desensitization. However, when the CDC crossmatch is negative but the FCXM positive, how or whether to proceed is controversial. Frequently, both assays are performed.Methods: In this study, we developed a comprehensive procedure that determines cell death, and antibody binding in a single assay: the cytotoxic flow crossmatch (cFCXM). The assay was validated in parallel studies with two other conventional methodologies, namely AHG and FCXM.Results: Using a combination of eight cells and 15 sera, we have determined an optimal method for the simultaneous detection of HLA antibody binding and cytotoxicity using a single platform. Additionally, we have identified three distinct patterns of HLA antibody reactivity: (1) AHG

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Section: The Cfcxm Assaymentioning

confidence: 91%

Cytotoxicity and antibody binding by flow cytometry: A single assay to simultaneously assess two parameters

Saw¹,

Bray²,

Gebel³

2008

Cytometry Part B Clinical

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“…Similarly, for regrafts one would need 357 patients per group to show a 10% survival advantage assuming a current cadaver regraft survival of 264 Sridhar/M unda/Balakrishnan/First Flowcytometric Crossmatching in Renal Allograft Recipients Several small studies have indicated a worse outcome in FCXM-positive patients with regard to the total number of rejection episodes [4,II], overall graft survival [2,5], early rejection episodes [4,11,12] and graft loss [7,11]. The largest group of FCXM-positive and -negative patients has been reported by the UCLA Tissue Typing Laboratory [6,8,15,16]. In their analysis of 1,060 FCXM-negative first cadaver transplants, 202 FCXM-positive first transplants, 134 FCXM-negative second transplants and 59 FCXM-posi tive second transplants, the 3-month, 6-month and 1-year graft survival was significantly lower in the FCXM-positive regrafts (p < 0.01) compared to the other three groups at all time points [15].…”

Section: Discussionmentioning

confidence: 99%

“…Efforts are continuing to identify re cipients at risk for allograft rejection. The sensitive method of flowcytometry has been used to identify patients with antibodies to donor lymphocytes [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16]. Most studies indi cate a poor outcome in regrafts when the recipient is posi tive by flowcytometric crossmatch (FCXM) testing.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Flowcytometric Crossmatching in Renal Allograft Recipients

Sridhar

Munda²,

Balakrishnan³

et al. 1992

Nephron

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Forty-four cadaver renal allograft recipients who had flowcytometric cross-match (FCXM) testing and sequential quadruple immunosuppression were studied with respect to the number of rejection episodes and the functional viability of the graft in the first year after transplantation. Fourteen of these patients had antibodies to donor T cells by FCXM. All were negative by conventional crossmatch. Multiple-regression analysis with HLA mismatches, panel-reactive antibody (PRA) percentage, flowcytometric channel shifts and transplant number as independent variables revealed that transplant number and high PRA ( > 50%) impacted (p < 0.05) on serum creatinine at 1 month and 1 year, and graft survival at 1 year. In first transplants, a positive FCXM had no impact on 1-year graft survival rates; however, in retransplants, a positive FCXM and/or high PRA had a significant negative impact on 1-year graft survival. This study indicates that the FCXM should be utilized for retransplant patients, and in patients with a high PRA, in an attempt to improve graft survival for these high-risk recipients.

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“…In [9] and [13] a new flow cytometry method for cross-match evaluation has been proposed. This method allows a better detection of weak positive reactions than the light microscopy.…”

Section: Future Development Of the Subsystemmentioning

confidence: 99%

A Web-based Laboratory Subsystem for Serum Antibody Specificity Analysis in a Laboratory Information System

Cabukovski¹,

Golubovski²

2016

IJCA

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In this paper a web-based laboratory subsystem for serum antibody specificity analysis is presented. This subsystem is an efficient laboratory tool for detection and analysis of serum antibody specificity. It is shown that the subsystem can provide satisfactory way for the serum analysis. General TermsHLA antigens, Antibody specificity, Serum quality, Webbased computer program, Detection, Terasaki's microplate. KeywordsWeb-based laboratory system; serum antibody specificity analysis.

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Validity of Flow Cytometry for Cross-Match Evaluation in Clinical Renal Transplantation

Cited by 14 publications

References 9 publications

Cytotoxicity and antibody binding by flow cytometry: A single assay to simultaneously assess two parameters

Cytotoxicity and antibody binding by flow cytometry: A single assay to simultaneously assess two parameters

Evaluation of Flowcytometric Crossmatching in Renal Allograft Recipients

A Web-based Laboratory Subsystem for Serum Antibody Specificity Analysis in a Laboratory Information System

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