Systematic Comparison of Four Cell- and Luminex-Based Methods for Assessment of Complement-Activating HLA Antibodies

Lachmann, Nils; Todorova, Kremena; Schulze, Harald; Schönemann, Constanze

doi:10.1097/tp.0b013e31827b3dc3

Cited by 47 publications

(22 citation statements)

References 24 publications

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“…However, the proportion of C1q+ dnDSAs was consistent with other studies. 15,19 As previously described, 15,17,18,21 we observed a correlation between DSA MFIs and their ability to bind C1q. We defined an MFI threshold that predicted C1q binding with very good sensitivity, specificity, and positive and negative predictive values.…”

Section: Discussionsupporting

confidence: 49%

“…For the Bordeaux cohort, sera containing dnDSAs underwent further testing using a C1q SAFB assay with a protocol slightly modified from Lachmann et al 18 After heat inactivation at 56°C for 30 minutes and centrifugation, serum (18 ml) was spiked with 4 ml of human C1q (4 mg; Quidel, San Diego, CA) and incubated with 2 ml class I or class II SAFB for 20 minutes at room temperature. A biotinylated monoclonal anti-human C1q antibody (clone 3R9/2, Quidel) was added (30 ml of a 1:30 dilution) and incubated for 20 minutes.…”

Section: C1q-binding Assaymentioning

confidence: 99%

“…Several proposed approaches rely on serum dnDSA SAFB mean fluorescence intensity (MFI) strength, 14,15 IgG subclass analysis, 16 or complement-binding ability. 17,18 DSAs are deleterious via different pathways, but complement C4d deposition in peritubular capillaries reflects the central role of complement in AMR endothelial cell aggression. Because C1q is the complement classic pathway's first protagonist, C1q-binding ability could be a hallmark for clinically relevant serum dnDSA.…”

mentioning

confidence: 99%

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Non-Complement–Binding De Novo Donor-Specific Anti-HLA Antibodies and Kidney Allograft Survival

Guidicelli¹,

Guerville²,

Lepreux³

et al. 2016

Journal of the American Society of Nephrology

125

112

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C1q-binding ability may indicate the clinical relevance of de novo donor-specific anti-HLA antibodies (DSA). This study investigated the incidence and risk factors for the appearance of C1q-binding de novo DSA and their long-term impact. Using Luminex Single Antigen Flow Bead assays, 346 pretransplant nonsensitized kidney recipients were screened at 2 and 5 years after transplantation for de novo DSA, which was followed when positive by a C1q Luminex assay. At 2 and 5 years, 12 (3.5%) and eight (2.5%) patients, respectively, had C1q-binding de novo DSA. De novo DSA mean fluorescence intensity .6237 and .10,000 at 2 and 5 years, respectively, predicted C1q binding. HLA mismatches and cyclosporine A were independently associated with increased risk of C1q-binding de novo DSA. When de novo DSA were analyzed at 2 years, the 5-year death-censored graft survival was similar between patients with C1q-nonbinding de novo DSA and those without de novo DSA, but was lower for patients with C1q-binding de novo DSA (P=0.003). When de novo DSA were analyzed at 2 and 5 years, the 10-year death-censored graft survival was lower for patients with C1q-nonbinding de novo DSA detected at both 2 and 5 years (P,0.001) and for patients with C1q-binding de novo DSA (P=0.002) than for patients without de novo DSA. These results were partially confirmed in two validation cohorts. In conclusion, C1q-binding de novo DSA are associated with graft loss occurring quickly after their appearance. However, the long-term persistence of C1q-nonbinding de novo DSA could lead to lower graft survival.

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Section: Discussionsupporting

confidence: 49%

Section: C1q-binding Assaymentioning

confidence: 99%

mentioning

confidence: 99%

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Non-Complement–Binding De Novo Donor-Specific Anti-HLA Antibodies and Kidney Allograft Survival

Guidicelli¹,

Guerville²,

Lepreux³

et al. 2016

Journal of the American Society of Nephrology

125

112

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“…[21][22][23][24] Two tests have been developed to directly assess the capacity of anti-HLA antibodies to bind the complement components that initiate the classic pathway. [25][26][27][28] The detection of C4d-and C1q-binding anti-HLA antibodies in renal graft recipients correlated with shorter graft survival in small cohorts, [29][30][31][32][33] a result just confirmed for C1q-binding assay in a large cohort by Loupy et al 34 While these findings are extremely promising, none of these pioneer studies have evaluated the prognostic value of these tests at the diagnosis of AMR, the actual time when the clinician most needs a risk stratification assay that may guide therapeutic decision making.…”

mentioning

confidence: 99%

Detection of C3d-Binding Donor-Specific Anti-HLA Antibodies at Diagnosis of Humoral Rejection Predicts Renal Graft Loss

Sicard

Ducreux

Rabeyrin

et al. 2015

Journal of the American Society of Nephrology

236

108

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Antibody-mediated rejection (AMR) is a major cause of kidney graft loss, yet assessment of individual risk at diagnosis is impeded by the lack of a reliable prognosis assay. Here, we tested whether the capacity of anti-HLA antibodies to bind complement components allows accurate risk stratification at the time of AMR diagnosis. Among 938 kidney transplant recipients for whom a graft biopsy was performed between 2004 and 2012 at the Lyon University Hospitals, 69 fulfilled the diagnosis criteria for AMR and were enrolled. Sera banked at the time of the biopsy were screened for the presence of donor-specific anti-HLA antibodies (DSAs) and their ability to bind C1q and C3d using flow bead assays. In contrast with C4d graft deposition, the presence of C3d-binding DSA was associated with a higher risk of graft loss (P,0.001). Despite similar trend, the difference did not reach significance with a C1q-binding assay (P=0.06). The prognostic value of a C3d-binding assay was further confirmed in an independent cohort of 39 patients with AMR (P=0.04). Patients with C3d-binding antibodies had worse eGFR and higher DSA mean fluorescence intensity. In a multivariate analysis, only eGFR,30 ml/min per 1.73 m 2 (hazard ratio [HR], 3.56; 95% confidence interval [CI], 1.46 to 8.70; P=0.005) and the presence of circulating C3d-binding DSA (HR, 2.80; 95% CI, 1.12 to 6.95; P=0.03) were independent predictors for allograft loss at AMR diagnosis. We conclude that assessment of the C3d-binding capacity of DSA at the time of AMR diagnosis allows for identification of patients at risk for allograft loss.

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“…A Luminex SAB IgG-teszt során detektált antitesteknek azonban csak egy része képes a komplementkaszkád aktiválására. Az MFI-értékek csak az erősen reaktív antitestek (magas MFI) esetében korrelálnak a komplementkötő kapacitás-sal, és ezért nem lehet megállapítani, hogy a detektált antitestek közül melyek felelősek a graftkárosodásért [41,42] (1. ábra). A teszt során detektált antitestek vitatott klinikai relevanciáját fi gyelembe véve felmerült a szükségessége egy olyan teszt kifejlesztésének, amelynek segítségével, a Luminex-technológia előnyeit kihasználva, detektálhatók a komplementrendszert aktiváló antitestek [43].…”

Section: A Komplementkötő Antitestek Hatása a Grafttúlélésreunclassified

Challenges of the modern antibody diagnostics in kidney transplantation

Wettstein

Szentiványi

2014

Orvosi Hetilap

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A transzplantációs immunológia kiemelt fontosságú területe az antitestmediált rejectio elleni küzdelem, amelynek hátterében a graft által prezentált humán leukocytaantigének ellen képződő antitestek állnak. A szilárd fázisú immunológiai tesztek bevezetésével jelentősen megváltozott az antitestek monitorozásának gyakorlata, aminek jelentősége van a transzplantációt megelőzően és azt követően is. Ennek ellenére hiányosak az ismeretek a teszt során detektált antitestek klinikai relevanciáját tekintve, ezenfelül technikai kihívások és a betegellátás során alkalmazott különböző stratégiák színesítik a képet. A szerzők a transzplantációs antitestmonitorozás laboratóriumi módszereinek egységesí-tésére vonatkozó ajánlásokat és a vesetranszplantált betegek ellátásában javasolt algoritmusokat mutatják be összefog-laló közleményükben. A szilárd fázisú technikák kombinálása a komplementaktiváció kimutatásával ígéretes eszköz-nek mutatkozik. A komplementkötő antitestek kimutatásán alapuló módszerek fontos kérdésekre adhatnak választ a detektált antitestek klinikai relevanciáját illetően. Az új módszerek alkalmazása során felmerülő kérdések megválaszo-lása és az ajánlások ismerete szükséges ahhoz, hogy ezen a dinamikusan fejlődő területen naprakészek maradjunk. Orv. Hetil., 2014, 155(46), 1820-1830. Kulcsszavak: antitestmediált rejectio, HLA-antitestek, vesetranszplantáció Challenges of the modern antibody diagnostics in kidney transplantationOvercoming antibody mediated rejection is of increasing interest in the fi eld of transplantation immunology. The recipient's antibodies against the graft human leukocyte antigens are responsible for antibody mediated graft injury. Introduction of the solid phase immunoassay technology radically changed the monitoring practice of antibodies against human leukocyte antigens, and this has consequences both for pretransplant and posttransplant phases, though our knowledge about the clinical interpretation of the detected antibodies is limited. This integrating review reports recommendations and algorithms regarding the management of kidney transplant patients. The detection of complement activation combined with the solid phase techniques is a promising new approach in antibody testing. The C4d and especially the more sensitive C1q methods have the potential to answer pivotal questions about the clinical relevance of antibodies. Answering the questions that the applied new methods raised and reviewing the recommendations are needed to remain up to date with this dynamically developing fi eld.

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Systematic Comparison of Four Cell- and Luminex-Based Methods for Assessment of Complement-Activating HLA Antibodies

Cited by 47 publications

References 24 publications

Non-Complement–Binding De Novo Donor-Specific Anti-HLA Antibodies and Kidney Allograft Survival

Non-Complement–Binding De Novo Donor-Specific Anti-HLA Antibodies and Kidney Allograft Survival

Detection of C3d-Binding Donor-Specific Anti-HLA Antibodies at Diagnosis of Humoral Rejection Predicts Renal Graft Loss

Challenges of the modern antibody diagnostics in kidney transplantation

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