Profiling antibodies to class II HLA in transplant patient sera

McMurtrey, Curtis P.; Lowe, David; Buchli, Rico; Daga, Sunil; Royer, Derek J.; Humphrey, Alisha; Cate, Steven; Osborn, Sean; Mojsilovic, Aleksandar; VanGundy, Rodney S.; Bardet, Wilfried; Duty, Andrew; Mojsilovic, Danijela; Jackson, Kenneth W.; Šťastný, Peter; Briggs, David; Zehnder, Daniel; Higgins, Rob; Hildebrand, William H.

doi:10.1016/j.humimm.2013.11.015

Cited by 11 publications

(10 citation statements)

References 34 publications

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“…Furthermore, the measured MFI represents only the fraction of isotypes that are targeted by the applied detection antibody conjugate. Indeed, coexistence of IgM and IgA DSA has been described . In an own evaluation of 26 renal allograft recipients having only one HLA class I IgG DSA, we found coexisting IgM DSA in 31% and IgA DSA in 33%, respectively (unpublished data).…”

Section: Technical Challenges and Limitationsmentioning

confidence: 51%

“…In an own evaluation of 26 renal allograft recipients having only one HLA class I IgG DSA, we found coexisting IgM DSA in 31% and IgA DSA in 33%, respectively (unpublished data). Independent of their clinical relevance, these antibodies—not monitored by the standard anti‐IgG pan —may have an influence on MFI, as they are competing for binding on SAB and thus may decrease IgG‐derived MFI . Moreover, the final level of MFI is influenced by concentration, affinity, and incubation time of the used detection antibody conjugate .…”

Section: Technical Challenges and Limitationsmentioning

confidence: 99%

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Caveats of HLA antibody detection by solid‐phase assays

2019

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All authors contributed equally to the review SUMMARY Solid-phase assays for human leukocyte antigens (HLA) antibody detection have clearly revolutionized the field of HLA diagnostics and transplantation. The key advantages are a high sensitivity and specificity for detection of HLA antibodies compared with cell-based assays, as well as the potential for standardization. Solid-phase assays enabled the broad introduction of tools such as "virtual crossmatching" and "calculated panel reactive antibodies," which are essential components in many organ allocation systems, kidney-paired donation programs, and center-specific immunological risk stratification procedures. The most advanced solid-phase assays are the socalled single antigen beads (SAB). They are available now for more than 15 years, and the transplant community embraced their significant advantages. However, SAB analysis and interpretation is complex and many pitfalls have to be considered. In this review, we will discuss problems, limitations, and challenges using SAB. Furthermore, we express our wishes for improvements of SAB as well as their future use for immunological assessment and research purposes.

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Section: Technical Challenges and Limitationsmentioning

confidence: 51%

Section: Technical Challenges and Limitationsmentioning

confidence: 99%

Caveats of HLA antibody detection by solid‐phase assays

2019

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“…). The cells were then treated with anti‐HLA‐I or anti‐HLA‐II at concentrations that ranged from 0.1 to 2 μg mL −1 , which are expected physiologic concentrations of circulating DSAs in patients .…”

Section: Resultsmentioning

confidence: 99%

“…1). The cells were then treated with anti-HLA-I or anti-HLA-II at concentrations that ranged from 0.1 to 2 lg mL À1 , which are expected physiologic concentrations of circulating DSAs in patients [24]. Upon stimulation with anti-HLA-I, TBM expression increased in a dose-dependent manner up to 1 lg mL À1 ( Fig.…”

Section: Endothelial Tbm Surface Expression Following Anti-hla Stimulmentioning

confidence: 95%

Effects of alloantibodies to human leukocyte antigen on endothelial expression and serum levels of thrombomodulin

Béland

Vallin

Désy

et al. 2017

Journal of Thrombosis and Haemostasis

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Rationale Thrombomodulin (TBM) is an anticoagulant and anti-inflammatory transmembrane protein expressed on endothelial cells. Donor-specific alloantibodies, particularly those against human leukocyte antigen (HLA) class II, are associated with microvascular endothelial damage in solid allografts. Objective Our aim was to characterize the effects of anti-HLA antibodies on endothelial expression of TBM, and in particular, the differential effects of anti-HLA class I compared with those of anti-HLA class II. Methods We used human glomerular microvascular endothelial cells to examine TBM expression on anti-HLA-treated cells, and we tested sera from transplant recipients for soluble TBM. Results We found that whereas membrane TBM expression increased in a dose-dependent manner in the presence of anti-HLA class I antibodies, treatment with anti-HLA class II led to minimal TBM expression on the endothelial surface but to a cytosolic accumulation. Platelet adhesion studies confirmed the functional impact of anti-HLA class II. Quantitative densitometry of the membrane lysates further suggested that anti-HLA class II impairs TBM glycosylation. Furthermore, we found a significant association between the presence of circulating anti-HLA class II antibodies in transplant recipients and low serum levels of TBM. Conclusion These results indicate that ligation of anti-HLA class I and II antibodies produces different effects on the endothelial expression of TBM and on serum levels of TBM in transplant recipients. Anti-HLA class II antibodies may be associated with a prothrombotic state, which could explain the higher occurrence of microangiopathic lesions in the allograft and the poor outcomes observed in patients with these alloantibodies.

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“…Higher sensitization levels may cause the increase in the length of time spent on the organ waiting list. 22 Therefore, plasma renin activity (PRA) tests determine the organ transplantation probability of the patient whose presensitization level is low and cross-match test is found as negative before transplantation. 23 The methods used for HLA antibody detection has been developed in the last 10 years.…”

Section: Importance Of the Anti-hla Antibodies In Transplantationmentioning

confidence: 99%