An Easy and Sensitive Method to Profile the Antibody Specificities of HLA–specific Memory B Cells

Karahan, Gonca E.; Krop, Juliette; Wehmeier, Caroline; Vaal, Yvonne J.H. de; Langerak-Langerak, Janneke; Roelen, Dave L.; Lardy, Neubury M.; Bemelman, Fréderike J.; Berge, Ineke J. M. ten; Reinders, Marlies E.J.; Kooten, Cees van; Claas, Frans H.J.; Heidt, Sebastiaan

doi:10.1097/tp.0000000000002516

Cited by 37 publications

(43 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In case of a de novo immune response, the load of mismatched epitopes—and even more their individual immunogenicity—might be the driving force for rejection. Novel “memory” assays and sophisticated software algorithms (i.e., HLA Matchmaker, PIRCHE II) have been developed to address these gaps . All these tools and their further improvement rely heavily on SAB analyses.…”

Section: The Future Of Sabmentioning

confidence: 99%

Caveats of HLA antibody detection by solid‐phase assays

2019

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Section: The Future Of Sabmentioning

confidence: 99%

Caveats of HLA antibody detection by solid‐phase assays

2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, the utility of these assays is limited by the challenge of developing ELISPOT assays for the array of HLA alleles encountered in the clinic, and the duration of in vitro culture. Another group combined the culture of polyclonal B cells with supernatant analyses using single‐antigen bead (SAB) assay, yielding greater sensitivity in anti‐HLA detection . These studies confirm that donor‐reactive B cells can be detected in sensitized patients, although more work is required to more fully assess their practicality and clinical utility.…”

Section: Memory B Cells and Acute Amrmentioning

confidence: 93%

Outstanding questions in transplantation: B cells, alloantibodies, and humoral rejection

Chong

Rothstein

Safa

et al. 2019

American Journal of Transplantation

View full text Add to dashboard Cite

Over the past three decades, improved immunosuppression has significantly reduced T cell–mediated acute rejection rates, but long‐term graft survival rates have seen only marginal improvement. The cause of late graft loss has been under intense investigation, and chronic antibody‐mediated rejection (AMR) has been identified as one of the leading causes, thus providing a strong rationale for basic science investigation into donor‐specific B cells and antibodies in transplantation and ways to mitigate their pathogenicity. In 2018, the American Society of Transplantation launched a community‐wide online discussion of Outstanding Questions in Transplantation, and the topic of B cell biology and donor‐specific antibody prevention emerged as a major area of interest to the community, leading to a highly engaged dialogue, with comments from basic and translational scientists as well as physicians (http://community.myast.org/communities/community-home/digestviewer). We have summarized this discussion from a bedside to bench perspective and have organized this review into outstanding questions within the paradigm that AMR is a leading cause of graft loss in the clinic, and points of view that challenge aspects of this paradigm. We also highlight opportunities for basic and translational scientists to contribute to the resolution of these questions, mapping important future directions for the transplant research field.

show abstract

“…Among the described cocktails, stimulation with Toll‐like receptor 7/8 agonist (Resiquimod‐R848) and interleukin 2 has been shown to preferentially stimulate memory B cells among peripheral blood mononuclear cells without causing isotype switching in the naïve B‐cell population (Karahan et al., 2017; Pinna, Corti, Jarrossay, Sallusto, & Lanzavecchia, 2009) and is, therefore, increasingly applied. The second approach of HLA‐specific memory B cells analysis is based on profiling of memory B‐cell‐derived HLA antibodies in culture supernatant produced upon such polyclonal stimulation by using luminex SAB assays (Bernasconi et al., 2002; Han, Rogers, Lavingia, & Stastny, 2009; Karahan et al., 2019) (Figure 1, middle row).…”

Section: Hla‐specific Memory B‐cell Assays: Which One To Choose?mentioning

confidence: 99%

“…Importantly, since ELISpot assays rely on the capacity of activated B cells to produce antibodies, viability of the stimulated cells at the end of the pre‐culture phase plays a major role for the quality and accuracy of the results. Hence, in contrast to supernatant analysis favouring longer culture periods up to 10–12 days to achieve accumulation of IgG in the culture supernatants (Karahan et al., 2019), cells to be used for ELISpot assays are generally cultured for 6–7 days in order to obtain an optimum of antibody‐producing viable cells (Heidt et al., 2012; Karahan et al., 2014, 2017). Following culturing, in most protocols activated B cells are transferred to anti‐IgG‐coated ELISpot plates and incubated overnight.…”

Section: Hla‐specific Memory B‐cell Assays: Which One To Choose?mentioning

confidence: 99%

“…Importantly, both tetramer staining and ELISpot assays are very labour‐intensive methods yielding unfamiliar read‐outs for the transplantation community, which render them virtually impossible to use when comparison with serum HLA antibody profiles is desired. In this regard, analysis of polyclonally activated B‐cell culture supernatants for the presence of HLA antibodies using luminex SAB assay allows for such a comparison at similar sensitivity to serum analysis, particularly when IgG is not only concentrated but also affinity‐purified from culture supernatants (Karahan et al., 2019). In addition to being a qualitative assay, supernatant analysis by luminex is also restricted by the HLA specificities included in SAB panels.…”

Section: Hla‐specific Memory B‐cell Assays: Which One To Choose?mentioning

confidence: 99%

See 1 more Smart Citation

HLA‐specific memory B‐cell detection in kidney transplantation: Insights and future challenges

Wehmeier

Karahan

Heidt

2020

Int J Immunogenetics

Self Cite

View full text Add to dashboard Cite

Humoral alloimmunity mediated by anti‐human leucocyte antigen (HLA) antibodies is a major challenge in kidney transplantation and impairs the longevity of the transplanted organ. The immunological risk of an individual patient is currently mainly assessed by detection of HLA antibodies in the serum, which are produced by long‐lived bone marrow‐residing plasma cells. However, humoral alloimmunity is complex, and alloreactive memory B cells constitute an additional factor in the interplay of immune cells. These recirculating “silent” cells are responsible for the immunological recall response by differentiating into antibody‐producing cells upon antigen re‐encounter. Historically, due to the lack of appropriate and routinely applicable assays to determine the presence and HLA specificity of alloreactive memory B cells, their contribution to the humoral alloimmune response has clinically often been suspected but could not be determined. In this review, we give an overview of recent advances in techniques to detect alloreactive memory B cells and discuss their strengths and limitations. Furthermore, we summarize experiences with these techniques in alloimmunized individuals and transplant recipients, thereby emphasizing unmet needs to be addressed in future studies.

show abstract

An Easy and Sensitive Method to Profile the Antibody Specificities of HLA–specific Memory B Cells

Abstract: Supplemental Digital Content is available in the text.

Cited by 37 publications

References 21 publications

Caveats of HLA antibody detection by solid‐phase assays

Caveats of HLA antibody detection by solid‐phase assays

Outstanding questions in transplantation: B cells, alloantibodies, and humoral rejection

HLA‐specific memory B‐cell detection in kidney transplantation: Insights and future challenges

Contact Info

Product

Resources

About