Flow cytometry antibody screening using pooled red cells

Won, Dong Il; Jung, Ok-Ju; Lee, Yun Soo; Kim, Sang Geol; Suh, Jang Soo

doi:10.1002/cyto.b.20494

Cited by 5 publications

(5 citation statements)

References 22 publications

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“…Furthermore, antibody titers can be quantified by using advanced methodologies, such as flow cytometry or immunoassays, which could have yielded more accurate correlations [32]. However, further studies with larger cohorts are required to elucidate the influence of ABH secretor status on long-term outcomes.…”

Section: Discussionmentioning

confidence: 99%

Effects of Neutralization by Soluble ABH Antigens Produced by Transplanted Kidneys From ABO-Incompatible Secretor Donors

Kim

Hwang

et al. 2017

Ann Lab Med

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BackgroundGrafts survive despite blood group antigens on the transplant being continuously exposed to antibodies in the blood of recipients in ABO-incompatible kidney transplantation (ABOi KT), owing to the mechanism of accommodation. We analyzed the immunodynamics of soluble ABH antigens in allografts from secretor donors and the influence of such immunodynamics on accommodation and subsequent graft survival in ABOi KT.MethodsThe genotype of a known human β-galactoside α-1,2-fucosyltransferase gene (FUT2), which determines soluble ABH antigen secretor status, was established in 32 donors for ABOi KT at the Severance Hospital, from June 2010 to July 2015. Clinical outcomes of recipients, such as anti-A/B antibody titer change, renal function, and graft survival, were evaluated.ResultsTwenty-five donors were secretors (78.1%), and seven were nonsecretors (21.9%). The frequency of anti-A/B IgG or IgM antibody titer elevation or reduction post-transplantation was not significantly related to donor secretor status. However, IgM titer was rapidly reduced in recipients transplanted from nonsecretor donors (P=0.01), which could be explained by the lack of absorption effect of soluble antigens, enhancing the binding of antibodies to antigens in the allografts. Interestingly, soluble ABH antigens did not affect rejection-free graft survival, which may be due to the nature of β-galactoside α-1,2-fucosyltransferase.ConclusionsSoluble ABH antigens produced by transplanted kidneys from secretor donors played a role in inducing accommodation within three months of KT through neutralization; however, major graft outcomes were not affected.

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Section: Discussionmentioning

confidence: 99%

Effects of Neutralization by Soluble ABH Antigens Produced by Transplanted Kidneys From ABO-Incompatible Secretor Donors

Kim

Hwang

et al. 2017

Ann Lab Med

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“…2). In a given pool, alloantibodies can be suspected to exist based on the percentage of the positive peaks of the total gated cells, according to the assumed number of reactive donors among the total number of donors in the pool (15)(16)(17). First, if a single peak arose, the M1 region was set around its major portion.…”

Section: Discussionmentioning

confidence: 99%

“…This is because the WBC suspensions used for the experiment were pooled from five donors. In a given pool, alloantibodies can be suspected to exist based on the percentage of the positive peaks of the total gated cells, according to the assumed number of reactive donors among the total number of donors in the pool (15)(16)(17). Theoretically, if a single type of reactive alloantibody exists, its peak will occupy at least one-fifth of the total gated cells.…”

Section: Discussionmentioning

confidence: 99%

“…2) was created equidistantly around the position of the modal value for the entire gated cells, the upper and lower ends of which were set to maximally cover the symmetric portion of the peak (13,14). When multiple peaks arose, the M1 region was drawn around one or more positive peaks only (15)(16)(17). The individual MFI ratio for each of the three cell types for a patient were then calculated by dividing the average MFI value of the patient's samples in duplicate by the average MFI value of the six negative controls included in the run.…”

Section: Flow Cytometry Data Analysismentioning

confidence: 99%

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A Novel Flow Cytometric Method for the Simultaneous Detection of Antibodies Against Platelet, Lymphocyte, And Neutrophil

Suh

Won

2016

Cytometry Part B Clinical

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“…We evaluated 100,000 events per sample to show the erythrocyte agglutination levels in the tubes. Agglutination levels were calculated with the single-cell analysis and forward-scatter gating strategy [ 15 ].…”

Section: Methodsmentioning

confidence: 99%

Incomplete Antibodies May Reduce ABO Cross-Match Incompatibility: A Pilot Study

Ozen

Yılmaz

Özkan

et al. 2018

Tjh

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Objective:Any erythrocyte transfusion among humans having type A or B blood groups is impossible due to antibodies causing fatal transfusion complications. A cross-match test is performed to prevent immune transfusion complications before transfusion. Our hypothesis is that the fragment antibody (Fab) part of the antibody (incomplete antibody) may be used to prevent an immune stimulus related to the complete antibody. Therefore, we designed a pilot study to evaluate the effectiveness of these incomplete antibodies using cross-match tests.Materials and Methods:Pepsin enzyme and staphylococcal protein A columns were used to cut anti-A and anti-B monoclonal antibodies and purify their Fab (2) fragments, respectively. An Rh-positive erythrocyte suspension with purified anti-A Fab (2) solution and B Rh-positive erythrocyte suspension with purified anti-B Fab (2) solution were combined correspondingly. Cross-match tests were performed by tube and gel centrifugation methods. The agglutination levels due to the anti-A and anti-B Fab (2) antibodies and their effects on the agglutination normally observed with complete antibodies were then measured.Results:No agglutination for the purified incomplete anti-A Fab (2) with A Rh+ erythrocyte and anti-B Fab (2) with B Rh+ erythrocyte combinations was observed in the tube cross-match tests. These agglutination levels were 1+ in two wells in the gel centrifugation cross-match tests. Fab (2)-treated erythrocytes were also resistant to the agglutination that normally occurs with complete antibodies.Conclusion:We determined that the Fab (2) fragments of antibodies may not only be used to obtain a mild or negative reaction when compared to complete antibodies, but they might also be used for decreasing ABO incompatibility. Incomplete antibodies might be a therapeutic option in autoimmune hemolytic anemia and they may also be used in solid organ or hematopoietic stem cell transplantation. Therefore, we have planned an in vivo study to prove these in vitro findings.

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Flow cytometry antibody screening using pooled red cells

Cited by 5 publications

References 22 publications

Effects of Neutralization by Soluble ABH Antigens Produced by Transplanted Kidneys From ABO-Incompatible Secretor Donors

Effects of Neutralization by Soluble ABH Antigens Produced by Transplanted Kidneys From ABO-Incompatible Secretor Donors

A Novel Flow Cytometric Method for the Simultaneous Detection of Antibodies Against Platelet, Lymphocyte, And Neutrophil

Incomplete Antibodies May Reduce ABO Cross-Match Incompatibility: A Pilot Study

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