Many genetically defined ABO subgroups exhibit characteristic flow cytometric patterns

Hult, Annika K.; Olsson, Martin L.

doi:10.1111/j.1537-2995.2009.02398.x

Cited by 45 publications

(82 citation statements)

References 61 publications

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“…These results of labeling and profile obtained for A weak and A 2 agree with those presented by Hult and Olsson 16 and with the hemagglutination patterns of serological tests. 5 Our results for A 3 are also consistent with the mixed-field …”

supporting

confidence: 91%

“…16 The percentage of labeled RBCs and the labeling profile were similar to ours in both studies, where the authors analyzed ABO antigens stained by a secondary antibody conjugated to organic dyes using an indirect immunoassay. The overall results demonstrated that a direct immunofluorescence assay using our nanotechnology methodology is capable of differentiating ABO RBC groups.…”

supporting

confidence: 73%

“…15,16 However, these studies were usually performed in fixed cells and by indirect fluoroimmunoassays, using secondary antibodies, which can make the experiments more laborious and less specific. Our methodology with CdTe QDs enabled us to quantitatively evaluate antigens on RBC membranes in A 1 , A 2 , B, A 1 B, O, and in some A weak groups both by profile and by labeling efficiency.…”

Section: Cabral Filho Et Almentioning

confidence: 99%

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Blood group antigen studies using CdTe quantum dots and flow cytometry

Filho¹,

Pereira²,

Fernandes³

et al. 2015

IJN

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New methods of analysis involving semiconductor nanocrystals (quantum dots [QDs]) as fluorescent probes have been highlighted in life science. QDs present some advantages when compared to organic dyes, such as size-tunable emission spectra, broad absorption bands, and principally exceptional resistance to photobleaching. Methods applying QDs can be simple, not laborious, and can present high sensibility, allowing biomolecule identification and quantification with high specificity. In this context, the aim of this work was to apply dual-color CdTe QDs to quantify red blood cell (RBC) antigen expression on cell surface by flow cytometric analysis. QDs were conjugated to anti-A or anti-B monoclonal antibodies, as well as to the anti-H ( Ulex europaeus I) lectin, to investigate RBCs of A 1 , B, A 1 B, O, A 2 , and A weak donors. Bioconjugates were capable of distinguishing the different expressions of RBC antigens, both by labeling efficiency and by flow cytometry histogram profile. Furthermore, results showed that RBCs from A weak donors present fewer amounts of A antigens and higher amounts of H, when compared to A 1 RBCs. In the A group, the amount of A antigens decreased as A 1 > A 3 > A X = A el , while H antigens were A X = A el > A 1 . Bioconjugates presented stability and remained active for at least 6 months. In conclusion, this methodology with high sensibility and specificity can be applied to study a variety of RBC antigens, and, as a quantitative tool, can help in achieving a better comprehension of the antigen expression patterns on RBC membranes.

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supporting

confidence: 91%

supporting

confidence: 73%

Section: Cabral Filho Et Almentioning

confidence: 99%

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Blood group antigen studies using CdTe quantum dots and flow cytometry

Filho¹,

Pereira²,

Fernandes³

et al. 2015

IJN

View full text Add to dashboard Cite

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“…The quantification of CR1 copy number was assessed using biotinylated anti-CR1/CD35 mAb J3D3, followed by a sequential labelling with streptavidin-phycoerythrin (PE), biotinylated anti-streptavidin and streptavidin-PEantibodies as described [61], [62].The amount of A and H antigens displayed on RBC surface was determined by flow cytometry using the following monoclonal antibodies: mAb BRIC-145/9W2 (mouse anti-A antigen, IgG1) and mAb MR3-517 (mouse anti-H antigen, IgM), respectively, and confirmed using TransClone Anti-ABO1 (IgM) and TransClone anti-H1 (IgM) murine mAbs (Bio-Rad Laboratories), respectively. Secondary goat anti-mouse IgG or IgM Alexa fluor 488-conjugated (Molecular Probes, Invitrogen) antibodies were subsequently used.…”

Section: Methodsmentioning

confidence: 99%

Structural Basis for the ABO Blood-Group Dependence of Plasmodium falciparum Rosetting

et al. 2012

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The ABO blood group influences susceptibility to severe Plasmodium falciparum malaria. Recent evidence indicates that the protective effect of group O operates by virtue of reduced rosetting of infected red blood cells (iRBCs) with uninfected RBCs. Rosetting is mediated by a subgroup of PfEMP1 adhesins, with RBC binding being assigned to the N-terminal DBL1α 1 domain. Here, we identify the ABO blood group as the main receptor for VarO rosetting, with a marked preference for group A over group B, which in turn is preferred to group O RBCs. We show that recombinant NTS-DBL1α 1 and NTS-DBL1α 1 -CIDR1γ reproduce the VarO-iRBC blood group preference and document direct binding to blood group trisaccharides by surface plasmon resonance. More detailed RBC subgroup analysis showed preferred binding to group A 1 , weaker binding to groups A 2 and B, and least binding to groups A x and O. The 2.8 Å resolution crystal structure of the PfEMP1-VarO Head region, NTS-DBL1α 1 -CIDR1γ, reveals extensive contacts between the DBL1α 1 and CIDR1γ and shows that the NTS-DBL1α 1 hinge region is essential for RBC binding. Computer docking of the blood group trisaccharides and subsequent site-directed mutagenesis localized the RBC-binding site to the face opposite to the heparin-binding site of NTS-DBLα 1 . RBC binding involves residues that are conserved between rosette-forming PfEMP1 adhesins, opening novel opportunities for intervention against severe malaria. By deciphering the structural basis of blood group preferences in rosetting, we provide a link between ABO blood grouppolymorphisms and rosette-forming adhesins, consistent with the selective role of falciparum malaria on human genetic makeup.

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“…Thus, the propositus was suspected to exhibit blood group chimerism rather than to be of subgroup B3. The flow cytometry for the detection of A/B antigens of red blood cells might also be useful in discriminating between chimerism and ABO subgroups [8]. …”

mentioning

confidence: 99%