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Growth Requirements, Binding and Migration of Human Natural Killer Cells
Published in Ronald H. Goldfarb, Theresa L. Whiteside, Tumor Immunology and Cancer Therapy, 2020
Tuomo Timonen, Juha Jääskeläinen, Anna Mäenpää, Tuula Helander, Anatoly Malygin, Panu Kovanen
NK cells express many adhesion molecules (9) which makes them the most adhesive lymphocyte subpopulation in peripheral blood. The panel of the mAb we used in the present study covers most of the known adhesion receptors. Our results show that the already abundant expression of adhesion structures is on the average upregulated by IL-2. An interesting novel finding is that CD61, the β3 chain of integrins, is strongly upregulated by IL-2. We have not yet been able to identify the alpha chain associated to β3, although immunoprecipitations with anti-β3 antibodies clearly reveal a band that fits to the molecular weight range of the integrin alpha chains. We are currently using alpha chain oligonucleotide probes in order to identify the molecule by Northern blotting.
Immunopathology
Published in Constantin A. Bona, Francisco A. Bonilla, Textbook of Immunology, 2019
Constantin A. Bona, Francisco A. Bonilla
As mentioned above, other cytopenias may also be the result of autoantibody production. Idiopathic thrombocytopenic purpura (ITP) is caused by anti-platelet autoantibodies. These are directed mainly against the surface glycoprotein heterodimer gpIIb/IIIa (CD41/CD61 β3 integrin, see Chapter 10). Autoimmune neutropenia is caused by antibodies reactive with neutrophil surface antigens.
Chronic Myeloid Leukemia
Published in Wojciech Gorczyca, Atlas of Differential Diagnosis in Neoplastic Hematopathology, 2014
Blasts are positive for CD34, CD117, CD33, myeloperoxidase (MPO), and HLA-DR. Promyelocytes are positive for CD117, CD33, and MPO; they do not express CD34 and HLA-DR. The staining with MPO, CD71, and glycophorin A (GPHA) shows marked predominance of myeloid cells. Megakaryocytes are positive for CD61. In cases with increased blasts, immunostaining with terminal deoxynucleotide transferase (TdT), CD34, CD117, myeloid markers, and B- and T-cell markers is helpful to establish the lineage of immature cells and their proportion (e.g., to differentiate between myeloid and lymphoid blast crises).
Evaluating stored platelet shape change using imaging flow cytometry
Published in Platelets, 2023
Tahsin Özpolat, Olga Yakovenko, Anastasiia Stratiievska, S. Lawrence Bailey, Jeffrey Miles, Chomkan Usaneerungrueng, Daire Byrne, Xiaoping Wu, Moritz Stolla
Samples were analyzed by using Amnis Imagestream Mk II (Luminex Corp, Austin, TX). Images of cells were then acquired with the Amnis Image Stream with the 60X magnitude objective using the bright field channel, the SSC channel, FITC (channel 2) and APC (Channel 11) by the INSPIRE software for acquisition. Per sample, 20000 events were recorded on the Imaging Flow Cytometry. IDEAS 6.2 software (Luminexcorp, Austin, TX) were used for analysis. First, we identified focused platelets based on bright-field images by the levels higher than 60 on gradient root mean (GRM) square channel 1 (M01) feature to analyze platelet morphology (Figure 2a). We isolated platelets based on CD61 positivity (we gated for CD61 positive cells based on 1e3-2e5 MFI, Figure 2b). In the third step, we defined single platelets by using the Area and the Aspect Ratio features (Figure 2c). Lastly, we used Aspect Ratio (circular events with a transverse axis of >0.8 of the longitudinal axis, and fusiform events with ≤0.8 of the longitudinal axis) to analyze different morphologies of stored platelets (Figure 2d). The boundaries of the gates were defined based on the shape of circular versus fusiform dot plots. This was further refined and confirmed with image controls. The percent of the circular and fusiform events were used for statistical analysis.
Distinct phenotypes of platelet, monocyte, and neutrophil activation occur during the acute and convalescent phase of COVID-19
Published in Platelets, 2021
Yashuan Chao, Johan Rebetz, Anna Bläckberg, Gisela Hovold, Torgny Sunnerhagen, Magnus Rasmussen, John W. Semple, Oonagh Shannon
Following activation, platelets can bind to circulating leukocytes. Platelet-monocyte complex (PMC) or platelet-neutrophil complex (PNC) formation was determined using flow cytometry of whole blood in the presence or absence of ex vivo stimulation with the platelet agonist thrombin. Platelet-positive events were determined by staining with CD61. There was a significant increase in PMC formation (Figure 4A) and PNC formation (Figure 4C) in response to thrombin stimulation in patients on day 0 (P < .001, PMCs; P = .0003, PNCs) and day 28 (P = .002, PMCs; P = .0017, PNCs) and in healthy controls (P < .001, PMCs; P = .0001, PNCs). As was the case for platelet activation in PRP, Patient 4 failed to generate PMCs or PNCs in response to thrombin stimulation on day 28, and this is likely explained by the observation that this patient had highly elevated PMC and PNC formation already at baseline (without stimulation) on day 28.
Regulation of differentiation of MEG01 to megakaryocytes and platelet-like particles by Valproic acid through Notch3 mediated actin polymerization
Published in Platelets, 2019
Ankita Dhenge, Rutuja Kuhikar, Vaijayanti Kale, Lalita Limaye
Under in vivo conditions, platelets are activated and recruited to the site of blood vessel injury to form aggregates. To check whether culture derived platelets are true platelets, we tested their activation profile after treatment with type I collagen as agonist. This was done by gating CD61 positive viable cells and then analyzing the surface expression of membrane bound Pselectin-CD62P, on the gated cells. As expected, the activation was higher in VPA set on the 21st day (Figure 3c). Fig. S2d shows data from three experiments, and Fig. S2e shows activation of blood platelets that served as a positive control. To summarize, the mature megakaryocytes show platelet shedding from day 14 up to day 21, and this phenomenon was more pronounced in the VPA set. The platelet-like particles were functionally active.