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. 2014 Nov;99(11):e214-7.
doi: 10.3324/haematol.2014.108555.

Prominent role of platelets in the formation of circulating neutrophil-red cell heterocellular aggregates in sickle cell anemia

Venina M Dominical  1 , Leigh Samsel  2 , James S Nichols  3 , Fernando F Costa  1 , J Phillip McCoy Jr  2 , Nicola Conran  4 , Gregory J Kato  3
Affiliations

Prominent role of platelets in the formation of circulating neutrophil-red cell heterocellular aggregates in sickle cell anemia

Venina M Dominical et al. Haematologica. 2014 Nov.
No abstract available

Keywords: circulating neutrophil-red cell heterocellular aggregates; platelets; sickle cell anemia.

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Figures

Figure 1.
Figure 1.
The formation of circulating neutrophil-RBC heterocellular aggregates is augmented in SCA patients. (A) Percentage of neutrophils (CD66b+) aggregated to red blood cells (RBCs) in peripheral blood from healthy individuals (Control, n=11) and SCA patients on/off HU (SCA, n=23), as detected by imaging flow cytometry. **P<0.01, compared to control group (Mann-Whitney test). (B) Percentage of neutrophils (CD66b+) aggregated to immature (CD235a+/CD71+) and mature (CD235a+/CD71−) RBCs in the peripheral blood of healthy individuals (Control, n=11) and SCA patients on/off HU (SCA, n=23), as detected by imaging flow cytometry; ***P<0.001, compared to respective control group (Mann-Whitney test). (C) Representative brightfield (BF) and fluorescent images, acquired by imaging flow cytometry; CD66b+ neutrophils (purple) aggregated to immature RBCs (CD235a+ - yellow; CD71+ - green) or mature RBCs (CD235a+ - yellow; CD71−); Final image: merged CD66b+/CD235a+/CD71+. Cells are from representative samples of peripheral blood from control, SCA and SCAHU patients. (D) Correlation of reticulocyte counts, (E) Fetal hemoglobin (HbF) levels and (F) platelet counts with percentages of neutrophil-reticulocyte aggregates (CD235a+/CD71+) in SCA (on/off HU) patients (n=17); Spearman’s non-parametric correlation test.
Figure 2.
Figure 2.
Inhibition of Mac-1, VLA-4 and ICAM-4 function reverses the formation of neutrophil-RBC heterocellular aggregates. (A) Representative brightfield (BF) and fluorescent images, acquired by imaging flow cytometry, of CD66b+ neutrophils (purple) expressing CD11a (LFA-1 subunit) and CD11b (Mac-1 subunit) and aggregated to immature RBCs (CD235a+ - yellow color; CD71+ - green) or mature RBCs (CD235a+ - yellow; CD71−); Final image: merged CD66b/CD235a/CD71. Aggregates from the peripheral blood of control, SCA and SCAHU patients. Effects of the incubation of granulocyte suspensions (12 min, 37°C) with a non-specific antibody (IgG1), or adhesion-molecule blocking antibodies/peptides against the CD11a (LFA-1), CD11b (Mac-1), CD49d (VLA-4), ICAM-4 and BCAM molecules, on the percentage of neutrophils aggregated to (B) immature RBCs (CD235+CD71+) and (C) mature RBCs (CD235+CD71−) in samples from SCA patients (n=13). Anti-CD11a [clone 38; 10 μg/mL] and anti-CD11b [clone ICRF44; 10 μg/mL] were from AbD Serotec, Raleigh, NC, USA; anti-CD49d [clone 2B4, 10 μg/mL], and anti-BCAM [25 μg/mL] were from R&D Biosystems, Minneapolis, MN, USA; ICAM-4 blocking peptide (SC-27685, 200 μg/mL) was from Santa Cruz Biotech, Santa Cruz, CA, USA. Non-specific IgG1 (10 μg/mL) was from R&D Biosystems. *P<0.05, **P<0.01 (Friedman/Dunn’s), compared to non-specific IgG1 antibody.
Figure 3.
Figure 3.
Involvement of platelets in SCA neutrophil-RBC interactions. Percentages of neutrophils from steady-state SCA (on/off HU) individuals aggregated to (A) RBCs (CD235a+), (B) reticulocytes (CD235a+/CD71+) and (C) mature RBCs (CD235a+/CD71−) under basal conditions and after incubation of granulocyte suspension with a non-specific IgG antibody or P-selectin function-blocking antibody (Anti-CD62P; clone 9E1, 2 μg/mL) (n=7); *P<0.05, **P<0.01, compared to basal (Friedman/Dunn’s). Percentages of neutrophils aggregated to (D) RBCs (CD235a+), (E) reticulocytes (CD235a+/CD71+) and (F) mature RBCs (CD235a+/CD71−), and demonstrating the presence (+) or not (−) of activated (P-selectin-positive) platelets in these aggregates when analyzed by flow cytometry (SCA on/off HU; n=7). (G) Representative images of the involvement of platelets (CD62P+; red) in the aggregates of neutrophils (CD66b+; purple) with RBC (CD235a+; yellow) expressing, or not, the transferrin receptor (CD71+; green). BF: brightfield mode and respective channels of fluorescence.
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References

    1. Hidalgo A, Chang J, Jang JE, Peired AJ, Chiang EY, Frenette PS. Heterotypic interactions enabled by polarized neutrophil microdomains mediate thromboinflammatory injury. Nat Med. 2009;15(4):384–91. - PMC - PubMed
    1. Turhan A, Jenab P, Bruhns P, Ravetch JV, Coller BS, Frenette PS. Intravenous immune globulin prevents venular vaso-occlusion in sickle cell mice by inhibiting leukocyte adhesion and the interactions between sickle erythrocytes and adherent leukocytes. Blood. 2004; 103(6):2397–400. - PubMed
    1. Turhan A, Weiss LA, Mohandas N, Coller BS, Frenette PS. Primary role for adherent leukocytes in sickle cell vascular occlusion: a new paradigm. Proc Natl Acad Sci USA. 2002;99(5):3047–51. - PMC - PubMed
    1. May AE, Langer H, Seizer P, Bigalke B, Lindemann S, Gawaz M. Platelet-leukocyte interactions in inflammation and atherothrombosis. Semin Thromb Hemost. 2007;33(2):123–7. - PubMed
    1. Gawaz M, Fateh-Moghadam S, Pilz G, Gurland HJ, Werdan K. Platelet activation and interaction with leucocytes in patients with sepsis or multiple organ failure. Eur J Clin Invest. 1995;25(11):843–51. - PubMed

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