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. 2019;115(2):149-155.
doi: 10.1159/000494103. Epub 2018 Nov 27.

Platelet-Neutrophil Interactions Are Lower in Cord Blood of Premature Newborns

Ijeoma Esiaba  1 , Danilyn M Angeles  2 , Terry-Ann M Milford  3 , Lorena M Salto  4 , Kimberly J Payne  3 , Melissa Y Kidder  5 , Danilo S Boskovic  6   7
Affiliations

Platelet-Neutrophil Interactions Are Lower in Cord Blood of Premature Newborns

Ijeoma Esiaba et al. Neonatology. 2019.

Abstract

Objective: To quantify platelet-neutrophil interaction by flow cytometry, in newborn cord blood, as a function of gestational age.

Rationale: Little is known about platelet function markers in the newborn, and developmental variations in these markers are not well described.

Methods: Cord blood samples were obtained from 64 newborns between 23 and 40 weeks' gestation. The neonates were grouped into three categories: preterm (< 34 weeks' gestation, n = 21), late preterm (34 to < 37 weeks' gestation, n = 22), and term (≥37 weeks' gestation, n = 21). We monitored the expression of P-selectin and the formation of platelet-neutrophil aggregates (PNAs) by flow cytometry while using adenosine 5'-diphosphate (ADP) or thrombin receptor-activating peptide (TRAP) as agonists.

Results: PNAs were significantly lower in preterm compared to term neonates after TRAP or ADP stimulations (11.5 ± 5.2% vs. 19.9 ± 9.1%, p < 0.001, or 24.0 ± 10.1% vs. 39.1 ± 18.2%, p = 0.008, respectively). The expression of P-selectin also tended to be lower in preterm neonates, with significant positive correlations between P-selectin expression and PNA formation.

Conclusions: The potential formation of PNAs correlates with gestational age. This suggests that the development of functional competencies of platelets and neutrophils continues throughout gestation, progressively enabling interactions between them.

Keywords: Neutrophil; Neutrophil function; Platelet; Platelet activation; Platelet function; Platelet-neutrophil interaction; Prematurity.

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Conflict of interest statement

Conflict of Interest

The authors declare that they have no conflict of interest.

Figures

Figure 1.
Figure 1.
Gating for platelet-neutrophil aggregates. In the scatter plot at left, neutrophils were gated as the CD45LO and CD16+ double positive cell population. Histograms at right show the percent of platelet-neutrophil aggregates positive for CD61 (right panels) without and with stimulation with ADP (right panel) or TRAP (data not shown). The left histogram shows the fluorescence minus one (FMO) control used to set gates for CD61 positivity.
Figure 2.
Figure 2.
Platelet-Neutrophil Aggregates. Percentage CD61 PerCP positive neutrophils in cord blood of preterm (n=21), late preterm (n=22) and term (n=21) newborns. Results are mean ± SD, analysis based on one-way ANOVA test and Bonferroni multiple comparison. Power for ANOVA analysis of PNA after TRAP and ADP stimulations were .90 and .76 respectively.
Figure 3.
Figure 3.
Pearson correlation of the log % P-Selectin positive platelets with platelet-neutrophil aggregates in the cord blood samples of 64 neonates during (A) TRAP (power=.98) and (B) ADP (power=.99) stimulations.
Figure 4.
Figure 4.
The percentage cord blood platelets expressing P-Selectin in preterm (n=21), late preterm (n=22) and term (n=21) newborns. Results are mean ±SD, analysis based on one-way ANOVA after log transformation. Power = .48 for P-Selectin expression after TRAP stimulation.
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