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Comparison of capillary and venous blood in the analysis of concentration and function of leucocyte sub-populations

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Abstract

Purpose

Compare capillary and venous blood in the analysis of concentration and function of leucocyte sub-populations. This study hypothesised that capillary samples may be used in a site-specific manner as an alternative source of blood samples for assays of leucocyte concentration and neutrophilic phagocytic function and reactive oxygen species (ROS) production, allowing acquisition of multiple samples to better monitor transient but significant post-exercise immune modulation.

Methods

Resting blood samples were simultaneously obtained from vein, finger and earlobe of healthy subjects (n = 10, age: 25.1 ± 3.1 years). Leucocyte concentrations were measured using a five-part differential haematological analyser. Leucocyte sub-populations (CD3, CD4, CD8, CD19, CD56, CD14) and granulocytic functional-related (CD11b, CD18, CD16b, CD66b) surface antigen markers, neutrophil phagocytosis (FITC-labelled Escherichia coli) and stimulated ROS production (DHR) were quantified utilizing flow cytometry. A MANOVA (α < 0.05 significance) analysed the effects of the different sampling sites in the concentrations of leucocyte populations, their surface antigen expression and granulocytic functions.

Results

Leucocyte concentration and neutrophilic ROS production yielded non-significant differences between sampling sites. Expression of granulocytic surface antigens was increased in both capillary sites compared to venous site (p = 0.008), particularly for adhesion markers CD11b/CD18. The percentage of neutrophils performing phagocytosis was higher in venous samples compared to finger (p = 0.025). Increased number of E. coli ingested was observed in venous sample compared to finger (p = 0.001) and to earlobe (p = 0.006).

Conclusion

Whilst attention must be paid for varying neutrophilic surface antigen expression and further studies are needed to establish appropriate reference ranges, this study supports the use of capillary blood samples in a site-specific manner to enhance sampling capabilities field-based research.

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Abbreviations

APC:

Allophycocyanin

APC-Cy™7:

APC-cyanine tandem fluorochrome

APC-H7:

APC-cyanine H7 tandem fluorochrome

CGRP:

Calcitonin-gene-related peptide

CD:

Cluster of Differentiation

DHR:

Dihydrorhodamine

E. coli :

Escherichia coli

EDTA:

Ethylenediaminetetraacetic acid

FITC:

Fluorescein isothiocyanate

fMLP:

Formyl-methionyl-leucyl-phenylalanine

ICAM1:

Intercellular adhesion molecule 1

LPS:

Lipopolysaccharide

MCH:

Mean corpuscular haemoglobin

MCHC:

Mean corpuscular haemoglobin concentration

MCV:

Mean corpuscular volume

MANOVA:

Multivariate analysis of variance

NA:

Neutrophil antigen

PE:

R-phycoerythrin

PE-Cy7:

PE-cyanine tandem fluorochrome

PerCP-Cy™5.5:

PerCP-cyanine tandem fluorochrome

PMA:

Phorbol 12-myristate 13-acetate

PBS:

Phosphate-buffered saline

PNC:

Platelet-neutrophil complexes

ROS:

Reactive oxygen species

RDW:

Red cell distribution width

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Acknowledgments

The authors would like to thank Ms. Evelyne Rathbone for the consultation provided towards the statistical analysis of the data.

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Authors and Affiliations

  1. Faculty of Health Sciences and Medicine, Bond University, 14 University Drive, Robina, 4226, QLD, Australia

    Elisa F. D. Canetti, J. Keane, C. P. McLellan & A. B. Gray

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  1. Elisa F. D. Canetti
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  2. J. Keane
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  3. C. P. McLellan
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  4. A. B. Gray
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Correspondence to Elisa F. D. Canetti.

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Conflict of interest

The authors declare no conflicts of interest.

Ethical standards

Any details that could disclose the identity of subjects were omitted. All procedures in this study were previously approved by the Bond University’s Human Research Ethics Committee (RO1721) and have been performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

Informed consent

Written informed consent was obtained from all individual participants included in the study.

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Communicated by Fabio Fischetti.

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Canetti, E.F.D., Keane, J., McLellan, C.P. et al. Comparison of capillary and venous blood in the analysis of concentration and function of leucocyte sub-populations. Eur J Appl Physiol 116, 1583–1593 (2016). https://doi.org/10.1007/s00421-016-3413-z

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  • DOI: https://doi.org/10.1007/s00421-016-3413-z

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