A PCR-based method for quantifying neutrophils in human nasal secretions

Department

Research

Document Type

Article

Publication Title

Journal of Immunological Methods

Abstract

Neutrophil recruitment to the nasopharynx (NP) is a central event in resolution of NP-initiated microbial infections. A vigorous neutrophil response in infected tissues is also associated with the outcome of adverse tissue pathology. Therefore, differences in infection-induced tissue neutrophil numbers may correlate with pathogenesis events. Existing methods of quantifying neutrophils require evaluation of NP samples within hours of procurement as flow cytometry based cell quantification methods require live neutrophil cells. Therefore, we developed a novel RT-PCR method that could reliably quantify neutrophil counts in frozen NP wash samples. mRNA transcripts of the genes encoding CD16, CD18, and CD62L were identified as neutrophil-specific in NP samples and not significantly variable in response to stimulation by heat killed bacteria, and can be used to derive an accurate assessment of neutrophil content in a sample even in the presence of epithelial cells. Using flow cytometry as a comparator, the method was validated in human NP wash samples. We conclude that this PCR-based method should prove useful for providing a quantitative estimate of neutrophil recruitment to the NP during infection and pathogenesis.

First Page

65

Last Page

70

DOI

10.1016/j.jim.2017.04.010

Volume

447

Publication Date

8-1-2017

Medical Subject Headings

Cell Count; Epithelial Cells (immunology); Flow Cytometry (methods); Humans; Nasopharynx (cytology, immunology, metabolism); Neutrophil Infiltration; Neutrophils; Polymerase Chain Reaction (methods)

PubMed ID

28454715

Link to Full Text

Share

COinS