University of Tasmania
Browse

File(s) under permanent embargo

Effect of epithelial cell type on in vitro invasion of non-typeable Haemophilus influenzae

journal contribution
posted on 2023-05-18, 23:04 authored by Neeraj SinghNeeraj Singh, Kunde, DA, Stephen TristramStephen Tristram
Non-typeable Haemophilus influenzae (NTHi) have been shown to have variable ability for in vitro invasion with a range of epithelial cells, and increased invasion of BEAS-2B cells has been associated with altered penicillin binding protein3 (PBP3), which is concerning as these strains are increasing worldwide. The aim of the study was to investigate the effect of respiratory cell type and the presence of altered PBP3 on the in vitro invasion of NTHi. A collection of 16 clinical NTHi isolates was established, 7 had normal PBP3, and 9 had altered PBP3 as defined by an N526K substitution. The isolates were tested for invasion of BEAS-2B, NHBE, A549 and NCI-H292 respiratory epithelial cells in vitro using a gentamicin survival assay, with invasion measured as the percentage of intracellular organisms relative to the initial inoculum. The overall median invasion for the 16 NTHi isolates for cell types BEAS-2B, NHBE, A549 and NCI-H292 cells were 3.17, 2.31, 0.11 and 1.52 respectively. The differences were statistically significant for BEAS-2B compared to A549 (P = 0.015) and A549 compared to NCI-H292 (P = 0.015), and there were also very marked differences in invasion for some individual isolates depending on the cell type used. There was a consistent bias for invasion of isolates with normal versus abnormal PBP3: and this was statistically significant for BEAS-2B (0.07 to 9.90, P = 0.031) and A549 cells (0.02 to 1.68, P = 0.037). These results show that NTHi invasion of respiratory epithelial cells in vitro is both strain dependant and influenced significantly by the cell line used, and that the association between altered PBP3 and increased invasion is conserved across multiple cell lines.

History

Publication title

Journal of Microbiological Methods

Volume

129

Pagination

66-69

ISSN

0167-7012

Department/School

School of Health Sciences

Publisher

Elsevier Science Bv

Place of publication

Netherlands

Rights statement

Copyright 2016 Elsevier B.V

Repository Status

  • Restricted

Socio-economic Objectives

Diagnosis of human diseases and conditions

Usage metrics

    University Of Tasmania

    Exports

    RefWorks
    BibTeX
    Ref. manager
    Endnote
    DataCite
    NLM
    DC