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Rapid onset of ICAM-1 expression is a marker of effective macrophages activation during infection of Francisella tularensis LVS in vitro

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Abstract

Francisella tularensis is capable to modulate immunobiological activities of the host cells. We focused on the expression of ICAM-1 (CD54) on J774.2 mouse macrophage cell line infected by F. tularensis live vaccine strain (LVS) in vitro as a putative marker of subsequent elimination of infection. J774.2 cell line cells were infected by F. tularensis LVS strain (multiplicity of infection, 1:100). Cell cultures were stimulated either 3 h before infection or 3 h after infection by either lipopolysaccharide (LPS) or interferon γ (IFN-γ). The expression of ICAM-1 was determined by flow cytometry 6 h after infection. The intensity of ICAM-1 expression after 6 h of J774.2 macrophage cells infection by F. tularensis is very sensitive indicator of the effective macrophages stimulation resulting in the elimination of F. tularensis infection. The mean fluorescence intensity MFI = 49.8 is set-up by our experiments as a reliable threshold of the effective elimination of F. tularensis experimental infection with 83.3% sensitivity and 96.7% specificity, respectively. Simultaneous stimulation of J774.2 macrophage cells by LPS and IFN-γ was essential to elicit the elimination of F. tularensis infection. The ICAM-1 expression determined by flow cytometry can be considered to be highly sensitive and specific approach to predict elimination of F. tularensis infection by J774.2 macrophages.

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Abbreviations

ATCC:

American Type Culture Collection

CDC:

Centre for Disease Control

CFU:

Colony forming units

ETCC:

European Type Culture Collection

FITC:

Fluorescein isothiocyanate

F. tularensis :

Francisella tularensis

HSD:

Honestly significant difference (post hoc test)

ICAM:

Intercellular adhesion molecule

IL:

Interleukin

IFN-γ:

Interferon γ

LPS:

Lipopolysaccharide

LVS:

Live vaccine strain

MAPK:

Mitogen-activated protein kinase

MEM:

Modified Eagle’s medium

MFI:

mean fluorescence index

MOI:

Multiplicity of infection

NFκB:

Nuclear factor κB

PBS:

Phosphate-buffered saline

ROC:

Receiver operating characteristics curve

AUROC:

Area under ROC curve)

TLR:

Toll-like receptor

TNF-α:

Tumor necrosis factor α

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Acknowledgment

The study was supported by grant no. LN00A033 and, in part, also by project no. MSM0021620812 of the Ministry of Education, Youth and Sport, Czech Republic.

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Authors and Affiliations

  1. Department of Clinical Immunology, Faculty of Medicine and University Hospital in Hradec Kralove, Charles University Prague, Sokolska 581, 500 05, Hradec Kralove, Czech Republic

    J. Novosad, M. Holicka, M. Novosadova, J. Krejsek & I. Krcmova

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  1. J. Novosad
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  2. M. Holicka
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  3. M. Novosadova
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  4. J. Krejsek
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  5. I. Krcmova
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Correspondence to J. Krejsek.

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Novosad, J., Holicka, M., Novosadova, M. et al. Rapid onset of ICAM-1 expression is a marker of effective macrophages activation during infection of Francisella tularensis LVS in vitro. Folia Microbiol 56, 149–154 (2011). https://doi.org/10.1007/s12223-011-0028-y

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  • DOI: https://doi.org/10.1007/s12223-011-0028-y

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