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I n Vitro Anti-inflammatory and Immunomodulatory Effects of Ciprofloxacin or Azithromycin in Staphylococcus aureus-Stimulated Murine Macrophages are Beneficial in the Presence of Cytochalasin D

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ABSTRACT

We hypothesized that if internalization of Staphylococcus aureus could be blocked by using cytochalasin D (an inhibitor of phagocytosis and phagolysosome fusion), then the intracellular entry and survival of the pathogen in host’s phagocytic cells recruited to the inflammatory site can be restricted. At the same time, if we use antimicrobial agents (e.g., ciprofloxacin and azithromycin) having potent intracellular and extracellular microbicidal activity against the bacterium that have not entered into the phagosome and remains adhered to the phagocytic cell membrane, then they can be eradicated from the site of infection without compromising the host cell. To validate this, role of ciprofloxacin (CIP) and azithromycin (AZM) in eliminating S. aureus by suppressing the phagocytic activity of macrophages with cytochalasin D before infection was investigated. CIP and AZM were used either alone or in combination with cytochalasin D. Supernatant and lysate obtained from the culture of macrophages were used for quantification of reactive oxygen species, lysozymes, antioxidant enzymes, and cytokines produced. Azithromycin was better than ciprofloxacin in combination with cytochalasin D for eradicating S. aureus and regulating cytokine release. Further studies are required for ensuring proper delivery of this combination at the site of infection.

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Abbreviations

AZM:

Azithromycin

CIP:

Ciprofloxacin

CPCSEA:

Committee for the Purpose of Control and Supervision of Experiments on Animals

DMSO:

Dimethyl sulfoxide

DTNB:

5,5′-Dithiobis-2-nitrobenzoic acid

ELISA:

Enzyme-inked immunosorbent assay

FBS:

Fetal bovine serum

FIC:

Fractional inhibitory concentration

HBSS:

Hank’s balanced salt solution

IFN-γ:

Interferon-gamma

IL:

Interleukin

IU:

International unit

MHB:

Mueller-Hinton broth

MRSA:

Methicillin-resistant S. aureus

NADPH:

Nicotinamide adenine dinucleotide phosphatase

NaOH:

Sodium hydroxide

NCCLS:

National Committee for Clinical Laboratory Standards

NF-κB:

Nuclear factor-kappa beta

OD:

Optical density

PMN:

Polymorphonuclear neutrophil

RPMI:

Roswell Park Memorial Institute

SOD:

Superoxide dismutase

TCA:

Trichloroacetic acid

TLR:

Toll-like receptor

TNF-α:

Tumor necrosis factor-alpha

Tris–EDTA–HCL:

Tris-ethylenediamine hydrochloric acid

TSST:

Toxic shock syndrome toxin

VRSA:

Vancomycin-resistant S. aureus strains

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ACKNOWLEDGMENTS

This work was supported by the Department of Biotechnology (DBT), Government of West Bengal, Calcutta, West Bengal [Grant Number: vide sanction order 786-BT/Estt/RD-4/13 date: 30.10.2013 to BB] for funding this project. The author (Biswadev Bishayi) is indebted to the Department of Science and Technology, Government of India for providing with the instruments procured under the DST-PURSE program to the Department of Physiology, University of Calcutta.

CONFLICT OF INTEREST

For the manuscript entitled “In vitro anti-inflammatory and immunomodulatory effects of ciprofloxacin or azithromycin in Staphylococcus aureus stimulated murine macrophages are beneficial in the presence of cytochalasin D” by Somrita Dey et al., the authors declared that they have no conflict of interest for this manuscript toward submission in Inflammation. The authors also state that they do not have a direct financial relation with the commercial identities mentioned in this manuscript that might lead to a conflict of interest for any of the authors.

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  1. Department of Physiology, Immunology Laboratory, University Colleges of Science and Technology, University of Calcutta, 92 APC Road, Calcutta, 700009, West Bengal, India

    Somrita Dey, Arnab Majhi, Sayantika Mahanti, Ipsita Dey & Biswadev Bishayi

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Dey, S., Majhi, A., Mahanti, S. et al. I n Vitro Anti-inflammatory and Immunomodulatory Effects of Ciprofloxacin or Azithromycin in Staphylococcus aureus-Stimulated Murine Macrophages are Beneficial in the Presence of Cytochalasin D. Inflammation 38, 1050–1069 (2015). https://doi.org/10.1007/s10753-014-0070-4

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