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Caffeic acid combined with autoclaved Leishmania major boosted the protection of infected BALB/c mice by enhancing IgG2 production, IFN-γ/TGF-β and iNO synthase/arginase1 ratios, and the death of infected phagocytes

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Abstract

Background

Immunization with killed Leishmania promastigotes without adjuvant was considered as safe, but gave variable rates of protection. Taking advantage of the immuno-modulatory effect of caffeic acid (CA), a natural polyphenolic antioxidant, we investigated its potentiating effect in autoclaved Leishmania major (ALM)-induced protection of Leishmania major-infected BALB/c.

Methods

First, BALB/c mice were sensitized for 6 weeks either with CA, or ALM alone or combined with caffeic acid (ALM–CA) or Freund’s adjuvant (ALM–FA), and subsequently infected with L. major promastigotes. Second, to test the curative effect, CA was given daily for 5 weeks to susceptible mice, starting on week 4 post-infection. Sera, footpads and lymph nodes (LNs) were collected at week 9 post-infection and submitted to biochemical or histological analyses.

Results

Compared to the respective controls, our results showed that CA directly healed footpad lesions and reduced the hallmarks of cutaneous leishmaniasis including oxidative inflammation, parasite load, and phagocytes influx and infestation. In sensitized mice, the protection enhanced gradually from ALM–FA, CA, ALM to ALM–CA in parallel to decreased seric IgGt levels. In contrast to ALM–FA, the combined effect of ALM and CA increased specific isotype IgG2, and decreased IL-17 and MCP-1, and phagocyte influx, as attested by the concomitant reduction in myeloperoxidase (MPO) and α-naphthyl acetate esterase (ANAE) activities. ALM–CA shifted IFN-γ/TGF-β and iNO synthase/arginase1 (iNOS/Arg1) balances in a Th1 immune response that control efficiently cutaneous lesions and LNs hypertrophy, and reactivate the death of infected phagocytes.

Conclusions

Therefore, CA combined with ALM synergizes with L. major antigens for priming innate cells, through early polarization to optimal Th1 response that leads to IFN-γ and iNOS-dependent leishmanicidal activity of neutrophils and macrophages.

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Abbreviations

Arg1:

Arginase1

ALM:

Autoclaved Leishmania major

ANAE:

α-Naphthyl acetate esterase

CA:

Caffeic acid

DCs:

Dendritic cells

iNOS:

Inducible NO synthase

IFN-γ:

Interferon-γ

IL-17A:

Interleukin 17A

LNs:

Lymph nodes

MCP-1:

Monocyte chemoattractant protein 1

MO:

Macrophages

MPO:

Myeloperoxidase

PN:

Neutrophils

ROS:

Reactive oxygen species

TGF-β:

Tumor growth factor-β

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Acknowledgements

We are grateful to Pr Zoubir Harrat from “Laboratoire d’Eco-Epidémiologie et Génétique des Populations, Institut Pasteur, Algiers, Algeria”, for her kind gift of Leishmania major inoculum.

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  1. Laboratory of Cellular and Molecular Biology, Faculty of Biological Sciences, University of Sciences and Technology Houari Boumediene, BP 32 EL-ALIA, 16111, Bab-Ezzouar, Algiers, Algeria

    Radia Belkhelfa-Slimani & Bahia Djerdjouri

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  1. Radia Belkhelfa-Slimani
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Correspondence to Bahia Djerdjouri.

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Belkhelfa-Slimani, R., Djerdjouri, B. Caffeic acid combined with autoclaved Leishmania major boosted the protection of infected BALB/c mice by enhancing IgG2 production, IFN-γ/TGF-β and iNO synthase/arginase1 ratios, and the death of infected phagocytes. Inflammopharmacol 26, 621–634 (2018). https://doi.org/10.1007/s10787-017-0399-z

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