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The monocytosis during human leptospirosis is associated with modest immune cell activation states

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Abstract

Leptospirosis is a life-threatening zoonotic disease and it has been hypothesized that the innate immune system fails to control the infection through ill-characterized mechanisms. The aim of this observational study was to better evaluate the activation processes of monocytes at the early stage of the disease. Blood samples were taken from healthy donors (n = 37) and patients hospitalized for either non-severe (n = 25) or severe (n = 32) leptospirosis. Monocyte cell counts and phenotypes were assessed by flow cytometry. We analysed the expression of several cell activation markers: CD14, CD16, HLA-DR, CD69, TLR2, TLR4, CD11b and CD11c. Although monocyte values at admittance were not significantly different from controls, patients experienced significant monocytosis at 1.33 × 109/L (p < 0.0001 compared to controls: 0.56 × 109/L) during their hospital stay. This monocytosis observed during hospital stay was correlated to several surrogate markers of organ injury. Non-classical (CD14−CD16+) and intermediate (CD14+CD16+) monocyte subsets increased compared to controls (p < 0.05). Accordingly, classical monocyte subset (CD14+CD16−) showed decreased percentages (p < 0.0001). Levels of several cell surface activation molecules were decreased: HLA-DR involved in MHC class II antigen presentation, integrins CD11b and CD11c implicated in phagocytosis and cell recruitment (p < 0.0001). None of these parameters had a prognostic value. Results from this study showed that during acute human leptospirosis, patients experienced monocytosis with a switch toward an inflammation-related phenotype contrasted by low expression levels of markers implicated in monocyte function.

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Data availability

The datasets generated during and/or analysed during the current study are available in the FlowRepository public database, URL: https://flowrepository.org/id/RvFrfom7jwL1SHc4nTFg2epdGcDpHc3qwtsGSOPoDm6ofWkmfjfQTm22c2UcA1At.

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Acknowledgements

The authors thank the practitioners implicated in the patients’ care as well as the members of the microbiology laboratory of St Denis, La Réunion, for their help in patients’ recruitment. We also thank Alexander Greenshields for critical appraisal of the manuscript.

Funding

The authors work is funded by the research unit PIMIT (processus infectieux en milieu insulaire tropical) and the CHU (centre hospitalo-universaire) de la Réunion. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Author notes

  1. Loic Raffray and Claude Giry contributed equally.

Authors and Affiliations

  1. Université de La Réunion, CNRS 9192, INSERM U1187, IRD 249, UMR PIMIT, CHU de La Réunion, Unité Mixte Processus Infectieux en Milieu Insulaire Tropical (PIMIT), Plateforme Technologique CYROI, Sainte-Clotilde, La Réunion, France

    Loic Raffray, Marie-Christine Jaffar-Bandjee & Philippe Gasque

  2. Internal Medicine Unit, CHU La Réunion site Félix Guyon, Allée des Topazes, St Denis, La Réunion, France

    Loic Raffray & Anne Gerber

  3. CNR arboviroses, CHU La Réunion site Félix Guyon, Allée des Topazes, St Denis, La Réunion, France

    Marie-Christine Jaffar-Bandjee

  4. Microbiology/Virology Laboratory, CHU La Réunion site Félix Guyon, Allée des Topazes, St Denis, La Réunion, France

    Claude Giry & Marie-Christine Jaffar-Bandjee

  5. Intensive Care Unit, CHU La Réunion site Félix Guyon, Allée des Topazes, St Denis, La Réunion, France

    David Vandroux

  6. Internal Medicine Unit, GHER Hospital, St Benoit, La Réunion, France

    Stéphanie Fayeulle

  7. Infectious and Tropical Diseases Unit, CHU La Réunion site Félix Guyon, Allée des Topazes, St Denis, La Réunion, France

    Marie-Pierre Moiton

  8. Biology laboratory, Immunology sector, LICE-OI, CHU La Réunion site Félix Guyon, Allée des Topazes, St Denis, La Réunion, France

    Philippe Gasque

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  1. Loic Raffray
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Correspondence to Loic Raffray.

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Ethical approval

This study was approved by the local human ethic committee of tertiary teaching Hospital ‘CHU de La Réunion’ (protocol number R15018) and conducted according to the principles expressed in the Declaration of Helsinki. Informed consent was obtained from all individual participants included in the study.

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Edited by: C.Bogdan.

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Supplementary Fig. 1: Evolution of monocyte cell counts during hospitalization and convalescent phase for 10 leptospirosis patients. Caption

: Paired values of monocytes for leptospirosis cases during hospitalization (admittance and maximal value) and 1 month after discharge (M1). Comparisons with non-parametric Wilcoxon paired test. ** indicates P value inferior to 0.01, respectively. (TIFF 315 KB)

Supplementary Fig. 2: Higher

bacteraemia in severe leptospirosis cases. Caption: Plasmatic Leptospira load is established from the 50 patients with positive RT-qPCR in blood and using the log-transformed standard curve. Horizontal bars indicate the median and IQR ranges. Comparison with non-parametric Mann-Whitney test. ** indicates P value inferior to 0.01. (TIFF 289 KB)

Supplementary Fig. 3: Higher levels of cell surface expression of CD69 on monocytes in severe cases of leptospirosis Caption

: comparison of selected cell surface markers among healthy donors and leptospirosis patients according to disease severity. Horizontal bars indicate the median and IQR ranges for mean fluorescence intensity (MFI). Comparison with non-parametric Mann-Whitney test. **, **** indicate P values inferior to 0.01 and 0.0001, respectively. (TIFF 667 KB)

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Raffray, L., Giry, C., Vandroux, D. et al. The monocytosis during human leptospirosis is associated with modest immune cell activation states. Med Microbiol Immunol 208, 667–678 (2019). https://doi.org/10.1007/s00430-018-0575-9

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