Antimony resistant Leishmania donovani but not sensitive ones drives greater frequency of potent T-regulatory cells upon interaction with human PBMCs: role of IL-10 and TGF-β in early immune response

PLoS Negl Trop Dis. 2014 Jul 17;8(7):e2995. doi: 10.1371/journal.pntd.0002995. eCollection 2014 Jul.

Abstract

In India the sand fly, Phlebotomus argentipes, transmitted parasitic disease termed kala-azar is caused by Leishmania donovani (LD) in humans. These immune-evading parasites have increasingly developed resistance to the drug sodium antimony gluconate in endemic regions. Lack of early diagnosis methods for the disease limits the information available regarding the early interactions of this parasite with either human tissues or cell lineages. We reasoned that peripheral blood mononuclear cells (PBMCs) from healthy human beings could help compare some of their immune signatures once they were exposed for up to 8 days, to either pentavalent antimony sensitive (Sb(S)-LD) or resistant (Sb(R)-LD) Leishmania donovani isolates. At day 2, PBMC cultures exposed to Sb(S)-LD and Sb(R)-LD stationary phase promastigotes had four and seven fold higher frequency of IL-10 secreting monocyte-macrophage respectively, compared to cultures unexposed to parasites. Contrasting with the CD4(+)CD25-CD127- type-1 T-regulatory (Tr1) cell population that displayed similar features whatever the culture conditions, there was a pronounced increase in the IL-10 producing CD4(+)CD25(+)CD127low/- inducible T-regulatory cells (iTregs) in the PBMC cultures sampled at day 8 post addition of Sb(R)-LD. Sorted iTregs from different cultures on day 8 were added to anti-CD3/CD28 induced naïve PBMCs to assess their suppressive ability. We observed that iTregs from Sb(R)-LD exposed PBMCs had more pronounced suppressive ability compared to Sb(S)-LD counterpart on a per cell basis and is dependent on both IL-10 and TGF-β, whereas IL-10 being the major factor contributing to the suppressive ability of iTregs sorted from PBMC cultures exposed to Sb(S)-LD. Of note, iTreg population frequency value remained at the basal level after addition of genetically modified Sb(R)-LD lacking unique terminal sugar in surface glycan. Even with limitations of this artificial in vitro model of L. donovani-human PBMC interactions, the present findings suggest that Sb(R)-LD have higher immunomodulatory capacity which may favour aggressive pathology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antimony / pharmacology*
  • Antiprotozoal Agents / pharmacology*
  • Cells, Cultured
  • Humans
  • Interleukin-10 / immunology*
  • Leishmania donovani / immunology*
  • Leukocytes, Mononuclear / immunology*
  • T-Lymphocytes, Regulatory / immunology*
  • Transforming Growth Factor beta / immunology*

Substances

  • Antiprotozoal Agents
  • IL10 protein, human
  • Transforming Growth Factor beta
  • Interleukin-10
  • Antimony

Grants and funding

This research was supported by the European Commission-funded Kaladrug-R Project (Health-F3-2008-222895) and the Council of Scientific and Industrial Research (grant no. BSC-0114 and BSC-0120), New Delhi. Fellowships from the CSIR, Government of India was awarded to RG, SD, and JG. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.