Cathelicidin augments VDR-dependent anti-leishmanial immune response in Indian Post-Kala-Azar Dermal Leishmaniasis

Int Immunopharmacol. 2017 Sep:50:130-138. doi: 10.1016/j.intimp.2017.06.010. Epub 2017 Jul 12.

Abstract

Objectives: Indian Post kala-azar dermal leishmaniasis (PKDL) is the cutaneous aftermath of visceral leishmaniasis (VL) caused by L. donovani. Vitamin D-regulated cationic antimicrobial peptide cathelicidin (hCAP-18/LL-37) has microbicidal and immunomodulatory role against cutaneous infections, but its role in PKDL remains elusive.

Methods: Skin snips and blood-derived monocytes of PKDL patients (n=46), before (BT) and after (AT) chemotherapy, were used for this study. Serum vitamin D3 level was evaluated by ELISA. Cathelicidin and vitamin D receptor (VDR) levels were analyzed by real-time PCR and flowcytometry in PKDL patients. The mechanistic effect of cathelicidin on macrophage differentiation and anti-leishmanial activity was assessed through RNA interference techniques followed by subsequent microscopic evaluation of in vitro parasite killing and Th1/Th2 counter-regulation by ELISA/RT-PCR.

Results: Low vitamin D3 levels were accompanied with decreased expression of cathelicidin and VDR in PKDL-BT patients. Results suggested positive induction of VDR-dependent cathelicidin in PKDL macrophages by Amphotericin B treatment, which could be due to indirect effect of drug-induced IL12 upregulation. 1,25-Vitamin D3 stimulation induced cathelicidin in PKDL-BT patients through involvement of TLR2/IL-1β, but not TLR4. Cathelicidin also augmented the anti-leishmanial effect and macrophage activating potential of Amphotericin B, attributable to regulation of VDR-dependent enhancement of CD40, p-STAT-I and MHC-II expression leading to regulation of IL10/IL12 balance in PKDL-BT patient macrophages.

Conclusions: This study indicates that cathelicidin augments anti-leishmanial macrophage activating property of Amphotericin B in a TLR2/VDR dependent mechanism, and advocate the development of novel adjunct treatment modality of cathelicidin with conventional Amphotericin B in PKDL patients.

Keywords: Antimicrobials; Cathelicidin; PKDL; TLR; Visceral leishmaniasis; Vitamin D3.

MeSH terms

  • Adolescent
  • Adult
  • Amphotericin B / therapeutic use*
  • Antimicrobial Cationic Peptides / genetics
  • Antimicrobial Cationic Peptides / metabolism*
  • Cathelicidins
  • Cells, Cultured
  • Drug Therapy, Combination
  • Female
  • Humans
  • Immunity
  • Leishmania donovani / immunology*
  • Leishmaniasis, Cutaneous / immunology*
  • Macrophages / immunology*
  • Macrophages / parasitology
  • Male
  • RNA, Small Interfering / genetics
  • Receptors, Calcitriol / genetics
  • Receptors, Calcitriol / metabolism*
  • Toll-Like Receptor 2 / metabolism
  • Vitamin D / therapeutic use*
  • Young Adult

Substances

  • Antimicrobial Cationic Peptides
  • RNA, Small Interfering
  • Receptors, Calcitriol
  • TLR2 protein, human
  • Toll-Like Receptor 2
  • Vitamin D
  • Amphotericin B
  • Cathelicidins