Identification and Characterization of the miRNAs and Cytokines in Response to Leishmania infantum Infection with Different Response to Treatment

Abstract

Purpose: Due to the complexity of cytokine and microRNA function in progression and/or suppression of an infection, in this study, we examined miR-3473f, miR-2128, miR-6994-5p, miR-7093-3p, miR-5128, miR-574-5p, miR-7235, IL-2, IL-4, IL-5, IL-10 and IL-13 in patients with VL caused by Leishmania infantum in an in vivo study.

Methods: Sampling was carried out from patient with leishmaniasis and with different responses to treatment during March 2016-January 2020. DNA was extracted and purified using QIAamp Kit. The L. infantum were cultured in DMEM medium and protein content was determined by the Micro BCA Protein Assay Kit. Cytokines were evaluated using a MILLIPLEX MAP Mouse Cytokine/Chemokine Panel I kit. The relative expression of miRNAs was measured in duplicate using automatic thermocycler ABI Prism 7500 sequence detection system (Applied Bio-systems) using the TaqMan MicroRNA Assay kit.

Results: The real-time PCR assay revealed that miR-2128, miR-6994-5p, miR-7093-3p, miR-5128, miR-574-5p and miR-7235 were down-regulated and miR-3473 were up-regulated in patients with semi-resistance and resistance parasite strain (P < 0.05). In the current work, cytokine patterns in patients who were slow-to-clear or unable-to-clear L. infantum infection during drug treatment were seen to have decreased protective Th1 cytokines (IL-2, IL-12, TNF-α, and IFN-ɤ, P < 0.001) and increased Th2 cytokines (IL-5, IL-10, and IL-13, P < 0.001). No association was seen with IL-4 in patients with different treatment outcomes.

Conclusion: Overall, the results of a recent study have shown that cytokines and microRNAs can play a key role in response to treatment, and more comprehensive studies are needed to support this hypothesis.

Keywords: Cytokine; L. infantum; Leishmaniasis; miRNAs.