Cytokines and microbicidal molecules regulated by IL-32 in THP-1-derived human macrophages infected with New World Leishmania species

Abstract

BackgroundInterleukin-32 (IL-32) is expressed in lesions of patients with American Tegumentary Leishmaniasis (ATL), but its precise role in the disease remains unknown.Methodology/Principal findingsIn the present study, silencing and overexpression of IL-32 was performed in THP-1-derived macrophages infected with Leishmania (Viannia) braziliensis or L. (Leishmania) amazonensis to investigate the role of IL-32 in infection. We report that Leishmania species induces IL-32γ, and show that intracellular IL-32γ protein production is dependent on endogenous TNFα. Silencing or overexpression of IL-32 demonstrated that this cytokine is closely related to TNFα and IL-8. Remarkably, the infection index was augmented in the absence of IL-32 and decreased in cells overexpressing this cytokine. Mechanistically, these effects can be explained by nitric oxide cathelicidin and β-defensin 2 production regulated by IL-32.ConclusionsThus, endogenous IL-32 is a crucial cytokine involved in the host defense against Leishmania parasites.

Genetic variation in Interleukin-32 influence the immune response against New World Leishmania species and susceptibility to American Tegumentary Leishmaniasis

“…cDNA was made using the iScript kit (Bio-Rad). Primer pairs for the IL32 isoforms are described elsewhere [36]. IL1B, IL1A, IL22, IL-17 and IFNG primers sequences were designed specifically for each transcript using Primer Express 3.0 software (DeNovo Software, Glendale, CA, USA).…”

“…We previously described the expression of IL-32 in epithelial, endothelial and mononuclear cells in lesions from patients with LCL and ML [35]. In fact, IL-32 has also been implicated as a key player in controlling parasite load in infections caused by L. braziliensis or L. amazonensis in both the IL-32γ-transgenic mouse model and human cells [36,37]. To date, it is not precisely known how IL-32 isoforms contribute to host defense against Leishmania infections.…”

Interleukin-32 upregulates the expression of ABCA1 and ABCG1 resulting in reduced intracellular lipid concentrations in primary human hepatocytes

“…2.5 Â 10 6 cells/800 mL were electroporated with Amaxa Nucleofactor technology (Lonza, Basel), according with the protocol described in [35]. For IL-32 knockdown, 1 mg of ON-TARGETplus SMARTpool siRNA per transfection was used or 1 mg of ON-TARGETplus SMARTpool control siRNA (Dharmacon Inc.), sequences are described in [36]. For IL-32 overexpression, 0.5 mg of pCDNA3 plasmid expressing human IL-32g of eGFP was used as control.…”

Interferon-Beta Treatment Differentially Alters TLR2 and TLR4-Dependent Cytokine Production in Multiple Sclerosis Patients

“…The mRNA was isolated by the TRIzol (Invitrogen, Carlsbad, CA, USA) method, and quantitative polymerase chain reaction (qPCR) was performed as described by Dos Santos et al [25]. The mRNA analysis was done with the 2 × dCt × 1,000 method and normalized using the constitutive GAPDH gene.…”