Healing Of Cutaneous Leishmaniasis Research Articles

Consolidation of a Molecular Signature of Healing in Cutaneous Leishmaniasis Is Achieved during the First 10 Days of Treatment.

The immune response is central to the pathogenesis of cutaneous leishmaniasis (CL). However, most of our current understanding of the immune response in human CL derives from the analysis of systemic responses, which only partially reflect what occurs in the skin. In this study, we characterized the transcriptional dynamics of skin lesions during the course of treatment of CL patients and identified gene signatures and pathways associated with healing and nonhealing responses. We performed a comparative transcriptome profiling of serial skin lesion biopsies obtained before, in the middle, and at the end of treatment of CL patients (eight who were cured and eight with treatment failure). Lesion transcriptomes from patients who healed revealed recovery of the stratum corneum, suppression of the T cell-mediated inflammatory response, and damping of neutrophil activation, as early as 10 d after initiation of treatment. These transcriptional programs of healing were consolidated before lesion re-epithelization. In stark contrast, downregulation of genes involved in keratinization was observed throughout treatment in patients who did not heal, indicating that in addition to uncontrolled inflammation, treatment failure of CL is mediated by impaired mechanisms of wound healing. This work provides insights into the factors that contribute to the effective resolution of skin lesions caused by Leishmania (Viannia) species, sheds light on the consolidation of transcriptional programs of healing and nonhealing responses before the clinically apparent resolution of skin lesions, and identifies inflammatory and wound healing targets for host-directed therapies for CL.

Staphylococcus aureuspromotes increase IL-1 mediated immunopathology and delayed healing in cutaneous leishmaniasis

Abstract The chronic infection by Leishmania braziliensis causes a skin barrier (ulcer) breach and is susceptible to secondary bacterial infections. The development of the ulcer is associated with a dysregulation of the skin microbiome, which we hypothesized impacts host gene expression, exacerbates the inflammatory response, and prolongs the resolution of the disease. We conducted an integrative multi-omics meta-analysis of the skin microbiome, host gene expression, and clinical metadata of patients. We found that a high bacterial burden was associated with an increased inflammatory response and delayed healing. 16S-seq analysis of lesion swabs revealed that while patients exhibit several distinct skin microbiomes, a microbiome dominated by Staphylococcus is the most common. Furthermore, the patients with a dominant lesional Staphylococcus microbiome were delayed in healing compared to those with a heterogeneous microbiome. Since we found that S. aureus was a common isolate from the ulcers, we carried out dual RNA-seq analysis on the lesion biopsies to quantify S. aureus transcriptional abundances and associated them with host gene expression. High S. aureus transcript abundance was associated with delayed healing and increased expression of IL-1α and IL-1β. These cytokines were critical for modulating disease outcome since pathology was reduced when we neutralized either in L. braziliensis-infected mice colonized with S. aureus. These results indicate that an altered skin microbiome develops in cutaneous leishmaniasis, influencing responsiveness to anti-parasitic drug treatment. They suggest that host-directed therapies designed to reduce the bacterial burden in leishmanial lesions may be therapeutic. NIH (AI143790)

Evaluation of LL 37 Lipoprotein as Innate Immunity Marker among Sudanese Patients Cutaneous Leishmania

Background: The leishmaniasis is a group of diseases with a broad range of clinical manifestations caused by several species of parasites belonging to the genus Leishmania. LL-37/hCAP18, the only cathelicidin in human, is expressed as an 18-kDa preproprotein. The most prominent function of cathelicidins is their ability to inhibit propagation of a diverse range of microorganisms, which occurs at a micromolar range.
 Aim: The study was aimed to evaluate the LL37 plasma level in Leishmania Sudanese patients.
 Materials and Methods: In a case-control study, three hundred subjects were enrolled (200 as case and 100 controls). Five ml. of venous blood was collected in EDTA container, then plasma was obtained and stored frozen at -80ºC. LL 37 was estimated using competitive ELISA. The data were analyzed using SPSS version 21.
 Results: The results revealed that 115 (57%) of Leishimania patients were male and 85 (43%) were female. Plasma LL 37 level was significantly increased in Leishmania patients (1.30±0.71) compared to the control (0.21±0.20) with (p-value 0.000).
 Conclusion: Leishmania patients had higher levels of plasma LL37, suggesting effective anti-microbial immunity process enhancing healing of cutaneous leishmaniasis.
 Keywords: Cathelicidins; LL37, Innate Immunity, Leishmania , Sudan.

Effectiveness of amiodarone in treatment of cutaneous leishmaniasis caused by Leishmania major

Development of new chemotherapeutic agents is an essential issue in the treatment and control of a disease. This study aimed to evaluate the anti-leishmanial activity of amiodarone, an antiarrhythmic class III drug, against Leishmania major, the most prevalent etiological agent of cutaneous leishmaniasis in the old world. The proliferation of promastigotes and intracellular amastigotes in the absence or presence of amiodarone was estimated, in an in vitro study. For in vivo study, five weeks after infection of BALB/c mice with L. major, when the lesions appeared at the injection site, the mice were divided into four groups (n = 6 each); treatment was conducted for 28 consecutive days with vehicle, amiodarone at 40 mg/kg orally and glucantime at 60 mg/kg intraperitoneally. Therapy with amiodarone reduced the size of lesions compared to the untreated group after 12 days. Amiodarone decreased the parasite load and inflammatory responses, particularly the macrophages containing amastigotes, and enhanced granulation tissue formation in the dermis and subcutaneous area. The Tumor necrosis factor-α and Interleukin-6 levels were significantly lower in the cell culture supernatants of the inguinal lymph node in the amiodarone treated group compared to the vehicle and untreated groups. Amiodarone significantly increased the activity of glutathione peroxidase in comparison to the vehicle and untreated groups but did not affect the plasma levels of superoxide dismutase, malondialdehyde, adiponectin, and ferric reducing ability of plasma. Therefore, the anti- L. major activity and immunomodulatory effects of amiodarone reduced the parasitic load and enhanced wound healing in cutaneous leishmaniasis in BALB/c mice. Amiodarone reduced the lesion surface area, but it did not cure it completely.

Label-free quantitative proteomic analysis reveals potential biomarkers for early healing in cutaneous leishmaniasis.

BackgroundLeishmaniasis is a parasitic disease caused by more than 20 species of the Leishmania genus. The disease is globally distributed and is endemic in 97 countries and three territories in the tropical and subtropical regions. The efficacy of the current treatments is becoming increasingly low either due to incomplete treatment or resistant parasites. Failure of treatment is frequent, and therefore, the search for early biomarkers of therapeutic response in cutaneous leishmaniasis (CL) is urgently needed.ObjectiveThe aim of this study was to compare the proteomic profiles in patients with CL before and after 7 days of treatment and identify early biomarkers of curative response.MethodsFour patients with a parasitological diagnosis of leishmaniasis with confirmation of species by PCR-RFLP were recruited. All patients had a single lesion, and a protein from the middle of the ulcer was quantified by liquid chromatography and mass spectrometry.ResultsA total of 12 proteins showed differential expression in the comparative LC-electrospray ionization MS/MS (LC-ESI-MS/MS) triplicate analysis. Seven of them were up-regulated and five of them were down-regulated. Calcium binding proteins A2, A8, and A9 and hemoglobin subunits alpha-2 and delta showed high correlation with epidermis development and immune response.ConclusionWe identified changes in the profiles of proteins that had a positive therapeutic response to the treatment. The proteins identified with differential expression are related to the reduction of inflammation and increased tissue repair. These proteins can be useful as biomarkers for early monitoring of therapeutic response in CL.

Wound healing in cutaneous leishmaniasis: A double edged sword of IL-10 and TGF-β.

Immune responses have a crucial role during the wound healing process in cutaneous leishmaniasis (CL). However, there are several paradoxes in immunity against CL. On the one hand, regulatory cytokines interleukin (IL)-10 and transforming growth factor beta (TGF-β) increase susceptibility to CL through suppression of several proinflammatory cytokines that require for defense against CL. On the other hand, these cytokines play a pivotal role in the acceleration of wound healing process. This review discusses about the dual role of IL-10 and TGF-β during the wound healing process and immunity against CL to offer a new insight about wound healing in CL.

Resolving lesions in human cutaneous leishmaniasis predominantly harbour chemokine receptor CXCR3-positive T helper 1/T cytotoxic type 1 cells

Cutaneous leishmaniasis (CL) is an epidemic disease affecting millions of individuals worldwide. Treatment options have several side-effects and a vaccine does not exist at present. To translate information about protection against CL from mice to man, we studied the local immune response in CL skin biopsies and correlated these findings with clinical information. The frequency of inflammatory cells was determined in skin biopsies of 20 patients diagnosed with CL using immunohistochemistry. In addition, the nature of the resulting adaptive immune response was assessed by (double) immunostaining against CD4 and chemokine receptors CXCR3 (T helper 1, Th1)/CCR4 (Th2). All lesions contained CD4+ and CD8+ T cells, B cells and CD68+ macrophages. CD1a+ epidermal Langerhans cells were absent above the centre of the lesions, but normally distributed in the surrounding tissue. Mast cell and CD56+ natural killer cell numbers were not affected. Interestingly, CCR4+ Th2 cells were not detected in any of the 20 samples. In contrast, the number of infiltrating CXCR3+ cells was high and the majority of these were CD4+ or CD8+ indicating that they represent interferon-gamma-producing Th1/T cytotoxic type 1 (Tc1) cells. Finally, these findings did not correlate with clinical information about the country where the infection was acquired, or age or sex of the patients. However, lesions that had already persisted for more than 6 months contained fewer CXCR3+ CD4 and CD8 T cells than those that had persisted for less than 6 months. Our data on the inflammatory infiltrate of human CL lesions underline the relevance of findings obtained in experimental models. Both Th1 and Tc1 cells appear to be critical for healing in CL in mouse and man.

Production of interferon gamma in cultures of whole blood obtained in the course of and after healing of cutaneous leishmaniasis

Whole-blood cultures from individuals with cutaneous leishmaniasis due to Leishmania major, tested under field conditions, showed a good correlation between the production of interferon-gamma and total blood lymphocyte proliferation (TLP), assayed by response to specific leishmania antigen. A group of patients, re-tested on several occasions, converted to positive in both parameters of cell mediated immunity (CMI) at the same time. It is suggested that the TLP assay and the production of interferon-gamma detect similar CMI activity in human cutaneous leishmaniasis. The measurement of interferon-gamma production has the advantage over the TLP assay that the supernatants of the cultures can be frozen and stored for prolonged periods before the assay is performed. For this reason, this could present a suitable method for field studies.