A clioquinol-containing Pluronic® F127 polymeric micelle system is effective in the treatment of visceral leishmaniasis in a murine model

A clioquinol (ICHQ)-containing Pluronic® F127 polymeric micelle system (ICHQ/Mic) was recently shown to be effective against Leishmania amazonensis infection in a murine model. In the present study, ICHQ/Mic was tested against L. infantum infection. BALB/c mice (n = 12 per group) were infected with L. infantum stationary promastigotes through subcutaneous injection and, 45 days after challenge, received saline or were treated via the subcutaneous route with empty micelles, ICHQ or ICHQ/Mic. In addition, animals were treated with miltefosine by the oral route, as a drug control. Half of the animals were euthanized 1 and 15 days after treatment, aiming to evaluate two endpoints after therapy, when parasitological and immunological parameters were investigated. Results showed that the treatment using miltefosine, ICHQ or ICHQ/Mic induced significantly higher anti-parasite IFN-γ, IL-12, GM-CSF, nitrite and IgG2a isotype antibody levels, which were associated with low IL-4 and IL-10 production. In addition, a higher frequency of IFN-γ and TNF-α-producing CD4+ and CD8+ T-cells was found in these animals. The parasite load was evaluated in distinct organs, and results showed that the treatment using miltefosine, ICHQ or ICHQ/Mic induced significant reductions in organic parasitism in the treated and infected mice. A comparison between the treatments suggested that ICHQ/Mic was the most effective in inducing a highly polarized Th1-type response, as well as reducing the parasite load in significant levels in the treated and infected animals. Data obtained 15 days after treatment suggested maintenance of the immunological and parasitological responses. In conclusion, ICHQ/Mic could be considered in future studies for the treatment of visceral leishmaniasis.

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Concomitant infection with Leishmania donovani and Plasmodium berghei alters clinical and immune responses in BALB/c mice

10.1101/2021.04.08.438937 ◽

2021 ◽

Author(s):

Rebeccah Moraa Ayako ◽

Joshua Muli Mutiso ◽

John Chege Macharia ◽

David Langoi ◽

Lucy Ochola

Keyword(s):

Body Weight ◽

Visceral Leishmaniasis ◽

Plasmodium Berghei ◽

Leishmania Donovani ◽

Parasite Burden ◽

Parasite Load ◽

Early Screening ◽

Immunological Parameters ◽

Geographical Regions ◽

Ifn Γ

Malaria and visceral leishmaniasis coexist in the same geographical regions. However, dual co-infection with parasites causing these diseases and their impact on public health is poorly documented. Interactions between these parasites may play a role in disease outcome. The present study set out to evaluate the clinical and immunological parameters following Leishmania donovani and Plasmodium berghei co-infection in BALB/c mice. Mice were divided into four groups; L. donovani- only, L. donovani- P. berghei , P. berghei- only and naïve. Body weight, parasite burden, total IgG, IFN-γ and IL-4 responses were determined. To determine the survival rate, four mice were used from each group. Tissues for histological analysis were taken from spleen, liver and brain. Results indicated significant differences in body weight (P<0.0001), L. donovani parasite load (P< 0.0001 ), L. donovani IgG (P< 0.0001), P. berghei parasitemia (P= 0.0222), P. berghei IgG (P= 0.002), IFN-γ (P<0.0001) and IL-4 (P<0.0001) in dual-infected mice. There was no correlation between L. donovani parasite load and IgG responses in single or dual infections, while there was a positive relationship of P. berghei parasitemia and IgG responses in the dual infection group only. Plasmodium berghei had the highest mortality rate compared to L. donovani - only and L. donovani- P. berghei infected mice groups. Histological analyses showed enlarged red and white pulps and pathological changes in the spleen, liver and brain tissues which were less pronounced in co-infected group. We conclude that L. donovani and P. berghei co-infection reduces disease severity and these changes seem to correlate with variation in serum IgG and cytokines (IFN-γ and IL-4). Therefore, the study recommends the importance of inclusion of early screening of malaria in Visceral Leishmaniasis patients in regions where malaria is co- endemic.

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Experimental Evaluation of Second-Line Oral Treatments of Visceral Leishmaniasis Caused by Leishmania infantum

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.43.1.172 ◽

1999 ◽

Vol 43 (1) ◽

pp. 172-174 ◽

Cited By ~ 34

Author(s):

Jean-Pierre Gangneux ◽

Michael Dullin ◽

Annie Sulahian ◽

Yves Jean-Francois Garin ◽

Francis Derouin

Keyword(s):

Visceral Leishmaniasis ◽

Murine Model ◽

Experimental Evaluation ◽

Leishmania Infantum ◽

Parasite Load ◽

Second Line ◽

Meglumine Antimoniate ◽

Marked Activity

ABSTRACT In a murine model of Leishmania infantum visceral leishmaniasis, metronidazole, ketoconazole, fluconazole, itraconazole, and terbinafine were less effective than antimonial agents in reducing hepatic parasite load. Ketoconazole potentiated the effect of meglumine antimoniate reference therapy through its marked activity against spleen infection.

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Treatment of murine visceral leishmaniasis using an 8-hydroxyquinoline-containing polymeric micelle system

Parasitology International ◽

10.1016/j.parint.2016.07.005 ◽

2016 ◽

Vol 65 (6) ◽

pp. 728-736 ◽

Cited By ~ 18

Author(s):

Mariana Costa Duarte ◽

Letícia Martins dos Reis Lage ◽

Daniela Pagliara Lage ◽

Vívian Tamietti Martins ◽

Ana Maria Ravena Severino Carvalho ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Polymeric Micelle ◽

Micelle System

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Antibody and cytokine levels in visceral leishmaniasis patients with varied parasitemia before, during, and after treatment in patients admitted to Arba Minch General Hospital, southern Ethiopia

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0009632 ◽

2021 ◽

Vol 15 (8) ◽

pp. e0009632

Author(s):

Dagimawie Tadesse ◽

Alemseged Abdissa ◽

Mekidim Mekonnen ◽

Tariku Belay ◽

Asrat Hailu

Keyword(s):

Visceral Leishmaniasis ◽

Leishmania Donovani ◽

Serum Levels ◽

High Serum ◽

Eastern Africa ◽

Southern Ethiopia ◽

Ifn Γ ◽

A Prospective Study ◽

Antibody Levels ◽

Tgf Β1

Background Visceral leishmaniasis is a disease caused by disseminated Leishmania donovani infection which affects almost half a million people annually. Most of the patients are reported from the Indian sub-continent, Eastern Africa and Brazil. In this study, we aimed to determine the levels of antibodies and cytokines in visceral leishmaniasis patients and to examine associations of parasitemia with the clinical states of patients. A prospective study was carried out, enrolling a total of 48 active VL patients who were evaluated before, during different time points and, three months after treatment. Serum cytokine concentrations, antibody levels, parasitemia, laboratory (hematologic and biochemical) measurements, and clinical parameters were assessed. Results Counts of WBC and platelets, and measurements of hemoglobin (Hb) increased during treatment (P ≤ 0.05). Elevated levels of circulating IL-10, IFN-γ, and TGF-β1 were measured before treatment. The observed increase in serum IL-10 remarkably declined within 7 days after the start of treatment. Anti-leishmanial antibody index (AI) was high in all VL patients irrespective of spleen aspirate parasite grade before treatment and at different times during treatment. However, a significant (P ≤ 0.05) decrease of AI was observed 120 days post-treatment. IL-2 serum levels were below the detection limit at all sampling points. Conclusions The present results suggest that IL-10, IFN-γ, and TGF-β1 can be used as markers of active visceral leishmaniasis. In addition, measuring circulating cytokines concentrations, particularly IL-10, in combination with other clinical evaluations, could be used as criteria for the cure. The observation that a high serum concentration of IFN-gamma at baseline was associated with low parasitemia deserves further investigations.

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Effect of switching glatiramer acetate formulation from 20 mg daily to 40 mg three times weekly on immune function in multiple sclerosis

Multiple Sclerosis Journal - Experimental Translational and Clinical ◽

10.1177/20552173211032323 ◽

2021 ◽

Vol 7 (3) ◽

pp. 205521732110323

Author(s):

Kouichi Ito ◽

Naoko Ito ◽

Sudhir K Yadav ◽

Shradha Suresh ◽

Yong Lin ◽

...

Keyword(s):

Inflammatory Cytokines ◽

Immune Cells ◽

Immunological Parameters ◽

Gm Csf ◽

Tnf Α ◽

Annualized Relapse Rate ◽

Ifn Γ ◽

Anti Inflammatory ◽

The Difference ◽

Pro Inflammatory Cytokines

Background Many RRMS patients who had been treated for over 20 years with GA 20 mg/ml daily (GA20) switched to 40 mg/ml three times-a-week (GA40) to reduce injection-related adverse events. Although GA40 is as effective as GA20 in reducing annualized relapse rate and MRI activity, it remains unknown how switching to GA40 from GA20 affects the development of pathogenic and regulatory immune cells. Objective To investigate the difference in immunological parameters in response to GA20 and GA40 treatments. Methods We analyzed five pro-inflammatory cytokines (IL-1β, IL-23, IL-12, IL-18, TNF-α), and three anti-inflammatory/regulatory cytokines (IL-10, IL-13, and IL-27) in serum. In addition, we analyzed six cytokines (IFN-γ, IL-17A, GM-CSF, IL-10, IL-6, and IL-27) in cultured PBMC supernatants. The development of Th1, Th17, Foxp3 Tregs, M1-like, and M2-like macrophages were examined by flow cytometry. Samples were analyzed before and 12 months post switching to GA40 or GA20. Results Pro- and anti-inflammatory cytokines were comparable between the GA40 and GA20 groups. Development of Th1, Th17, M1-like macrophages, M2-like macrophages, and Foxp3 Tregs was also comparable between the two groups. Conclusions The immunological parameters measured in RRMS patients treated with GA40 three times weekly are largely comparable to those given daily GA20 treatment.

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Genetically Modified DCs Engineered To Express PSA and/or PSMA Can Induce a Potent Immune Response Against Prostate Cancer Cells.

Blood ◽

10.1182/blood.v106.11.3075.3075 ◽

2005 ◽

Vol 106 (11) ◽

pp. 3075-3075

Author(s):

Jagdeep S. Walia ◽

Jianhui Cai ◽

Daniel H. Fowler ◽

Jeffrey A. Medin4

Keyword(s):

Prostate Cancer ◽

Immune Response ◽

T Cells ◽

Immune System ◽

Tumor Cells ◽

Murine Model ◽

Cytolytic Activity ◽

Gm Csf ◽

Tnf Α ◽

Ifn Γ

Abstract Prostate cancer (Pca) is the most frequently diagnosed cancer in American men, with an estimated 230,110 cases expected in 2004. Despite various treatment strategies for patients including androgen ablation, radical prostatectomy, radiotherapy, and chemotherapy, the incidence of recurrence remains high and there is limited impact on survival, specially for metastatic disease. Our strategy involves the use of genetically-modified dendritic cells (DCs) to induce an immune response. We have previously demonstrated in a murine model that mature DCs engineered to express prostate tumor-associated antigens (TAAs) can stimulate immune system to specifically target TAA-expressing tumor cells. In view of the heterogenous nature of Pca, we hypothesized that stimulating the immune system against two antigens simultaneously may augment the anti-tumor activity. We generated murine DCs from whole bone marrow from mice by culturing them in granulocytemonocyte colony stimulating factor (GM-CSF) and IL-4 (20ng/ml each) and later with TNF-α. During the DC development, they were transduced with a concentrated oncoretrovirus that engineers the coexpression of prostate specific antigen (PSA) and CD25 (a cell surface marker for tranduced cells) (DC-PSA) or solely the expression of prostate specific membrane antigen (DC-PSMA). Transductions of DCs resulted in 30–60% expression of the either CD25 or PSMA as checked by flowcytometry. These DCs displayed high expression of DC markers like CD11c, CD80, CD86, CD40 and MHC class II molecules. There was no change in their allostimulatory capacity as checked by mixed lymphocyte reaction. Later, mice were injected either with DC non-transduced(NT), DC-PSA, with DC-PSMA. After two immunizations at different time points, the splenocytes were collected from all the groups one week after the last immunization. These splenocytes were stimulated to become effectors and were subsequently analysed to check for IFN-γ secretion, IL-10 secretion and cytolytic assays, using the targets as syngeneic murine prostate tumor cells, RM1 engineered to express PSA and PSMA. The effectors showed high IFN-γ and high cytolytic activity low IL-10 secretion as compared to controls. Our next step will be to test the increase of the levels of IFN-γ secretion and cytolytic activity in the mice immunized with DC-PSA and DC-PSMA both as compared to DC-PSA alone and DC-PSMA alone. To show clinically feasibility of our approach, we extended our work to human cells. HuDCs were generated using human CD34+ hematopoietic cells by culturing them in GM-CSF, SCF, Flt3L and TNF-α for 12 days. During DC production, they were transduced to express PSA or PSMA using a concentrated oncoretrovirus. They were checked for DC markers and the expression of the respective TAAs i.e PSA (CD25) or PSMA. Later, these cells were co-cultured with autologous T-cells. When these immunized T cells were used as effectors against the HLA-matched prostate cancer cell lines expressing PSA and PSMA, they showed high IFN-γ secretion and Low IL-10 secretion as compared controls. Thus, we have found that human DCs can be used to sensitize T cells to show antitumor responses and we are going to test in murine model the augmentation of such antitumour response by using multiple antigen immunotherapy approach.

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Antigenicity, immunogenicity and protective efficacy of a conserved Leishmania hypothetical protein against visceral leishmaniasis

Parasitology ◽

10.1017/s0031182017001731 ◽

2017 ◽

Vol 145 (6) ◽

pp. 740-751 ◽

Cited By ~ 8

Author(s):

Daniel S. Dias ◽

Vívian T. Martins ◽

Patrícia A. F. Ribeiro ◽

Fernanda F. Ramos ◽

Daniela P. Lage ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Mononuclear Cells ◽

Protective Efficacy ◽

High Sensitivity ◽

Parasite Load ◽

Hypothetical Protein ◽

Enzyme Linked Immunosorbent Assay ◽

Peripheral Blood Mononuclear ◽

Macrophage Colony ◽

Antibody Levels

AbstractIn this study, a Leishmania hypothetical protein, LiHyS, was evaluated regarding its antigenicity, immunogenicity and protective efficacy against visceral leishmaniasis (VL). Regarding antigenicity, immunoblottings and an enzyme-linked immunosorbent assay using human and canine sera showed high sensitivity and specificity values for the recombinant protein (rLiHyS) in the diagnosis of VL. When evaluating the immunogenicity of LiHyS, which is possibly located in the parasite's flagellar pocket, proliferative assays using peripheral blood mononuclear cells from healthy subjects or VL patients showed a high proliferative index in both individuals, when compared to the results obtained using rA2 or unstimulated cultures. Later, rLiHyS/saponin was inoculated in BALB/c mice, which were then challenged with Leishmania infantum promastigotes. The vaccine induced an interferon-γ, interleukin (IL)-12 and granulocyte-macrophage colony-stimulating factor production, which was maintained after infection and which was associated with high nitrite and IgG2a antibody levels, as well as low IL-4 and IL-10 production. Significant reductions in the parasite load in liver, spleen, bone marrow and draining lymph nodes were found in these animals. In this context, the present study shows that the rLiHyS has the capacity to be evaluated as a diagnostic marker or vaccine candidate against VL.

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In vitro and in vivo antileishmanial activity of β-acetyl-digitoxin, a cardenolide of Digitalis lanata potentially useful to treat visceral leishmaniasis

Parasite ◽

10.1051/parasite/2021036 ◽

2021 ◽

Vol 28 ◽

pp. 38

Author(s):

Camila S. Freitas ◽

Daniela P. Lage ◽

João A. Oliveira-da-Silva ◽

Rafaella R. Costa ◽

Débora V.C. Mendonça ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Polymeric Micelles ◽

Direct Action ◽

Low Cost ◽

Parasite Load ◽

Antileishmanial Activity ◽

Digitalis Lanata ◽

Ifn Γ

Current treatments of visceral leishmaniasis face limitations due to drug side effects and/or high cost, along with the emergence of parasite resistance. Novel and low-cost antileishmanial agents are therefore required. We report herein the antileishmanial activity of β-acetyl-digitoxin (b-AD), a cardenolide isolated from Digitalis lanata leaves, assayed in vitro and in vivo against Leishmania infantum. Results showed direct action of b-AD against parasites, as well as efficacy for the treatment of Leishmania-infected macrophages. In vivo experiments using b-AD-containing Pluronic® F127 polymeric micelles (b-AD/Mic) to treat L. infantum-infected mice showed that this composition reduced the parasite load in distinct organs in more significant levels. It also induced the development of anti-parasite Th1-type immunity, attested by high levels of IFN-γ, IL-12, TNF-α, GM-CSF, nitrite and specific IgG2a antibodies, in addition to low IL-4 and IL-10 contents, along with higher IFN-γ-producing CD4+ and CD8+ T-cell frequency. Furthermore, low toxicity was found in the organs of the treated animals. Comparing the therapeutic effect between the treatments, b-AD/Mic was the most effective in protecting animals against infection, when compared to the other groups including miltefosine used as a drug control. Data found 15 days after treatment were similar to those obtained one day post-therapy. In conclusion, the results obtained suggest that b-AD/Mic is a promising antileishmanial agent and deserves further studies to investigate its potential to treat visceral leishmaniasis.

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Treatment of Experimental Visceral Leishmaniasis with Amphotericin B in Stable Albumin Microspheres

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.48.9.3246-3252.2004 ◽

2004 ◽

Vol 48 (9) ◽

pp. 3246-3252 ◽

Cited By ~ 43

Author(s):

J. A. Sánchez-Brunete ◽

M. A. Dea ◽

S. Rama ◽

F. Bolás ◽

J. M. Alunda ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Amphotericin B ◽

Parasite Load ◽

Bolus Administration ◽

Reference Formulation ◽

Promising Alternative ◽

Albumin Microspheres ◽

New Formulation ◽

Antibody Levels ◽

Higher Doses

ABSTRACT Hydrophilic albumin microspheres are proposed as a new delivery system for amphotericin B (AMB; AMB microspheres). The acute toxicity of AMB microspheres was lower than that of the AMB-deoxycholate (AMB-Doc) reference formulation in hamsters. Lethal doses in healthy and infected animals were improved at least eight times. Intravenous bolus administration of doses of AMB microspheres up to 40 mg/kg of body weight did not produce acute symptoms of toxicity. The efficacy of this new formulation was tested against Leishmania infantum-infected hamsters at doses of 2, 10, 20, and 40 mg/kg. With the 2-mg/kg dose, the activity of AMB, as assessed through the parasite load reductions in the liver and spleen and the evolution of antibody levels, was also improved (P < 0.05) by use of the AMB microsphere system. At the higher doses of 10, 20, and 40 mg/kg, reductions in parasite levels of more than 99% were achieved in the liver and spleen after the administration of AMB microspheres. A pharmacokinetic study was performed to study the serum, liver, and spleen AMB concentrations after administration of AMB microspheres and the reference formulation. Interestingly, a significant accumulation of AMB in the spleen and liver was observed after AMB microsphere administration. Our results suggest that this new formulation is a promising alternative to the conventional AMB-Doc formulation for the treatment of visceral leishmaniasis.

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Absence of Bim sensitizes mice to experimental Trypanosoma cruzi infection

Cell Death and Disease ◽

10.1038/s41419-021-03964-6 ◽

2021 ◽

Vol 12 (7) ◽

Author(s):

Marcela Hernández-Torres ◽

Rogério Silva do Nascimento ◽

Monica Cardozo Rebouças ◽

Alexandra Cassado ◽

Kely Catarine Matteucci ◽

...

Keyword(s):

T Cells ◽

Trypanosoma Cruzi ◽

Chagas Disease ◽

Parasite Load ◽

Peritoneal Macrophages ◽

T Cell Response ◽

No Production ◽

Similar Reduction ◽

Ifn Γ

AbstractChagas disease is a life-threatening disorder caused by the protozoan parasite Trypanosoma cruzi. Parasite-specific antibodies, CD8+ T cells, as well as IFN-γ and nitric oxide (NO) are key elements of the adaptive and innate immunity against the extracellular and intracellular forms of the parasite. Bim is a potent pro-apoptotic member of the Bcl-2 family implicated in different aspects of the immune regulation, such as negative selection of self-reactive thymocytes and elimination of antigen-specific T cells at the end of an immune response. Interestingly, the role of Bim during infections remains largely unidentified. To explore the role of Bim in Chagas disease, we infected WT, Bim+/−, Bim−/− mice with trypomastigotes forms of the Y strain of T. cruzi. Strikingly, our data revealed that Bim−/− mice exhibit a delay in the development of parasitemia followed by a deficiency in the control of parasite load in the bloodstream and a decreased survival compared to WT and Bim+/− mice. At the peak of parasitemia, peritoneal macrophages of Bim−/− mice exhibit decreased NO production, which correlated with a decrease in the pro-inflammatory Small Peritoneal Macrophage (SPM) subset. A similar reduction in NO secretion, as well as in the pro-inflammatory cytokines IFN-γ and IL-6, was also observed in Bim−/− splenocytes. Moreover, an impaired anti-T. cruzi CD8+ T-cell response was found in Bim−/− mice at this time point. Taken together, our results suggest that these alterations may contribute to the establishment of a delayed yet enlarged parasitic load observed at day 9 after infection of Bim−/− mice and place Bim as an important protein in the control of T. cruzi infections.

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